Biomarkers of Inflammation Kussmann
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Transcript of Biomarkers of Inflammation Kussmann
MARKERS AND LAB TESTS OF INFLAMMATION EVIDENCE IN FAVOUR OF TOLLE CAUSAM USE OF NATUROPATHIC MEDICINE TO REDUCE THE
INCIDENCE OF CHRONIC DISEASE
Heidi Kussmann ND FABNO
THE VALUE OF LAB MEDICINE TO HEALTH CARE
bull Objective data that enables
bull Screening for risk factors
bull Accurate and early diagnosis
bull Determination of disease severity
bull Recoverysurvivalprognostic value
bull Treatment selection
bull Evaluation of adverse eventsoutcomes
bull Chronic diseasemedication management
Subjective frustration translation to whole systems
THE VALUE OF LAB MEDICINE TO HEALTH CARE
bull Services provided
bull Matching blood to patient
bull Public health infectious disease identification and control
bull Rapid testing for triage and emergency care
bull Assessment of quality of care in chronic diseases
bull A basis for evidence-based medicine for cost-effective testing therapeutic treatment and interventions screening and design of therapies based in pharmacogenomics or nutrigenomics
2httpswwwfuturelabmedicineorgpdfs200720status20report20laboratory_medicine_-_a_national_status_report_from_the_lewin_grouppdfaccessed Dec 15 2012
HISTORY OF LAB SCIENCES AND INFLAMMATIONIt all started whenhellip
BRIEF HISTORY OF MEDICAL DIAGNOSIS
bull Middle ages-18th century bedside medicine
bull 1794-1848 hospital medicine
bull 1848-present laboratory medicine
bull Laboratory medicine is appreciated as the seat of modern medicine
bull Early lab techniques of diagnosis
bull Observation (palpation taste vision smell hearing)
bull Late 19th century tools Microscope thermometer stethoscope
FOUNDER OF EXPERIMENTAL PHYSIOLOGY ndash GALEN (AD
131-201)bull Combined hippocrates humoral and pythagorean
theories to correspond to various combinations of drycoldhotmoist with earthairfirewater
bull Hot + moist = blood
bull Hot + dry = yellow bile
bull Cold + moist = phlegm
bull Cold + dry = black bile
bull Ancient diagnostic methods3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
The 5 cardinal signs of inflammation originally described by Celsus 2000 years ago
Source httpwwwnaturecomnrijournalv2n10fig_tabnri915_F1html accessed March 8 2013
INFLAMMATORY MARKERS AND TESTS USED IN THE
MIDDLE AGES
bullUrineStool inspection
bull Temperature
bullPulse
bullPalpitation
bullPercussion
bullDreams3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
17TH CENTURY ndash A CENTURY OF URINE
bull William Harvey ndash blood circulation
bull 32-power microscope
led to the study of cellshistology
bull Urine tasting to differentially diagnose diabetes insipidus and diabetes mellitus
bull Urinalysis became more qualitative
bull Blood transfusions IV drug injections
bull Pulse and temperature as indicators of health3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
18TH CENTURY ndash THEORETICAL ADVANCEMENT OF MEDICINE
bull Digitalis and opium discovered for medical use
bull Septicemia was highest cause of mortality in hospitals
bull Prothrombin time and plasma prothrombin time fibrinogen (ldquocoagulable lymphrdquo) was discovered as the cause of coagulation by William Hewson (1739-1774)
bull Pulse measurement refined but rejected due to skepticism for the Galenic doctrine of there being a special pulse for every disease
bull Arterial blood pressure measurement quantified by Stephen Hales (1677-1761)
bull Discovery of sediment in the urine of febrile patientsBerger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
19TH CENTURY DATA
The microscope the X-raybull Enabled several doctors to view and discuss resultsbull Team approach to diagnosis strengthened the medical
communityrsquos claim to objective judgment
Set standards for physiology deviations and classifications
Quantitative diagnosis in the hospital laboratory allowed diagnosis of TB cholera typhoid diphtheria and syphilis
Standardized testing became the norm
Hematological testing discoveries calcium salts in the coagulation of blood mast cells white blood cell classifications
3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
INFLAMMATION GOOD BAD UGLY
bull Good necessary to promote an immune response in otherwise healthy individuals
bull Bad in allergenic and auto-immunogenic unhealthy individuals
bull Ugly chronic disease and cancer are the result of long-standing chronic inflammation
Inflammation is an incendiary process that if properly detected and treated could save lives
THE START OF INFLAMMATION
bull The start of inflammation depends on the genetics of the host and that individualrsquos response
bull Oxidized pholpholipids are potent stimuli that trigger macrophage recruitment and contribute to both the initiation and progression of chronic inflammation
4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
5 Berliner JA Leitinger N and Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis J Lipid Res Suppl 50 S207ndashS212
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
THE VALUE OF LAB MEDICINE TO HEALTH CARE
bull Objective data that enables
bull Screening for risk factors
bull Accurate and early diagnosis
bull Determination of disease severity
bull Recoverysurvivalprognostic value
bull Treatment selection
bull Evaluation of adverse eventsoutcomes
bull Chronic diseasemedication management
Subjective frustration translation to whole systems
THE VALUE OF LAB MEDICINE TO HEALTH CARE
bull Services provided
bull Matching blood to patient
bull Public health infectious disease identification and control
bull Rapid testing for triage and emergency care
bull Assessment of quality of care in chronic diseases
bull A basis for evidence-based medicine for cost-effective testing therapeutic treatment and interventions screening and design of therapies based in pharmacogenomics or nutrigenomics
2httpswwwfuturelabmedicineorgpdfs200720status20report20laboratory_medicine_-_a_national_status_report_from_the_lewin_grouppdfaccessed Dec 15 2012
HISTORY OF LAB SCIENCES AND INFLAMMATIONIt all started whenhellip
BRIEF HISTORY OF MEDICAL DIAGNOSIS
bull Middle ages-18th century bedside medicine
bull 1794-1848 hospital medicine
bull 1848-present laboratory medicine
bull Laboratory medicine is appreciated as the seat of modern medicine
bull Early lab techniques of diagnosis
bull Observation (palpation taste vision smell hearing)
bull Late 19th century tools Microscope thermometer stethoscope
FOUNDER OF EXPERIMENTAL PHYSIOLOGY ndash GALEN (AD
131-201)bull Combined hippocrates humoral and pythagorean
theories to correspond to various combinations of drycoldhotmoist with earthairfirewater
bull Hot + moist = blood
bull Hot + dry = yellow bile
bull Cold + moist = phlegm
bull Cold + dry = black bile
bull Ancient diagnostic methods3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
The 5 cardinal signs of inflammation originally described by Celsus 2000 years ago
Source httpwwwnaturecomnrijournalv2n10fig_tabnri915_F1html accessed March 8 2013
INFLAMMATORY MARKERS AND TESTS USED IN THE
MIDDLE AGES
bullUrineStool inspection
bull Temperature
bullPulse
bullPalpitation
bullPercussion
bullDreams3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
17TH CENTURY ndash A CENTURY OF URINE
bull William Harvey ndash blood circulation
bull 32-power microscope
led to the study of cellshistology
bull Urine tasting to differentially diagnose diabetes insipidus and diabetes mellitus
bull Urinalysis became more qualitative
bull Blood transfusions IV drug injections
bull Pulse and temperature as indicators of health3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
18TH CENTURY ndash THEORETICAL ADVANCEMENT OF MEDICINE
bull Digitalis and opium discovered for medical use
bull Septicemia was highest cause of mortality in hospitals
bull Prothrombin time and plasma prothrombin time fibrinogen (ldquocoagulable lymphrdquo) was discovered as the cause of coagulation by William Hewson (1739-1774)
bull Pulse measurement refined but rejected due to skepticism for the Galenic doctrine of there being a special pulse for every disease
bull Arterial blood pressure measurement quantified by Stephen Hales (1677-1761)
bull Discovery of sediment in the urine of febrile patientsBerger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
19TH CENTURY DATA
The microscope the X-raybull Enabled several doctors to view and discuss resultsbull Team approach to diagnosis strengthened the medical
communityrsquos claim to objective judgment
Set standards for physiology deviations and classifications
Quantitative diagnosis in the hospital laboratory allowed diagnosis of TB cholera typhoid diphtheria and syphilis
Standardized testing became the norm
Hematological testing discoveries calcium salts in the coagulation of blood mast cells white blood cell classifications
3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
INFLAMMATION GOOD BAD UGLY
bull Good necessary to promote an immune response in otherwise healthy individuals
bull Bad in allergenic and auto-immunogenic unhealthy individuals
bull Ugly chronic disease and cancer are the result of long-standing chronic inflammation
Inflammation is an incendiary process that if properly detected and treated could save lives
THE START OF INFLAMMATION
bull The start of inflammation depends on the genetics of the host and that individualrsquos response
bull Oxidized pholpholipids are potent stimuli that trigger macrophage recruitment and contribute to both the initiation and progression of chronic inflammation
4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
5 Berliner JA Leitinger N and Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis J Lipid Res Suppl 50 S207ndashS212
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
THE VALUE OF LAB MEDICINE TO HEALTH CARE
bull Services provided
bull Matching blood to patient
bull Public health infectious disease identification and control
bull Rapid testing for triage and emergency care
bull Assessment of quality of care in chronic diseases
bull A basis for evidence-based medicine for cost-effective testing therapeutic treatment and interventions screening and design of therapies based in pharmacogenomics or nutrigenomics
2httpswwwfuturelabmedicineorgpdfs200720status20report20laboratory_medicine_-_a_national_status_report_from_the_lewin_grouppdfaccessed Dec 15 2012
HISTORY OF LAB SCIENCES AND INFLAMMATIONIt all started whenhellip
BRIEF HISTORY OF MEDICAL DIAGNOSIS
bull Middle ages-18th century bedside medicine
bull 1794-1848 hospital medicine
bull 1848-present laboratory medicine
bull Laboratory medicine is appreciated as the seat of modern medicine
bull Early lab techniques of diagnosis
bull Observation (palpation taste vision smell hearing)
bull Late 19th century tools Microscope thermometer stethoscope
FOUNDER OF EXPERIMENTAL PHYSIOLOGY ndash GALEN (AD
131-201)bull Combined hippocrates humoral and pythagorean
theories to correspond to various combinations of drycoldhotmoist with earthairfirewater
bull Hot + moist = blood
bull Hot + dry = yellow bile
bull Cold + moist = phlegm
bull Cold + dry = black bile
bull Ancient diagnostic methods3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
The 5 cardinal signs of inflammation originally described by Celsus 2000 years ago
Source httpwwwnaturecomnrijournalv2n10fig_tabnri915_F1html accessed March 8 2013
INFLAMMATORY MARKERS AND TESTS USED IN THE
MIDDLE AGES
bullUrineStool inspection
bull Temperature
bullPulse
bullPalpitation
bullPercussion
bullDreams3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
17TH CENTURY ndash A CENTURY OF URINE
bull William Harvey ndash blood circulation
bull 32-power microscope
led to the study of cellshistology
bull Urine tasting to differentially diagnose diabetes insipidus and diabetes mellitus
bull Urinalysis became more qualitative
bull Blood transfusions IV drug injections
bull Pulse and temperature as indicators of health3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
18TH CENTURY ndash THEORETICAL ADVANCEMENT OF MEDICINE
bull Digitalis and opium discovered for medical use
bull Septicemia was highest cause of mortality in hospitals
bull Prothrombin time and plasma prothrombin time fibrinogen (ldquocoagulable lymphrdquo) was discovered as the cause of coagulation by William Hewson (1739-1774)
bull Pulse measurement refined but rejected due to skepticism for the Galenic doctrine of there being a special pulse for every disease
bull Arterial blood pressure measurement quantified by Stephen Hales (1677-1761)
bull Discovery of sediment in the urine of febrile patientsBerger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
19TH CENTURY DATA
The microscope the X-raybull Enabled several doctors to view and discuss resultsbull Team approach to diagnosis strengthened the medical
communityrsquos claim to objective judgment
Set standards for physiology deviations and classifications
Quantitative diagnosis in the hospital laboratory allowed diagnosis of TB cholera typhoid diphtheria and syphilis
Standardized testing became the norm
Hematological testing discoveries calcium salts in the coagulation of blood mast cells white blood cell classifications
3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
INFLAMMATION GOOD BAD UGLY
bull Good necessary to promote an immune response in otherwise healthy individuals
bull Bad in allergenic and auto-immunogenic unhealthy individuals
bull Ugly chronic disease and cancer are the result of long-standing chronic inflammation
Inflammation is an incendiary process that if properly detected and treated could save lives
THE START OF INFLAMMATION
bull The start of inflammation depends on the genetics of the host and that individualrsquos response
bull Oxidized pholpholipids are potent stimuli that trigger macrophage recruitment and contribute to both the initiation and progression of chronic inflammation
4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
5 Berliner JA Leitinger N and Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis J Lipid Res Suppl 50 S207ndashS212
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
HISTORY OF LAB SCIENCES AND INFLAMMATIONIt all started whenhellip
BRIEF HISTORY OF MEDICAL DIAGNOSIS
bull Middle ages-18th century bedside medicine
bull 1794-1848 hospital medicine
bull 1848-present laboratory medicine
bull Laboratory medicine is appreciated as the seat of modern medicine
bull Early lab techniques of diagnosis
bull Observation (palpation taste vision smell hearing)
bull Late 19th century tools Microscope thermometer stethoscope
FOUNDER OF EXPERIMENTAL PHYSIOLOGY ndash GALEN (AD
131-201)bull Combined hippocrates humoral and pythagorean
theories to correspond to various combinations of drycoldhotmoist with earthairfirewater
bull Hot + moist = blood
bull Hot + dry = yellow bile
bull Cold + moist = phlegm
bull Cold + dry = black bile
bull Ancient diagnostic methods3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
The 5 cardinal signs of inflammation originally described by Celsus 2000 years ago
Source httpwwwnaturecomnrijournalv2n10fig_tabnri915_F1html accessed March 8 2013
INFLAMMATORY MARKERS AND TESTS USED IN THE
MIDDLE AGES
bullUrineStool inspection
bull Temperature
bullPulse
bullPalpitation
bullPercussion
bullDreams3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
17TH CENTURY ndash A CENTURY OF URINE
bull William Harvey ndash blood circulation
bull 32-power microscope
led to the study of cellshistology
bull Urine tasting to differentially diagnose diabetes insipidus and diabetes mellitus
bull Urinalysis became more qualitative
bull Blood transfusions IV drug injections
bull Pulse and temperature as indicators of health3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
18TH CENTURY ndash THEORETICAL ADVANCEMENT OF MEDICINE
bull Digitalis and opium discovered for medical use
bull Septicemia was highest cause of mortality in hospitals
bull Prothrombin time and plasma prothrombin time fibrinogen (ldquocoagulable lymphrdquo) was discovered as the cause of coagulation by William Hewson (1739-1774)
bull Pulse measurement refined but rejected due to skepticism for the Galenic doctrine of there being a special pulse for every disease
bull Arterial blood pressure measurement quantified by Stephen Hales (1677-1761)
bull Discovery of sediment in the urine of febrile patientsBerger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
19TH CENTURY DATA
The microscope the X-raybull Enabled several doctors to view and discuss resultsbull Team approach to diagnosis strengthened the medical
communityrsquos claim to objective judgment
Set standards for physiology deviations and classifications
Quantitative diagnosis in the hospital laboratory allowed diagnosis of TB cholera typhoid diphtheria and syphilis
Standardized testing became the norm
Hematological testing discoveries calcium salts in the coagulation of blood mast cells white blood cell classifications
3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
INFLAMMATION GOOD BAD UGLY
bull Good necessary to promote an immune response in otherwise healthy individuals
bull Bad in allergenic and auto-immunogenic unhealthy individuals
bull Ugly chronic disease and cancer are the result of long-standing chronic inflammation
Inflammation is an incendiary process that if properly detected and treated could save lives
THE START OF INFLAMMATION
bull The start of inflammation depends on the genetics of the host and that individualrsquos response
bull Oxidized pholpholipids are potent stimuli that trigger macrophage recruitment and contribute to both the initiation and progression of chronic inflammation
4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
5 Berliner JA Leitinger N and Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis J Lipid Res Suppl 50 S207ndashS212
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
BRIEF HISTORY OF MEDICAL DIAGNOSIS
bull Middle ages-18th century bedside medicine
bull 1794-1848 hospital medicine
bull 1848-present laboratory medicine
bull Laboratory medicine is appreciated as the seat of modern medicine
bull Early lab techniques of diagnosis
bull Observation (palpation taste vision smell hearing)
bull Late 19th century tools Microscope thermometer stethoscope
FOUNDER OF EXPERIMENTAL PHYSIOLOGY ndash GALEN (AD
131-201)bull Combined hippocrates humoral and pythagorean
theories to correspond to various combinations of drycoldhotmoist with earthairfirewater
bull Hot + moist = blood
bull Hot + dry = yellow bile
bull Cold + moist = phlegm
bull Cold + dry = black bile
bull Ancient diagnostic methods3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
The 5 cardinal signs of inflammation originally described by Celsus 2000 years ago
Source httpwwwnaturecomnrijournalv2n10fig_tabnri915_F1html accessed March 8 2013
INFLAMMATORY MARKERS AND TESTS USED IN THE
MIDDLE AGES
bullUrineStool inspection
bull Temperature
bullPulse
bullPalpitation
bullPercussion
bullDreams3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
17TH CENTURY ndash A CENTURY OF URINE
bull William Harvey ndash blood circulation
bull 32-power microscope
led to the study of cellshistology
bull Urine tasting to differentially diagnose diabetes insipidus and diabetes mellitus
bull Urinalysis became more qualitative
bull Blood transfusions IV drug injections
bull Pulse and temperature as indicators of health3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
18TH CENTURY ndash THEORETICAL ADVANCEMENT OF MEDICINE
bull Digitalis and opium discovered for medical use
bull Septicemia was highest cause of mortality in hospitals
bull Prothrombin time and plasma prothrombin time fibrinogen (ldquocoagulable lymphrdquo) was discovered as the cause of coagulation by William Hewson (1739-1774)
bull Pulse measurement refined but rejected due to skepticism for the Galenic doctrine of there being a special pulse for every disease
bull Arterial blood pressure measurement quantified by Stephen Hales (1677-1761)
bull Discovery of sediment in the urine of febrile patientsBerger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
19TH CENTURY DATA
The microscope the X-raybull Enabled several doctors to view and discuss resultsbull Team approach to diagnosis strengthened the medical
communityrsquos claim to objective judgment
Set standards for physiology deviations and classifications
Quantitative diagnosis in the hospital laboratory allowed diagnosis of TB cholera typhoid diphtheria and syphilis
Standardized testing became the norm
Hematological testing discoveries calcium salts in the coagulation of blood mast cells white blood cell classifications
3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
INFLAMMATION GOOD BAD UGLY
bull Good necessary to promote an immune response in otherwise healthy individuals
bull Bad in allergenic and auto-immunogenic unhealthy individuals
bull Ugly chronic disease and cancer are the result of long-standing chronic inflammation
Inflammation is an incendiary process that if properly detected and treated could save lives
THE START OF INFLAMMATION
bull The start of inflammation depends on the genetics of the host and that individualrsquos response
bull Oxidized pholpholipids are potent stimuli that trigger macrophage recruitment and contribute to both the initiation and progression of chronic inflammation
4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
5 Berliner JA Leitinger N and Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis J Lipid Res Suppl 50 S207ndashS212
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
FOUNDER OF EXPERIMENTAL PHYSIOLOGY ndash GALEN (AD
131-201)bull Combined hippocrates humoral and pythagorean
theories to correspond to various combinations of drycoldhotmoist with earthairfirewater
bull Hot + moist = blood
bull Hot + dry = yellow bile
bull Cold + moist = phlegm
bull Cold + dry = black bile
bull Ancient diagnostic methods3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
The 5 cardinal signs of inflammation originally described by Celsus 2000 years ago
Source httpwwwnaturecomnrijournalv2n10fig_tabnri915_F1html accessed March 8 2013
INFLAMMATORY MARKERS AND TESTS USED IN THE
MIDDLE AGES
bullUrineStool inspection
bull Temperature
bullPulse
bullPalpitation
bullPercussion
bullDreams3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
17TH CENTURY ndash A CENTURY OF URINE
bull William Harvey ndash blood circulation
bull 32-power microscope
led to the study of cellshistology
bull Urine tasting to differentially diagnose diabetes insipidus and diabetes mellitus
bull Urinalysis became more qualitative
bull Blood transfusions IV drug injections
bull Pulse and temperature as indicators of health3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
18TH CENTURY ndash THEORETICAL ADVANCEMENT OF MEDICINE
bull Digitalis and opium discovered for medical use
bull Septicemia was highest cause of mortality in hospitals
bull Prothrombin time and plasma prothrombin time fibrinogen (ldquocoagulable lymphrdquo) was discovered as the cause of coagulation by William Hewson (1739-1774)
bull Pulse measurement refined but rejected due to skepticism for the Galenic doctrine of there being a special pulse for every disease
bull Arterial blood pressure measurement quantified by Stephen Hales (1677-1761)
bull Discovery of sediment in the urine of febrile patientsBerger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
19TH CENTURY DATA
The microscope the X-raybull Enabled several doctors to view and discuss resultsbull Team approach to diagnosis strengthened the medical
communityrsquos claim to objective judgment
Set standards for physiology deviations and classifications
Quantitative diagnosis in the hospital laboratory allowed diagnosis of TB cholera typhoid diphtheria and syphilis
Standardized testing became the norm
Hematological testing discoveries calcium salts in the coagulation of blood mast cells white blood cell classifications
3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
INFLAMMATION GOOD BAD UGLY
bull Good necessary to promote an immune response in otherwise healthy individuals
bull Bad in allergenic and auto-immunogenic unhealthy individuals
bull Ugly chronic disease and cancer are the result of long-standing chronic inflammation
Inflammation is an incendiary process that if properly detected and treated could save lives
THE START OF INFLAMMATION
bull The start of inflammation depends on the genetics of the host and that individualrsquos response
bull Oxidized pholpholipids are potent stimuli that trigger macrophage recruitment and contribute to both the initiation and progression of chronic inflammation
4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
5 Berliner JA Leitinger N and Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis J Lipid Res Suppl 50 S207ndashS212
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
The 5 cardinal signs of inflammation originally described by Celsus 2000 years ago
Source httpwwwnaturecomnrijournalv2n10fig_tabnri915_F1html accessed March 8 2013
INFLAMMATORY MARKERS AND TESTS USED IN THE
MIDDLE AGES
bullUrineStool inspection
bull Temperature
bullPulse
bullPalpitation
bullPercussion
bullDreams3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
17TH CENTURY ndash A CENTURY OF URINE
bull William Harvey ndash blood circulation
bull 32-power microscope
led to the study of cellshistology
bull Urine tasting to differentially diagnose diabetes insipidus and diabetes mellitus
bull Urinalysis became more qualitative
bull Blood transfusions IV drug injections
bull Pulse and temperature as indicators of health3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
18TH CENTURY ndash THEORETICAL ADVANCEMENT OF MEDICINE
bull Digitalis and opium discovered for medical use
bull Septicemia was highest cause of mortality in hospitals
bull Prothrombin time and plasma prothrombin time fibrinogen (ldquocoagulable lymphrdquo) was discovered as the cause of coagulation by William Hewson (1739-1774)
bull Pulse measurement refined but rejected due to skepticism for the Galenic doctrine of there being a special pulse for every disease
bull Arterial blood pressure measurement quantified by Stephen Hales (1677-1761)
bull Discovery of sediment in the urine of febrile patientsBerger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
19TH CENTURY DATA
The microscope the X-raybull Enabled several doctors to view and discuss resultsbull Team approach to diagnosis strengthened the medical
communityrsquos claim to objective judgment
Set standards for physiology deviations and classifications
Quantitative diagnosis in the hospital laboratory allowed diagnosis of TB cholera typhoid diphtheria and syphilis
Standardized testing became the norm
Hematological testing discoveries calcium salts in the coagulation of blood mast cells white blood cell classifications
3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
INFLAMMATION GOOD BAD UGLY
bull Good necessary to promote an immune response in otherwise healthy individuals
bull Bad in allergenic and auto-immunogenic unhealthy individuals
bull Ugly chronic disease and cancer are the result of long-standing chronic inflammation
Inflammation is an incendiary process that if properly detected and treated could save lives
THE START OF INFLAMMATION
bull The start of inflammation depends on the genetics of the host and that individualrsquos response
bull Oxidized pholpholipids are potent stimuli that trigger macrophage recruitment and contribute to both the initiation and progression of chronic inflammation
4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
5 Berliner JA Leitinger N and Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis J Lipid Res Suppl 50 S207ndashS212
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
INFLAMMATORY MARKERS AND TESTS USED IN THE
MIDDLE AGES
bullUrineStool inspection
bull Temperature
bullPulse
bullPalpitation
bullPercussion
bullDreams3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
17TH CENTURY ndash A CENTURY OF URINE
bull William Harvey ndash blood circulation
bull 32-power microscope
led to the study of cellshistology
bull Urine tasting to differentially diagnose diabetes insipidus and diabetes mellitus
bull Urinalysis became more qualitative
bull Blood transfusions IV drug injections
bull Pulse and temperature as indicators of health3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
18TH CENTURY ndash THEORETICAL ADVANCEMENT OF MEDICINE
bull Digitalis and opium discovered for medical use
bull Septicemia was highest cause of mortality in hospitals
bull Prothrombin time and plasma prothrombin time fibrinogen (ldquocoagulable lymphrdquo) was discovered as the cause of coagulation by William Hewson (1739-1774)
bull Pulse measurement refined but rejected due to skepticism for the Galenic doctrine of there being a special pulse for every disease
bull Arterial blood pressure measurement quantified by Stephen Hales (1677-1761)
bull Discovery of sediment in the urine of febrile patientsBerger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
19TH CENTURY DATA
The microscope the X-raybull Enabled several doctors to view and discuss resultsbull Team approach to diagnosis strengthened the medical
communityrsquos claim to objective judgment
Set standards for physiology deviations and classifications
Quantitative diagnosis in the hospital laboratory allowed diagnosis of TB cholera typhoid diphtheria and syphilis
Standardized testing became the norm
Hematological testing discoveries calcium salts in the coagulation of blood mast cells white blood cell classifications
3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
INFLAMMATION GOOD BAD UGLY
bull Good necessary to promote an immune response in otherwise healthy individuals
bull Bad in allergenic and auto-immunogenic unhealthy individuals
bull Ugly chronic disease and cancer are the result of long-standing chronic inflammation
Inflammation is an incendiary process that if properly detected and treated could save lives
THE START OF INFLAMMATION
bull The start of inflammation depends on the genetics of the host and that individualrsquos response
bull Oxidized pholpholipids are potent stimuli that trigger macrophage recruitment and contribute to both the initiation and progression of chronic inflammation
4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
5 Berliner JA Leitinger N and Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis J Lipid Res Suppl 50 S207ndashS212
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
17TH CENTURY ndash A CENTURY OF URINE
bull William Harvey ndash blood circulation
bull 32-power microscope
led to the study of cellshistology
bull Urine tasting to differentially diagnose diabetes insipidus and diabetes mellitus
bull Urinalysis became more qualitative
bull Blood transfusions IV drug injections
bull Pulse and temperature as indicators of health3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
18TH CENTURY ndash THEORETICAL ADVANCEMENT OF MEDICINE
bull Digitalis and opium discovered for medical use
bull Septicemia was highest cause of mortality in hospitals
bull Prothrombin time and plasma prothrombin time fibrinogen (ldquocoagulable lymphrdquo) was discovered as the cause of coagulation by William Hewson (1739-1774)
bull Pulse measurement refined but rejected due to skepticism for the Galenic doctrine of there being a special pulse for every disease
bull Arterial blood pressure measurement quantified by Stephen Hales (1677-1761)
bull Discovery of sediment in the urine of febrile patientsBerger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
19TH CENTURY DATA
The microscope the X-raybull Enabled several doctors to view and discuss resultsbull Team approach to diagnosis strengthened the medical
communityrsquos claim to objective judgment
Set standards for physiology deviations and classifications
Quantitative diagnosis in the hospital laboratory allowed diagnosis of TB cholera typhoid diphtheria and syphilis
Standardized testing became the norm
Hematological testing discoveries calcium salts in the coagulation of blood mast cells white blood cell classifications
3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
INFLAMMATION GOOD BAD UGLY
bull Good necessary to promote an immune response in otherwise healthy individuals
bull Bad in allergenic and auto-immunogenic unhealthy individuals
bull Ugly chronic disease and cancer are the result of long-standing chronic inflammation
Inflammation is an incendiary process that if properly detected and treated could save lives
THE START OF INFLAMMATION
bull The start of inflammation depends on the genetics of the host and that individualrsquos response
bull Oxidized pholpholipids are potent stimuli that trigger macrophage recruitment and contribute to both the initiation and progression of chronic inflammation
4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
5 Berliner JA Leitinger N and Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis J Lipid Res Suppl 50 S207ndashS212
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
18TH CENTURY ndash THEORETICAL ADVANCEMENT OF MEDICINE
bull Digitalis and opium discovered for medical use
bull Septicemia was highest cause of mortality in hospitals
bull Prothrombin time and plasma prothrombin time fibrinogen (ldquocoagulable lymphrdquo) was discovered as the cause of coagulation by William Hewson (1739-1774)
bull Pulse measurement refined but rejected due to skepticism for the Galenic doctrine of there being a special pulse for every disease
bull Arterial blood pressure measurement quantified by Stephen Hales (1677-1761)
bull Discovery of sediment in the urine of febrile patientsBerger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
19TH CENTURY DATA
The microscope the X-raybull Enabled several doctors to view and discuss resultsbull Team approach to diagnosis strengthened the medical
communityrsquos claim to objective judgment
Set standards for physiology deviations and classifications
Quantitative diagnosis in the hospital laboratory allowed diagnosis of TB cholera typhoid diphtheria and syphilis
Standardized testing became the norm
Hematological testing discoveries calcium salts in the coagulation of blood mast cells white blood cell classifications
3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
INFLAMMATION GOOD BAD UGLY
bull Good necessary to promote an immune response in otherwise healthy individuals
bull Bad in allergenic and auto-immunogenic unhealthy individuals
bull Ugly chronic disease and cancer are the result of long-standing chronic inflammation
Inflammation is an incendiary process that if properly detected and treated could save lives
THE START OF INFLAMMATION
bull The start of inflammation depends on the genetics of the host and that individualrsquos response
bull Oxidized pholpholipids are potent stimuli that trigger macrophage recruitment and contribute to both the initiation and progression of chronic inflammation
4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
5 Berliner JA Leitinger N and Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis J Lipid Res Suppl 50 S207ndashS212
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
19TH CENTURY DATA
The microscope the X-raybull Enabled several doctors to view and discuss resultsbull Team approach to diagnosis strengthened the medical
communityrsquos claim to objective judgment
Set standards for physiology deviations and classifications
Quantitative diagnosis in the hospital laboratory allowed diagnosis of TB cholera typhoid diphtheria and syphilis
Standardized testing became the norm
Hematological testing discoveries calcium salts in the coagulation of blood mast cells white blood cell classifications
3 Berger D A brief history of medical diagnosis and the birth of the clinical laboratory Part 1 July 1999 wwwmlo-onlinecom
INFLAMMATION GOOD BAD UGLY
bull Good necessary to promote an immune response in otherwise healthy individuals
bull Bad in allergenic and auto-immunogenic unhealthy individuals
bull Ugly chronic disease and cancer are the result of long-standing chronic inflammation
Inflammation is an incendiary process that if properly detected and treated could save lives
THE START OF INFLAMMATION
bull The start of inflammation depends on the genetics of the host and that individualrsquos response
bull Oxidized pholpholipids are potent stimuli that trigger macrophage recruitment and contribute to both the initiation and progression of chronic inflammation
4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
5 Berliner JA Leitinger N and Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis J Lipid Res Suppl 50 S207ndashS212
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
INFLAMMATION GOOD BAD UGLY
bull Good necessary to promote an immune response in otherwise healthy individuals
bull Bad in allergenic and auto-immunogenic unhealthy individuals
bull Ugly chronic disease and cancer are the result of long-standing chronic inflammation
Inflammation is an incendiary process that if properly detected and treated could save lives
THE START OF INFLAMMATION
bull The start of inflammation depends on the genetics of the host and that individualrsquos response
bull Oxidized pholpholipids are potent stimuli that trigger macrophage recruitment and contribute to both the initiation and progression of chronic inflammation
4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
5 Berliner JA Leitinger N and Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis J Lipid Res Suppl 50 S207ndashS212
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
THE START OF INFLAMMATION
bull The start of inflammation depends on the genetics of the host and that individualrsquos response
bull Oxidized pholpholipids are potent stimuli that trigger macrophage recruitment and contribute to both the initiation and progression of chronic inflammation
4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
5 Berliner JA Leitinger N and Tsimikas S (2009) The role of oxidized phospholipids in atherosclerosis J Lipid Res Suppl 50 S207ndashS212
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
THE IMPORTANCE OF INFLAMMATION
bull Defined Complex non-linear set of interactions among soluble factors and cells which arise in any tissue in response to traumatic infectious post-ischemic toxic or auto-immune injury
bull Normal inflammatory process leads to recovery but if the targeted destruction and assisted repair are not controlled then inflammation can lead to persistent tissue damage by leukocytes lymphocytes or collagen
6 Carl Nathan Review article points of control in inflammation Nature 420 846-852 (19 December 2002) | doi101038nature01320
7Brown KL et al Complexities of targeting innate immunity to treat infection Trends Immunol 2007 Jun28(6)260-6 Epub 2007 Apr 30 in 1 Yoram Vodovotz Translational Systems Approaches to the Biology of Inflammation and Healing Immunopharmacol Immunotoxicol 2010 June 32(2) 181ndash195 doi 10310908923970903369867 PMCID PMC313415 NIHMSID NIHMS230084
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
WHY MEASURE INFLAMMATION
bull Inflammation and angiogenesis occur during infections auto-reactive diseases chronic inflammation as well as being hallmarks of cancer
bull Proteins are involved in the systemic response of inflammation
bull Inflammation causes cell and genome damage stimulating cellular turnover by cytokines and growth factor release and create a tissue microenvironment that can enhance invasion migration and angiogenesis 8 Karin M Lawrence T and Nizet V Innate Immunity Gone Awry Linking Microbial Infections to
Chronic Inflammation and Cancer 2006 Cell 124(4) pp823-835 Available from httpdxdoiorg101016jcell200602016 accessed Dec 8 2012
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
FIGURE 1 A GENERALIZED SCHEME TO EXPLAINTHE ORIGIN OF CHRONIC INFLAMMATION
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
THE COST OF INFLAMMATION
bull Non Communicable Diseases (NCDs) including cardiovascular diseases chronic respiratory diseases cancer and diabetes are the major global health problem of the century 9 Rosenbaum L Lamas D (2011) Facing a lsquolsquoSlow-Motion Disasterrsquorsquo - The UN Meeting on
Noncommunicable Diseases New England Journal of Medicine in PLoS One 2012 7(5) e37483 in Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
bull They are the world leading cause of disease burden and mortality are increasing in prevalence even in low-and middle-income countries the costs incurred by uncontrolled NCDs are substantial and they are an under-appreciated cause of poverty and hinder economic development 10 Bousquet J Anto J Sterk P Adcock I Chung K et al (2011) Systems
medicine and integrated care to combat chronic noncommunicable diseases Genome Medicine 3 43 in 11 Alvar Agustiacute Persistent Systemic Inflammation is Associated with Poor Clinical Outcomes in COPD A Novel Phenotype Published online 2012 May 18 doi 101371journalpone0037483 PMCID PMC3356313
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
ENVIRONMENTAL CAUSES OF INFLAMMATION
bull Environmental hypothesis for allergic and autoimmune diseasesbull Northern Geographical location high social economical
educational or occupational status stress intestinal permeability 12 Danese S Sans M Fiocchi C Inflammatory Bowel disease the role of environmental factors
Autoimmunity Reveiws 3(5) 2004 pp 394-400
bull Air Pollution plasma markers of inflammation oxidative stress and insulin resistance in children 13 Kelishadi R Mirghaffari N Poursafa P and Gidding S
Lifestyle and environmental factors associated with inflammation oxidative stress and insulin resistance in children Atherosclerosis 203 (1) 2009 311-319
bull Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction 14 Andreassi MG Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov
5
bull Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity 15 Comhair SAA Gaston BM Ricci KS Hammel J Dweik RA et al
(2011) Detrimental Effects of Environmental Tobacco Smoke in Relation to Asthma Severity PLoS ONE 6(5) e18574 doi101371journalpone0018574
bull Inflammatory Bowel disease appendectomy oral contraceptives infectionsvaccinations antibiotics childhood hygeine 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology
6(5) May 2010 pp 339-346
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
GENE X ENVIRONMENT
bull GxE interactions can be observed in the lab when cells of different genetic background respond differently when exposed to the same inflammatory environmental stimulus 4 Orozco L et al Unraveling Inflammatory Responses using Systems Genetics and
Gene-Environment Interactions in Macrophages Cell 151 658ndash670 October 26 2012
bull Play a role in the extent of inflammatory responses
bull Chronic inflammatory conditions atherosclerosis
bull The gene-environment interactions are potentially implicated in the pathogenesis of metabolic syndrome diabetes and cardiovascular disease
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
GXE
bull These genes (peroxisome proliferator activated receptor gamma alcohol dehydrogenase type 1C apo E glutathione S transferases T1 and M1)are all known to be modified by diet and lifestyle (fat diet alcohol and smoking habits)
bull There is limited understanding of the role of genetic variants of xenobiotic metabolizing enzymes and their interactions with environmental toxicants
bull There inter-individual differences in detoxification of environmental toxicants which are essential in the role of developing CVD and contribute thus to environmental cardiology as an emerging profession and the subsequent development of a personalized intervention program based upon the genomic profiling of detoxification pathways for xenobiotics 14 Andreassi MG Metabolic syndrome diabetes and atherosclerosis influence of gene-environment interaction Mutat Res 2009 Jul 10667(1-2)35-43 doi 101016jmrfmmm200810018 Epub 2008 Nov 5
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
NUTRITIONAL CAUSES OF INFLAMMATION TOO MUCH
bull Over-nutrition during gestation or lactation17 Morris MJ 2009 Early life influences on obesity risk maternal overnutrition and programming of obesity Expert Rev Endocronol Metab 4 625ndash
637 doi101586eem0945
18 Bruce KD Cagampang FR 2011 Epigenetic priming of the metabolic syndrome Toxicol Mech Methods 21 353ndash361
Both in 19 Neil A Youngson and Margaret J Morris 2013 What obesity research tells us about epigenetic mechanisms Phil Trans R Soc B 2013 368 20110337
bull Fructose and vascular inflammation independent of NF-kB fructose decreased intracellular ATP Increased NO levels and decreased both serum ICAM-1 and endothelial ICAM-1 expression (in rat kidney)
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720 doi 101681ASN2007121304 PMCID PMC2518440
bull Greater than 2x Increased risk of Crohnrsquos disease following consumption of sugarssweeteners sweets fats and oils and total fat 21 Sakamoto N Kono S Wakai K et al Dietary risk factors for inflammatory bowel disease a
multicenter case-control study in Japan Inflamm Bowel Dis 200511154ndash163 in 16 Molodecky N and Kaplan G Environmental Risk Factors for Inflammatory Bowel Disease Gastroenterology an dHepatology 6(5) May 2010 pp 339-346
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Figure A
Immunohistochemistry for ICAM-1
in 20 fructose-fed rat kidney
(a and b) Tubulointerstitium
(c and d) Glomerulus
(e and f) Arcuate artery
Figure B
Compared with rats on a control
diet (a c e) ICAM-1 expression is
significantly increased
20 Glushakova O et al Fructose Induces the Inflammatory Molecule ICAM-1 in Endothelial Cells J Am Soc Nephrol 2008 September 19(9) 1712ndash1720
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
NUTRITIONAL CAUSES OF INFLAMMATION TOO LITTLE
bull Famine in the first 10 weeks of conception causes less DNA methylation of the imprinted IGF-2 gene resulted in persistent (6 decades) epigenetic changes
bull Vitamin D status and all cause mortality22 Skaaby T Husemoen LLN Pisinger C
Joslashrgensen T Thuesen BH et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423
bull 23 Ginde AA Scragg R Schwartz RS Camargo CA (2009) Prospective study of serum 25-hydroxyvitamin D level cardiovascular disease mortality and all-cause mortality in older US adults J Am Geriatr Soc 57 1595ndash603
bull 24 Hutchinson MS Grimnes G Joakimsen RM Figenschau Y Jorde R (2010) Low serum 25-hydroxyvitamin D levels are associated with increased all-cause mortality risk in a general population the Tromso study Eur J Endocrinol 162 935ndash42
bull 25 Autier P Gandini S (2007) Vitamin D supplementation and total mortality a meta-analysis of randomized controlled trials Arch Intern Med 167 1730ndash7
bull Innate immunity via the antimicrobial peptide cathelicidin a class of antimicrobial peptides called defensinsbull Can induce expression of cathelicidin in myeloid cell
lines bronchial epithelial cells and keratinocytes 26 Adams JS Hewison M
(2008) Unexpected actions of vitamin D new perspectives on the regulation of innate and adaptive immunity Nat Clin Pract Endocrinol Metab 4 80ndash90
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Table 2 Main causes of death during follow-up in the Monica10 and Inter99 studies and vitamin D status
22 Skaaby T Husemoen LLN Pisinger C Joslashrgensen T et al (2012) Vitamin D Status and Cause-Specific Mortality A General Population Study PLoS ONE 7(12) e52423 doi101371journalpone0052423`
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
VITAMIN D STATUS AND LIVER DISEASE
bull Low levels associated with Liver disease could be a result of the disease process ndash fat malabsorption bile salt deficiency loss of absorptive surface increased intestinal permeability impaired hepatic hydroxylation reduced hepatic production of Vit D binding protein
bull Vit D supplementation in this case may not prove effective at correcting deficiency
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
LIFESTYLE CAUSES
bull InactivitySedentary Lifestyle
Various forms of physical activity decrease both acute and chronic inflammation as measured by reductions in CRP and cytokines such as TNF and IL-6
Regular physical activity is important in reducing risk for obesity and chronic diseases associated with inflammation
27 Woods JA Vieira VJ Keylock KT Exercise inflammation and innate immunity Neurol Clin 2006 Aug24(3)585-99 Review PubMed PMID 16877125
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
LIFESTYLE
bull Early life exposure to stress 29 Pervanidou P Chrousos GP 2011 Metabolic consequences of stress during childhood and adolescence Metabolism 61 611ndash619
bull Calorie Restriction and prolonged life ndash decreases inflammatory cytokines growth factors anabolic hormones and oxidative markers which are all involved in aging and cancer
30 Pallavi R Giorgio M Pelicci PG (2012) Insights into the beneficial effect of caloricdietary restriction for a healthy and prolonged life Front Physiol3 318 in
31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
ADIPOCYTOKINES AND INFLAMMATION
bull Fat tissue exerts endocrine and immune functions through the release of adipocytokines released by adipocytes leptin resistin adiponectin or visafatin as well as cytokines released by inflammatory cells infiltrating fat such as TNF- IL-6 MCP-1 (CCL-2) and IL-1
bull Effects act on immune cells to cause inflammation modulate vascular NO and SOD release mediating vascular disorders hypertension diabetes atherosclerosis and insulin resistance cancer and NASH
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Adipocytokine Immune system effects Vascular Effects
Leptin bullPro-inflammatory
bullIncrease in T cell activation
and cytokine release
proliferation
bullPromotes Th1 response
bullIncreases NK cell activation
bullIncreases macrophage
activation and cytokine
release (TNF-aIL-6 etc)
bullActivates neutrophils and
increases their chemotaxis
and oxidative burst
Prevents inflammatory
damage in conditions of
overt
immune system stimulation
bullInduces endothelial
dysfunction
bullIncreases blood pressure
bullAtherosclerosis
bullIncreases ICAM VCAM
bullPlasma levels related to
hard clinical endpoints
bullAcutely releases NO
from
endothelium
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation
And Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4)
505-528
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Adipocytokine Immune System
Effects
Vascular Effects
Resistin bullAnti-inflammatory
bullDecreases T cell
activation and
proliferation
bullInhibits NFkB
dependent cytokine
release and adhesion
molecule expression
(including TNF-aIL-6)
bullIncreases IL-10
bullInhibits phagocytosis
and oxidative burst ()
bullVasculo-protective
bullPrevents atherosclerosis
bullIs decreased in
hypertension
bullCorrelated with HDL and
bullinversely with LDL
bullPlasma levels not related
to
bullhard endpoints
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between Inflammation And
Vascular Function Review Article Journal of Physiology and Pharmacology 2006 57(4) 505-528
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Adipocytokine Immune System Effects Vascular Effects
Adiponectin bullPro-inflammatory
bullActivates NFkB dependent
cytokine release and
adhesion
molecule expression
(including TNF-aIL-6)
bullPathogenic
bullImpairs bradykinin-
dependent
vasorelaxations (NO and
EDHF)
bullNo effect on
acetylcholine
dependent
vasorelaxations
(NO)
bullVEGF and MMP up-
regulation
32 Guzik TJ Mangalat D and Korbut R Adipocytokines Novel Link Between
Inflammation And Vascular Function Review Article Journal of Physiology and
Pharmacology 2006 57(4) 505-528
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
NUTRITION TO REVERSE INFLAMMATION
bull Vitamin D for insulin sensitivity for glucose regulation
bull An enriched diet containing antioxidants such as vitamin E vitamin C β-carotene and phenolic substances has been suggested to improve symptoms by reducing disease-related oxidative stress 33 DOrazio N Gammone MA Gemello E De Girolamo M
Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull In obesity how can epigenetic modifications started or set by a high-fat diet be reversed through a reduced calorie diet or exercise
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
REVERSING THE IRREVERSIBLE
bull Hookworm protein therapy condition the immune system to prevent allergic asthma and inflammatory bowel diseases 34 Navarro S Ferreira I Loukas A The hookworm pharmacopoeia for inflammatory diseases Int J Parasitol
2012 Dec 5
bull Evidence in support of epigenetic alterations through nutritional supplementation
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
NOVEL TARGETS OF NATUROPATHIC TREATMENT AND INTERVENTION
The tissue is the issue
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35 Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Embryonic
Developme
nt
Folate DNA
Methylation
imprinting
36 Steegers-Theunissen RP et al Periconceptional maternal folic acid use of 400 microg per day is related to increased methylation of the IGF2 gene in the very young child PLoS ONE 20094e7845
37 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Choline DNA
Methylation
38 Uekawa A et al Change of epigenetic control of cystathionine beta-synthase gene expression through dietary vitamin B12 is not recovered by methionine supplementation J Nutrigenet Nutrigenomics 2009229ndash36
Protein
Restriction
DNA
Methylation
histone
modifications
39 Lillycrop KA et al Feeding pregnant rats a protein-restricted diet persistently alters the methylation of specific cytosines in the hepatic PPAR alpha promoter of the offspring Br J Nutr 2008100278ndash82
40 Lillycrop KA et al Induction of altered epigenetic regulation of the hepatic glucocorticoid receptor in the offspring of rats fed a protein-restricted diet during pregnancy suggests that reduced DNA methyl-transferase-1 expression is involved in impaired DNA methylation and changes in histone modifications Br J Nutr 2007971064ndash73
Stem Cell Butyrate Histone
acetylation
DNA
methylation
41 Mali P et al Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes Stem Cells 201028 713ndash20
Retinoic
Acid
Polycomb
Repressive
Complex 2
42 Lee ER Murdoch FE Fritsch MK High histone acetylation and decreased polycomb repressive complex 2 member levels regulate gene specific transcriptional changes during early embryonic stem cell differentiation induced by retinoic acid Stem Cells 2007252191ndash9
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 35Choi SW and Friso S
Epigenetics A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Aging Folate DNA
methylation
43 Keyes MK et al Older age and dietary folate are determinants of genomic and p16- specific DNA methylation in mouse colon J Nutr 20071371713ndash7
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunityand acute coronary syndromes Exp Gerontol 201045312ndash22
Calorie
Restriction
Histone
acetylation
45 Qiu X et al Sirtuin regulation in calorie restrictionBiochim Biophys Acta 201018041576ndash83
46 Vaquero A Sternglanz R Reinberg D NAD+-dependent deacetylation of H4 lysine 16 by class III HDACs Oncogene 2007265505ndash20
Immune
function
Folate DNA
Methylation
44 Li Y Liu Y Strickland FM Richardson B Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes Exp Gerontol 201045312ndash22
Inflammation Resveratrol Histone
acetylation
47 Sanchez-Fidalgo S et al Dietary supplementation of resveratrol attenuates chronic colonic inflammation in mice Eur J Pharmacol 201063378ndash84
48 Donnelly LE et al Anti-inflammatory effects of resveratrol inlung epithelial cells molecular mechanisms Am J Physiol Lung Cell Mol Physiol 2004287L774ndash83
AdoMet Histone
acetylation
49 Ara AI et al S-adenosylmethionine inhibits lipopolysaccharide-induced gene expression via modulation of histone methylation Hepatology 2008471655ndash66
Methyl-
deficient
diet
mRNA50 Pogribny IP et al Difference in expression of hepatic microRNAs miR-29c miR-34a miR-155 and miR-200b is associated with strainspecific susceptibility to dietary nonalcoholic steatohepatitis in mice Lab Invest Epub 2010 Jun 14
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics
A new bridge between nutrition and health Advances in Nutrition 2010 11-16
Cancer Methyl-
deficient
diet
Histone
modification
mRNA
51 Pogribny IP et al Histone H3 lysine 9 and H4 lysine 20 trimethylation and the expression of Suv4ndash20h2 and Suv-39h1 histone methyltransferases in hepatocarcinogenesis induced by methyl deficiency in rats Carcinogenesis 2006271180ndash6
52 Tryndyak VP et al Down-regulation of the microRNAs miR-34a miR-127 and miR-200b in rat liver during Hepato-carcinogenesis induced by a methyl-deficient diet Mol Carcinog 200948479ndash87
Genistein DNA
Methylation
mRNA
53 Qin W et al Soy isoflavones have an antiestrogenic effect and alter mammary promoter hypermethylation in healthy premenopausal women Nutr Cancer 200961238ndash44
54 Tang WY et al Persistent hypomethylation in the promoter of nucleosomal binding protein 1 (Nsbp1) correlates with over-expression of Nsbp1 in mouse uteri neonatally exposed to diethylstilbestrol or genistein Endocrinology 20081495922ndash31
55 Salerno E et al Correcting miR-15a16 genetic defect in New Zealand Black mouse model of CLL enhances drug sensitivity Mol Cancer Ther 200982684ndash92
Inhibiting
EGC-3G
DNA
methylation
PRC
56 Fang M Chen D Yang CS Dietary polyphenols may affect DNA methylation J Nutr 2007137S223ndash8
57 Balasubramanian S Adhikary G Eckert RL The Bmi-1 polycomb protein antagonizes the (2)-epigallocatechin-3-gallate-dependent suppression of skin cancer cell survival Carcinogenesis 201031496ndash503
Curcumin mRNA58 Sun M Estrov Z et al Curcumin (diferuloylmethane) alters the expression profiles of microRNAs in human pancreatic cancer cells Mol Cancer Ther 20087464ndash73
59 Yang J Cao Y Sun J Zhang Y Curcumin reduces the expression of Bcl- 2 by upregulating miR-15a and miR-16 in MCF-7 cells Med OncolEpub 2009 Nov 12
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Process or
Condition
Nutrient or
Diet
Epigenetic
Mechanism
References in 37 Choi SW and Friso S Epigenetics A
new bridge between nutrition and health Advances in Nutrition 2010 11-16
Obesity Insulin
Resistance
High-fat
diet
DNA
methylation
mRNA
60 Widiker S Karst S Wagener A Brockmann GA High-fat diet leads to a decreased methylation of the Mc4r gene in the obese BFMI and the lean B6 mouse lines J Appl Genet 201051193ndash7
61 Cirera S Birck M Busk PK Fredholm M Expression profiles ofmiRNA-122 and its target CAT1 in minipigs (Sus scrofa) fed a high cholesterol diet Comp Med 201060136ndash41
Methyl-
deficient
diet
DNA
methylation
62 Sinclair KD et al DNA methylation insulin resistance and blood pressure in offspring determined by maternal periconceptional B vitamin and methionine status Proc Natl Acad Sci USA 200710419351ndash6
Curcumin Histone
acetylation
63 Chiu J Khan ZA Farhangkhoee H Chakrabarti S Curcumin prevents diabetes-associated abnormalities in the kidneys by inhibiting p300 and nuclear factor-kappaB Nutrition 200925964ndash72
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
NATUROPATHIC APPROACHES TO QUELLING
INFLAMMATIONbull Ginger
bull Diabetes Hypertension 64 Al-Amin ZM Thomson M Al-Qattan KK Peltonen-Shalaby
R Ali M Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats Br J Nutr 2006 Oct96(4)660-6 PubMed PMID 17010224
bull Osteoarthritis 65 Shen CL Hong KJ Kim SW Effects of ginger (Zingiber officinale Rosc) on decreasing the
production of inflammatory mediators in sow osteoarthrotic cartilage explants J Med Food 2003 Winter6(4)323-8 PubMed PMID 14977440
bull Anti-thrombotic and anti-inflammatory 66 Thomson M Al-Qattan KK Al-Sawan SM Alnaqeeb MA Khan I Ali M The use of ginger (Zingiber officinale Rosc) as a potential anti-inflammatory and antithrombotic agent Prostaglandins Leukot Essent Fatty Acids 2002 Dec67(6)475-8 PubMed PMID 12468270
67 Nicoll R Henein MY Ginger (Zingiber officinale Roscoe) a hot remedy for cardiovascular disease Int J Cardiol 2009 Jan 24131(3)408-9 Epub 2007 Nov 26 Review PubMed PMID 18037515
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
QUELLING INFLAMMATION
bull Marine omega-3 polyunsaturated fatty acids bioactive peptides isolated from fish protein hydrolysates as well as algal fucans galactans and alginates have been shown to possess anticoagulant anticancer and hypocholesterolemic activities Crustaceans and seaweeds contain powerful antioxidants such as carotenoids and phenolic compounds
69 Lordan S Ross RP Stanton C Marine bioactives as functional food ingredients potential to reduce the incidence of chronic diseases Mar Drugs 20119(6)1056-100 doi 103390md9061056
bull Indications
bull Diabetes heart disease IBS Alzheimerrsquos disease Parkinsonrsquos disease allergies asthma cancer 33 DOrazio N Gammone
MA Gemello E De Girolamo M Cusenza S Riccioni G Marine bioactives pharmacological properties and potential applications against inflammatory diseases Mar Drugs 2012 Apr10(4)812-33
bull Rheumatoid Arthritis 70 Miles EA Calder PC Influence of marine n-3 polyunsaturated fatty acids on immune
function and a systematic review of their effects on clinical outcomes in rheumatoid arthritis Br J Nutr 2012 Jun107 Suppl 2S171-84
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
QUELLING INFLAMMATION
bull Curcumin and human cell line studies
bull Anti-cancer
bull Inh cyclin D1 (colon cancer)
bull Inh MMP aminopeptidase N (melanoma fibrosarcoma breast cancer intestinal epithelial)
bull Inh HIF-1 (hepatocellular carcinoma)71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
QUELLING INFLAMMATION
bull Curcumin
bull Anti-inflammatory
bull Inh NF-kB (myeloid leukemia B non-Hodgkinrsquos Lymphoma embryonic kidney)
bull Inh COX-2 (intestinal microvascular endothelial colonic epithelial microglial)
bull IL-1 IL-6 IL-8 TNF- (Esophageal epithelial head and neck cancer)
bull Down-regulates MMP2 and MMP9 invasion migration and production in human fibrosarcoma cells
72Yodkeeree S Garbisa S amp Limtrakul P (2008) Tetrahydrocurcumin inhibits HT1080 cell migration and invasion via downregulation of MMPs and uPA Acta Pharmacol Sin 29 853ndash860 in
71 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
CLINICAL EFFECTS OF CURCUMIN HUMAN TRIALS
CURCUMIN
67
Type 2 Diabetes Atherosclerosis Raised serum Cholesterol
Alzheimerrsquos Disease
Familial Adenomatous
Polyposis Cancers
Irritable Bowel Syndrome
Functional Biliary Disorders
Tropical Pancreatitis
Gastric Ulcers
Rheumatoid Arthritis
Psoriasis
Inflammatory eye disease
Renal Graft function Post-
Operative inflammation
Inflammatory Bowel
Disease
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Inflammation Pathway and
Resulting Condition
Naturopathic
Prevention or
Treatment Options
References in 72 Epstein J Sanderson I and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Stress response ndashgt heat
shock protein (HSP) to
protect cells from stress
Alzheimerrsquos
Vitamin E and Phenolic
substances which
promote anti-oxidation
and chemo-
prevention induce
HSP response1
73 Calabrese V Butterfield DA Stella AM Nutritional antioxidants and the heme oxygenase pathway of stress tolerance novel targets for neuroprotection in Alzheimers disease Ital J Biochem 2003 Dec52(4)177-81 Review PubMed PMID 15141484
Prostaglandin production
via COX-1 or COX-2 -
Arthritis
Modulate genetic
expression of NF-kB1
Ginger2
Thunder God Vine
Tryptergium wilfordii3
74 Terracciano S Aquino M Rodriquez M Monti MC Casapullo A Riccio R Gomez-Paloma L Chemistry and biology of anti-inflammatory marine natural products molecules interfering with cyclooxygenase NF-kappaB and other unidentified targets Curr Med Chem 200613(16)1947-69 Review
68 Grzanna R Lindmark L Frondoza CG Ginger--an herbal medicinal product with broad anti-inflammatory actions J Med Food 2005 Summer8(2)125-32 Review PubMed PMID 16117603
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 75 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Inflammation Pathway
and Resulting Condition
Naturopathic Prevention
or Treatment Options
References in 72 Epstein J Sanderson I
and MacDonald T (2010) Curcumin as a therapeutic agent the evidence from in vitro animal and human studies British Journal Of Nutrition 103 (11) 1545-1557
Increased Plasma IL-6
and TNF-alpha
Aging
Increased morbidity in
elderly people
Type 2 diabetes
Atherosclerosis
Alzheimerrsquos Disease
Regular physical
activity26 74
Regular sleep75
Weight loss via exercise
related metabolic
changes - induce IL-6
and induce lipolysis and
hepatic glucose output76
28 Bruunsgaard H Physical activity and modulation of systemic low-level inflammation J Leukoc Biol 2005 Oct78(4)819-35 Epub 2005 Jul 20 Review PubMed PMID 16033812
74 Bruunsgaard H Andersen-Ranberg K Jeune B Pedersen AN Skinhoslashj P Pedersen BK A high plasma
concentration of TNF-alpha is associated with dementia in centenarians J Gerontol A Biol Sci Med Sci 1999 Jul54(7)M357-64 PubMed PMID 10462168
75 Okun ML Reynolds CF 3rd Buysse DJ Monk TH Mazumdar S Begley A Hall M Sleep variability health-related practices and inflammatory markers in a community dwelling sample of older adults Psychosom Med 2011 Feb-Mar73(2)142-50
76 Pedersen BK Steensberg A Schjerling P Exercise and interleukin-6 Curr OpinHematol 2001 May8(3)137-41 Review PubMed PMID11303145
NF-kappaB -gt TNF-Alpha
IL-6
Aging
Chronic inflammation
Atherosclerosis
Type 2 Diabetes
Zinc turnover via
Metallothionine
homeostasis ndash meat and
high zinc containing
foods1
77Vasto S Mocchegiani E Malavolta M Cuppari I Listigrave F Nuzzo D Ditta V Candore G Caruso C Zinc and inflammatoryimmune response in aging Ann N Y Acad Sci 2007 Apr1100111-22 Review
C-reactive Protein
Obesity
Insulin resistance
Metabolic syndrome
Weight loss increased
physical activity
Metformin1
Alteration in dietary
composition2
carbohydrate restricted
78 Levy J Cobas RA and Gomes MB Asessment of efficacy and tolerability of once-daily extended release metformin in patients with type 2 diabetes mellitus Diabetes 5451566 -1572
79 Beavers KM Nicklas BJ Effects of lifestyle interventions on inflammatory markers in the metabolic syndrome Front Biosci (Schol Ed) 2011 Jan 13168-77 Review
80 Al-Sarraj T Saadi H Calle MC Volek JS Fernandez ML Carbohydrate restriction as a first-line dietary intervention effectively reduces biomarkers of metabolic syndrome in Emirati adults J Nutr 2009 Sep139(9)1667-76
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
INFLAMMATORY DEBATES
Genomics vs Epigenetics vs Proteomics
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
WHY BIOMARKERS
bull Biomarkers have the potential to enable diagnosis before the onset of symptoms predict specific organ involvement and disease flares identify clinically meaningful disease subsets predict and monitor response to therapy and describe organ or tissue damage
bull Can be useful in identifying genetic predisposition to disease or environmental triggers
bull May provide early information about potential efficacy of experimental agents or mechanisms underlying drug activity
bull Can become validated surrogate endpoints for prevention trials
86 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 54 httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
CURRENT MARKERS EPIGENETIC VS PROTEOMIC
VALUEbull Proteomics show functional disease state since proteins
(and not transcripts) represent current status 81 Stroncek DF Burns C
Martin BM Rossi L Marincola FM Panelli MC Review Advancing cancer biotherapy with proteomics J Immunother 2005 May-Jun 28(3)183-92
bull Epigenetics have causal status
The two can be combined for patient care depending on the goalpriority
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
DEFINITIONS PROTEOMICS
bull Molecular patterns via protein analysis functional analysis
bull Uses immunoassays ELISA MSIA SISCAPA to assess complex biologic processes such as immunotherapy for cancer
bull Acute phase reactant proteins
bull At present a comprehensive proteomics signature is not achievable due to the highly variable protein concentrations the biochemical diversity of proteins and the need for better profiles to measure the significant fraction of the proteome 82 Kikuchi T Carbone DP Proteomics analysis
in lung cancer challenges and opportunities Respirology 2007 Jan12(1)22-8 Review PubMed PMID 17207021
bull Difficulty with reproducibility across different labs
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
DEFINITIONS EPIGENETICS
bull Transcripts of DNA or mRNA levels do not reflect expression activity due to post-translational modifications ubiquitination protease cleavage glycosylation phosphorylation methylation and acetylation
Bottom line Protein concentration (and activity) may be different than gene expression due to post-translational modification
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
DEFINITIONS GENOMICS
bull Functional Genomics
bull Computational Genomics
bull Personal Genomics
bull Epigenomics
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
BIOMARKERS PROTEOMICS
bull Acute phase reactant proteins (APRP)
bull CRP and hsCRP
bull Ferritin hepcidin
bull Leukocyte Global DNA Methylation
bull F2-Isoprostanes Hp-beta Ccl-18
bull Fasting insulin
bull IL-6
bull MMP-2 and MMP-9
bull α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3 and C4B and retinol binding protein
bull LRG1
bull IA-2 and IA-2 beta gene anti-bodies endogenous C-peptide
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Biomarker Name Condition Notes
Acute phase reactant
proteins (APRP)
Cancer APRP expressions
distinct for type
subtype and stages
CRP and hsCRP bacterial infection rheumatic
fever active arthritis myocardial
infarction malignancies and in
the post-operative state
Ferritin hepcidin inflammation liver disease
chronic infection autoimmune
disease and some types of
cancer
Leukocyte Global DNA
Methylation
cancer cardiovascular disease
pediatric syndromes autoimmune
disease and genetic disorders
F2-Isoprostanes atherosclerosis (obesity and
cigarette smoking)
hypercholesterolemia diabetes
HIV asthma neurodegenerative
RA and certain cancer types
83 Pang WW Abdul-Rahman PS Wan-Ibrahim WI and Hashim OH Can the acute phase reactant proteins be used as cancer
biomarkers Int J Biol Markers 2010 25(1) 1-11
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
CRP AND HS CRP
bull Increased CRP levels are found in inflammatory conditions including bacterial infection rheumatic fever active arthritis myocardial infarction malignancies and in the post-operative state 84
httpwwwquestdiagnosticscomtestcenterTestDetailactionntc=4420 accessed Jan 6 2013
bull A high-sensitivity CRP (hs-CRP) test may be used by itself in combination with other cardiac risk markers or in combination with a lp-PLA2 test that evaluates vascular inflammation
bull 85 httplabtestsonlineorgunderstandinganalyteshscrptabtest accessed Jan 6 2013
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
FERRITIN AND HEPCIDIN
bull Ferritin is an acute phase reactant and can be increased in inflammation liver disease chronic infection autoimmune disease and some types of cancer Damage to the liver spleen and bone marrow can cause elevation of ferritin with normal iron results 86
httplabtestsonlineorgunderstandinganalytesferritintabtest accessed Jan 6 2013
bull Hepcidin is an iron-regulatory hormone Its receptor and iron channel ferroportin control absorption storage and distribution of iron
bull Hepcidin function is protectively altered during infections and inflammation to decrease iron availability to invading pathogens and may cause or contribute to the anemia associated with infectious and inflammatory disorders 87 Ganz T Hepcidin and iron regulation 10 years later Blood 2011 1174425-4433
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
EPIGENETICS LEUKOCYTE GLOBAL DNA METHYLATION
PROFILEbull DNA methylation of cytosine followed by guanine
(CpG nucleotides) is the most widely studied epigenetic markerbull Implications in the control of embryo development cell
differentiation X chromosome inactivation in females genomic imprinting suppression of endogenous retroviruses and chromosomal instability
bull Abnormalities in DNA methylation patterns associated with disease etiologies cancer cardiovascular disease pediatric syndromes autoimmune disease and genetic disorders
bull In the aging process via specific hypermethylation of the promoter regions of various genes
88 Portela A Esteller M (2010) Epigenetic modifications and human disease Nat Biotechnol 28(10) 1057ndash1068
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
EPIGENETICS DNA METHYLATION
Recent studies indicate the potential for using global DNA methylation profiles of peripheral blood leukocytes as a suitable biomarker for cancer 89 Woo HD Kim J (2012) Global DNA hypomethylation in
peripheral blood leukocytes as a biomarker for cancer risk a meta-analysis PLoS One 7(4) e34615
Bladder 90 Cash HL Tao L Yuan JM Marsit CJ Houseman EA et al (2012) LINE-1 hypomethylation is associated with bladder cancer risk
among nonsmoking Chinese Int J Cancer 130(5) 1151ndash1159
Breast 91 Choi JY James SR Link PA McCann SE Hong CC et al (2009) Association between global DNA hypomethylation in leukocytes and
risk of breast cancer Carcinogenesis 30(11) 1889ndash1897
Renal cell 92 Liao LM Brennan P van Bemmel DM Zaridze D Matveev V et al (2011) LINE-1 methylation levels in leukocyte DNA and risk of
renal cell cancer PLoS One 6(11) e27361
In 31 Gomes MVM Toffoli LV Arruda DW Soldera LM Pelosi GG et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
CORRELATION WITH LAB PARAMETERS
bull DNA methylation profile of leukocytes showed a statistical correlation to the mean level of total protein (p = 00004) alpha 2 globulin (p = 00013) and albumin (p = 00015)
bull The mean global leukocyte DNA methylation of the elderly population showed a statistical association with daily ingestion of carbohydrates (p = 00494) lipids (p = 00494) vitamin B6 (p = 00421) and magnesium (p = 00302)
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Table 1 Correlation between global DNA methylation and clinical laboratorial values
31 Gomes MVM Toffoli LV Arruda DW Soldera LM et al (2012) Age-Related Changes in the Global DNA Methylation Profile of Leukocytes Are Linked to Nutrition but Are Not Associated with the MTHFR C677T Genotype or to Functional Capacities PLoS ONE 7(12) e52570 doi101371journalpone0052570 httpwwwplosoneorgarticleinfodoi101371journalpone0052570
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
BIOLOGICAL PROCESS MARKERS FOR INFLAMMATION AND CANCER
bull Inflammation F2-Isoprostane Hpβ Ccl8 and Fasn
bull Angiogenesis thrombospondin 1 and 4 renin 1 and 2 platelet factor 4 trefoil factor 1 and collagen binding protein 2
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
INFLAMMATION F2-ISOPROSTANES
bull Isoprostanes are the most important biomarker of oxidative stress in human disease
94 Lawson JA Rokach J Fitzgerald GA (1999) Isoprostanes formation analysis and use as indices of lipid peroxidation in vivo J Biol Chem 27424441ndash24444
95 Montuschi P Barnes P Roberts JL (2004) Isoprostanes markers and mediators of oxidative stress The FOSEB J 181792ndash1796
96 Milne GL Musiek ES Morrow JD (2005) F2-Isoprostanes a marker of oxidative stress in vivo Biomarkers 1S10ndashS23
97 Milne GL Yin H Brooks JD Sanchez S Jackson RL Morrow JD (2007) Quantification of F2-isoprostanes in biological fluids and tissues as a measure of oxidant stress Methods Enzymol 433113ndash126
98 Brys M et al 2013 Relationship of urinary isoprostanes to prostate cancer occurrence Mol Cell Biochem 372149-153
bull Isoprostanes are a prostaglandin-like molecule They are formed from peroxidation of arachidonic acid DHA EPA and adrenic acid They should not be measured by radio-immunoassay but by GCMS
bull Urinary F2-Isoprostanes validated biomarker of oxidative stress increased risk of prostate cancer independent of central adiposity or obesity PSA family history and smoking history
99 Barocas D et al 2011 Oxidative stress measured by Urine F2-Isoprostane level is associated with prostate cancer J Urol 185(6) 2102-2107
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
METABOLISM OF F2-ISOPROSTANES
bull Initially formed from arachidonic acid esterified in tissue phospholipids and then released as free fatty acids by phospholipase
bull Circulate in plasma metabolized in the liver filtered in the kidney and appear in urine
bull Diseases associated with increased F2-Isoprostanes
bull atherosclerosis (obesity and cigarette smoking)
bull hypercholesterolemia
bull diabetes
bull HIV
bull asthma
bull neurodegenerative
bull RA
bull and certain cancer types100 Milne GL Et al 2011 Isoprostane Generation and Function Chem Rev 111(10) 5073-5996
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Biomarker Name Condition Notes
Haptoglobin beta Hgβ Lung cancer
Infiltrating ductal carcinoma
Epithelial ovarian cancer
Sepsis ndash need
addrsquol CRP
CCL-18 Bladder Ovarian and Cervical
cancer
Fasting Insulin Insulin Resistance
IL-6 Post op infection
MM survival time
Cardiovascular w CRP
Osteosarcoma
Gastric Cancer
Multiple Myeloma
RA and Diabetes
Insulin Resistance
normal value
lt0-5pgml
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
HAPTOGLOBIN BETA CHAIN (HP)
Supportive biomarker for prognostic value and overall survival
bull human lung cancer101 Kang SM et al 2011 The Haptoglobin beta chain as a supportive biomarker for human lung cancers Mol Biosyst 7(4) 1167-75
bull prostate cancer 102 Fujimura T et al Glycosylation status of haptoglobin in sera of patients with prostate cancer vs benign prostate disease or normal
subjects Int J Cancer 2008 Jan 1122(1)39-49
bull epithelial ovarian cancer 103 Zhao C et al 2007 Circulating Haptoglobin is an Independent Prognostic Factor in the Sera of Patients with Epithelial Ovarian Cancer Neoplasia January 9(1) 1-7
During times of acute inflammation and requires other tests (CRP) to confirm sepsis infection etc because generally it is a test for hemolytic anemia and extravascular hemolysis
Can be increased with use of the following drugs androgens corticosteroids
Can be decreased with the following drugs isoniazid chlorpromazine quinidine streptomycin nitrofurantoin and birth control pills
104 httplabtestsonlineorgunderstandinganalyteshaptoglobintabtest accessed Jan 20 2013
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
INFLAMMATION CCL-18
bull CCL 18 C-C motif chemokine 18
bull Bladder cancer when used with nuclear matrix protein-22 and bladder tumor antigen Cystoscopy still gold standard
105 Urquidi V Kim J Chang M Dai Y Rosser CJ et al (2012) CCL18 in a Multiplex Urine-Based Assay for the Detection of Bladder Cancer PLoS ONE 7(5) e37797 doi101371journalpone0037797
bull Ovarian cancer 106 Zohny SF Fayed ST (2010) Clinical utility of circulating matrix metalloproteinase-7 (MMP-7) CC chemokine ligand 18 (CCL18) and CC chemokine ligand 11 (CCL11) as markers for diagnosis of epithelial ovarian cancer Med Oncol 27 1246ndash1253 doi 101007s12032-009-9366-x
107 Wang Q Li D Zhang W Tang B Li QQ et al (2011) Evaluation of proteomics-identified CCL18 and CXCL1 as circulating tumor markers for differential diagnosis between ovarian carcinomas and benign pelvic masses Int J Biol Markers
bull Cervical cancer108 Rajkumar T Sabitha K Vijayalakshmi N Shirley S Bose MV et al (2011) Identification and validation of genes involved in cervical tumourigenesis BMC Cancer 11 80 doi 1011861471-2407-11-80
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
BIOMARKER FASTING INSULIN
bull Obesity related Insulin resistance and diabetes ndash dietary leucine as an environmental modifier acting on multiple metabolites stimulated phosphorylation of p70S6 Kinase (indicates inc levels of mTOR normally associated with insulin resistance)
bull 8 week old male mouse model design mouse chow and high fat diets with or without supplemental leucine (15wv) in water x 8 weeks
bull Prevents hepatic steatosis reduces obesity induced adipose tissue inflammation rescues insulin signaling in muscle liver and fat
bull Mouse dose 130mgdaymouse
109 Macotela Y Emanuelli B Baringng AM Espinoza DO Boucher J et al (2011) Dietary Leucine - An Environmental Modifier of Insulin Resistance Acting on Multiple Levels of Metabolism PLoS ONE 6(6) e21187 doi101371journalpone0021187
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
BIOMARKER IL-6
bull Pleotropic Cytokine normal value lt0-5pgml 110 httpwwwaruplabcomguidesugtests0051537jsp accessed Jan 26 2013
bull Diagnostically most accurate for detecting intraoperative prosthetic joint infection in comparison to WBC ESR CRP and the time of revision arthroplasty
111 Berbari E Mabry T Tsaras G Spangehl M Erwin PJ Murad MH Steckelberg J Osmon D Inflammatory blood laboratory levels as markers of prosthetic joint infection a systematic review and meta-analysis J Bone Joint Surg Am 2010 Sep 192(11)2102-9
bull Prognostic significance for survival time in multiple myeloma patients if IL-6 was above 7pgml
bull 50 survival rate if below 7pgml was 537 months compared to 27 months if above 7pgml
112 Ludwig H Nachbaur DM Fritz E Krainer M and Huber H Interleukin-6 is a prognostic factor in multiple myeloma (letter) 1991 Blood 77 2794-2795
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
CRP AND IL-6 TRANS FATS
bull CRP and IL-6 are independent predictors of
cardiovascular disease in healthy women Nursesrsquo Health Study I cohort 730 healthy women followed between 1989-1990 CRP was 73 higher in those with the highest trans fat intake IL-6 was 17 higher
bull Trans fats adversely affect endothelial function as measured by CRP sTNFR E-selectin sICAM-1 and sVCAM-1 regardless of fiber cholesterol Vitamin E intake and glycemic load
113 Blake G J amp Ridker P M (2003) C-reactive protein and other inflammatory risk markers in acute coronary syndromes J Am Coll Cardiol 41 37Sndash42S In
114 Lopez-Garcia E et al Nutiritonal Epidemiology Consumption of Trans Fatty Acids is Related to Plasma Biomarkers of Inflammation and Endothelial Dysfunction Journal of Nutrition March 2005 135(3) 562-566
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
CORONARY HEART DISEASE MARKERS
NURSES HEALTH STUDY AND HEALTH PROFESSIONALS FOLLOW UP STUDY
bull Plasma soluble TNF receptors 1 and 2
bull CRP
bull IL-6
Better predictors when combined not individually assessed
Increased risk of CHD with combined elevations in CRP and IL-6 for both sexes but elevated TNF levels were significant among women
115 Pai JK et al Inflammatory markers and the risk of coronary heart disease in men and womenN Engl J Med 2004 Dec 16351(25)2599-610
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
INHIBITION OF IL-6 CLINICAL VALUE
bull In RA
bull IL-6 is involved in the pathogenesis of RA development via TNF
118 Hennigan S and Kavanaugh A Interleukin-6 inhibitors in the treatment of rheumatoid arthritis Ther Clini Risk Manag 2008 4(4) 767-775
bull Inhibiting IL-6 signaling positively affected insulin sensitivity and lipoprotein A levels in diabetic patients with RA Long term elevated IL-6 can be causal in the pathogenesis of insulin resistance
119 Schultz O Oberhauser F Saech J Rubbert-Roth A Hahn M et al (2010) Effects of Inhibition of Interleukin-6 Signaling on Insulin Sensitivity and Lipoprotein (A) Levels in Human Subjects with Rheumatoid Diseases PLoS ONE 5(12) e14328 doi101371journalpone0014328
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Biomarker Name Condition Notes
MMP-2 and MMP-9 Arthritis type 1 collagen
degradation in cartilage
α1-microglobulin fibrinogen
Apo lipoproteins A and E
complement C3 and C4B and
retinol binding protein
Systemic Auto-Immune
Disease
Leucine Rich α2 Glycoprotein
LRG1α
Multiple Sclerosis Twin studies
serum and
CSF fluid
elevations
Auto-antibodies to human IA-2
and IA-2 beta C- Peptide
Type 1 Diabetes
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
BIOMARKERS MMP-2 FOR ARTHRITIS
bull MMP-2 (gelatinase A) and MMP-9 (gelatinase B) in type 1 collagen degradation in cartilage
bull In RA patients synovial fluids include increased levels of MMP-2 and MMP-9 but MMP-2 has a dual role and MMP-9 has a direct inflammatory role
93 Kelly-Spratt KS Pitteri SJ Gurley KE Liggitt D Chin A et al (2011) Plasma Proteome Profiles Associated with Inflammation Angiogenesis and Cancer PLoS ONE 6(5) e19721 doi101371journalpone0019721
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
BIOMARKERS SYSTEMIC AUTOIMMUNE DISEASE
bull SLE
bull Juvenile idiopathic arthritis
bull Juvenile dermatomysitis
All share common alterations of plasma protein expression and molecular pathways α1-microglobulin fibrinogen Apo lipoproteins A and E complement C3a and C4b and retinol binding protein
120 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
Chronic Lyme disease and AIDS C4a levels highest
121 Stricker RB Savely VR Motanya NC and Giclas PC Complement split products C3a and C4a in chronic lyme disease (2009) Scand J Immunol 69(1) 64-69
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
MULTIPLE SCLEROSIS
bull Leucine-rich α2 glycoprotein marker (LRG1) - a molecule involved in signal transduction cell adhesion and granulocyte differentiation - was elevated in plasma from affected twins and was also found elevated in both the cerebrospinal fluid and serum proteomes from multiple sclerosis patients
121 OConnor KC et al Comprehensive phenotyping in multiple sclerosis discovery based proteomics and the current understanding of putative biomarkers Dis Markers 2006 22213-225 PMID 17124343 in 121 Terrance P OHanlon1 et al Plasma proteomic profiles from disease-discordant monozygotic twins suggest that molecular pathways are shared in multiple systemic autoimmune diseases Arthritis Research amp Therapy 2011 13R181 doi101186ar3506
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
TYPE 1 DIABETES
bull Auto-antibodies to the human genes IA-2 and IA-2 beta appear years before the development of clinically apparent type 1 diabetes and therefore can serve as predictive markers for this disease
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 62
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
bull Endogenously produced C-peptide secretion is the hallmark for monitoring beta cell function and preserving beta cell function equates to normal glycemic control
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 69
httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
SUMMARY
bull Brief history of lab medicine
bull Causes of inflammation
bull GxE
bull Nutritional
bull Lifestyle
bull Body Compositional
Stopping inflammation with novel targeted therapies used in naturopathic medicine
Genomics vs Epigenetics vs Proteomics as biomarkers in evaluating diagnostic and treatment status of various inflammatory conditions
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Process or
Condition
Naturopathic
Approaches
Proteomic Diagnostic
Marker
References
Lung cancer Vitamin D3
Melatonin EPA
Haptoglobin chain
serum amyloid A and
Apo lipoprotein A-1
122 Indovina P Marcelli E Maranta P and Tarro G 2011 Lung Cancer Proteomics Recent Advances in Biomarker Discovery Int J Proteomics 2011 doi 1011552011726869
Infiltrating ductal
breast
carcinoma
Melatonin EPA
EGCG Vitamin
D3
1-antitrypsin
haptoglobin
precursors
123 Hamrita B Chahed K Trimeche M et al Proteomics-based identification of α1-antitrypsin and haptoglobin precursors as novel serum markers in infiltrating ductal breast carcinomas Clinica Chimica Acta 2009404(2)111ndash118
Epithelial
Ovarian Cancer
Haptoglobin103 Zhao C Annamalai L Guo C Kothandaraman N Koh SC Zhang H Biswas A Choolani M Circulating haptoglobin is an independent prognostic factor in the sera of patients with epithelial ovarian cancer Neoplasia 2007 Jan9(1)1-7
Type 1 Diabetes Omega-3 fatty
acids
Vitamin D3
20 Progress in Autoimmune Diseases Research NIH Autoimmune Disease Coordinating Committee Report to Congress March 2005 p 73httpwwwniaidnihgovtopicsautoimmuneDocumentsadccfinalpdf accessed Dec 16 2012
Rheumatoid
Arthritis
Insulin
Resistance
Curcumin Green
Tea and
Resveratrol
IL-6124 Gonzales AM and Orlando RA Curcumin and Resveratrol inhibit NFK-B mediated cytokine expression in adipocytes 2008 Nutrition and Metabolism 517
Type 2 Diabetes Leucine Vitamin
D3
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom
Process or
Condition
Naturopathic
Approaches
Proteomic
Diagnostic Marker
References in
125 Zamperion ER Kamhi EJ 2012 Natural Support for Autoimmune and Inflammatory Disease J Rest Med 138-47
Cox-2 inhibition
RA
Hops iso-alpha
acids with
oleanolic acid and
rosmarinic acid
126 Lukazer D Darland G Tripp M et al A pilot trial evaluation Meta050 a proprietary combination of reduced iso-alpha acids rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia Phytotherapy research 2005 19(10)864-869
SLE RA
borelliosis and
co-infections
Artemesia127 Zhang YX Sun HX Immunosuppresive effect of ethanol extract of Artemesia annua on specific antibody and cellular responses of mice against ovalbumin Immunopharmacol Immunotoxicol 200931(4)625-30
Autoimmune
disease
Cancer
RA
Reishi Mushroom IL-18 TNFα 128 Vanderhem LG Vandervliet JA Bocken CFM et al Studies of a new immunemodulating agent Transplantation 1995 60(5) 438-443129 Bao YX Wong CK Li EKM et al Immunomodulatory effects of Lingzhi and San-Miao-San supplementation on patients with rheumatoid arthrtitis Immunopharmacol Immunotoxicol 2006 28(2)197-200
OA COX-2
AutoImmune
disease
Ashwagandha TNFα IL-1β 130 Aggarwah SD Maurya R et al Withania somnifera inhibits ND-kappa B and AP-1 transcription factors in human peripheral blood and synovial fluid mononuclear cells Phytotherapy research 2007 21(10) 905-913
Anti-
inflammatory
Anti-cancer
Anti-viral
Anti-bacterial
Amphoteric
Scutellaria
baicalensis
Reducing NO
expression COX-
2 PGE2 NFk-B
and inflammatory
cytokines IL-1 2
6 12 and TNF-α
131 Kim EH Shim B Kang S et al Anti-inflammatory effects of Scutellaria baicalensis extract via suppression of immune modulators and MAP kinase signaling molecules J Ethnopharmacol 2009 126(2)320-331
QUESTIONS
bull Heidi Kussmann ND FABNO
bull hk365kingmailcom