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Foods For Health:Linking Agriculture to Nutrition and

Health

Bhimu Patil, Director

E-mail: B-patil@tamu.edu

• Current statistics on human diseases

• Questions to be answered– Can chronic diseases be prevented?– What are the evidences?– Phytonutrients: in vitro and in vivo evidences– Linking Food and Agriculture– What are the components of Field to Plate (FTP)

and their role?

• Take home message

Overview

Top 10 causes of death in the U.S

34%

30%

7%

4%

7%

7%

4%3%

2%2%

Heart Disease

Cancer

Stroke

Diabetes

Chronic lower respiratory

Accident

Alzheimers

Influenza

Nephritis

Septicimia

How did we get here????

Economic impact of diseases on U.S economy $$$

0 50 100 150 200 250 300 350

Heart Failure

Stroke

Hypertensive Disease

Coronary Heart Disease

Total Cancer Disease

Diagnosed Diabetes

Total Heart Disease

39.2

73.7

76.6

177.1

201.5

245

316.4

$ Billion

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Global Obesity

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Developing countries: a diabetes riskProjected increase from 2003 to 2025

Source: International diabetes federation

100% increase

DIABETES

0 10 20 30 40 50 60 70 80 90 100

China

India

USA

Brazil

Russian Federation

Mexico

Indonesia

Egypt

Japan

Pakistan

92.3

63.0

24.1

13.4

12.7

10.6

7.6

7.5

7.1

6.6

People With Diabetes (Millions)

FAST FOOD Projected rise in type 2 in India

• Current statistics on human diseases• Questions to be answered

– Can chronic diseases be prevented?

– What are the evidences?– Phytonutrients: in vitro and in vivo evidences– Linking Food and Agriculture

– What are the components of Field to Plate (FTP) and there role?

• Take home message

Overview Top 10 causes of death in the U.S

Heart Disease

Cancer

Stroke

Diabetes

Chronic lower respiratory

Accident

Alzheimers

Influenza

Nephritis

Septicimia

PreventableYES WE

CAN

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Mechanisms of preventing chronic diseases

• Anti-oxidant

• Anti-inflammatory

• Anti-proliferative

• Anti-cancer

• Anti-carcinogenic

• Anti-atherosclerosis

What are the ideal sources to have

optimum levels these activities?

TAKE HOME MESSAGE

Dietary intake and lifestyle has an impact on health and well being

Healthy living 1.1.1

U.S Fruit & Vegetable consumption trends: 2000-2009

0

10

20

30

40

50

(Source: www.cdc.gov)

(%)

Fruit 2> times per day

01020304050

(%)

Vegetables 3> times per day

Trend in homegrown F&V consumption

2020

?Question to be answered

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Bitter Gourd Products Proof of Concept

• In vitro-– Antioxidant

– Enzymatic assays

– Cell culture

• In Vivo

• Clinical trials

Clinical Trials

• 19 out of 21 Studies (+)• Whole food vs Extract vs

Supplement• No effect in two studies

– (-) 2 capsules x3/day (3 Months) - Rand, DB, Controlled (40 Subjects)

– (-) single treatment of Freeze Dried BG juice- Rand, DB, Controlled (ONLY 5 SUBJECTS)

Clinical trials:-Glycated hemoglobin levels (HbA1C)

6.6

6.7

6.8

6.9

7

7.1

7.2

7.3

7.4

7.5

7.6

Base line 8 weeks 16 weeks

%

Placebo

BG Group

Trakoon-osot et al., Journal of Pharmacy Research 2013, 6: 859‐864

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Clinical trial: BG tablet increased insulin level

0

10

20

30

40

50

60

placebo 60mg/kg/day 80mg/kg/day 100mg/kg/day

μlU

/ml

Baseline

15 min

Lim et al., Phillipine Journal of Internal Medicine 2010, 48: 19‐25

Clinical trial: BG tablet decrease plasma glucose levels

155

160

165

170

175

180

185

190

195

200

205

placebo 60mg/kg/day 80mg/kg/day 100mg/kg/day

Baseline

15 min

Lim et al., Phillipine Journal of Internal Medicine 2010, 48: 19‐25

WE ARE ON THE RIGHT PATH At Texas A&M

Explore new varieties & locations for cultivation

Proof of concept: Isolation

Raw material Extraction

Chromatography

Rotovaporation

Characterization

Concentrated extracts

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Bitter Gourd Compounds

•Momordicin (2)•Charantin (2)•Momordicosides ( 24)•Charantiosides (8)•Goyaglycosides (3)•Karavilosides(10)•Karavilagenin (5)•Kuguacin (19)•Kuguaglycoside (8)•Kuguaosides (4)•Charantagenin (5)•Octonorcucurbitacin (4)•Neokuguaglucoside•Charantadiol•Momordicinin•Other Triterpeonids (74)

Flash Separtion

• Faster that traditional open column

• Monitoring capabilities

• Automated fraction collection

• Reduced solvent use

Previous Reports

• 1980-2013 (53 Pubs)– Extraction – Identification – Isolation

• Laborious• High Solvent

consumption• Low yields • Good Review in 2009Lee, Sook Young, et al. "Cucurbitane‐type triterpenoids in Momordica charantia Linn." J Med Plants Res 3.13 (2009): 1264‐1269.

Factors influencing the levels of phytochemicals:Farm to Plate

FARM PLATE

Variety Fertilization Climate

PRE HARVEST FACTORS POST HARVEST FACTORS

Packaging

Maturity

Storage

Processing

Irradiation

TO

Pre harvest factors: Maturity stage Bitter Gourd

Data Adopted from : Pal et al. (2004) Journal of Food Science and Technology, 42: 197‐199.

6466687072747678808284

Green Mature Green Mature

Pusa Hybrid Pusa Vishesh

mg

/100

g

Ascorbic Acid

Pre harvest factors: Variety

Levels of Phenolics Acids

Horax et al., (2010) Journal of Food Science 70: c275‐c280

020406080

100120140160180200

Gallic Acid Catechin ChlorogenicAcid

Total PhenolicAcids

Indian Green Indian White

Chinese Green Chinese White

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Factors influencing the levels of phytochemicals:Farm to Plate

FARM PLATE

Variety Fertilization Climate

PRE HARVEST FACTORS POST HARVEST FACTORS

Packaging

Maturity

Storage

Processing

Irradiation

TO

Effect of Roasting on Antioxidant Activity

0102030405060708090

100

AscorbicAcid

Fruits Leaves Stems Roots

DP

PH

Ra

dic

al

Scav

an

gin

g(%

)

DPPH

Unroasted

Roasted

abb

bb

cd, c

d

b

Choi et al., Food Science and Biotechnology 2012, 28:19‐26

Roasting Effect on Phytonutrient Levels

0

500

1000

1500

2000

2500

Fruits Leaves Stems Roots

Flavan‐3‐ol Derivatives

Unroasted Roasted 0

500

1000

1500

Fruits Leaves Stems Roots

Phenolic Acid Derivatives

Unroasted Roasted

mg

/Kg

mg

/Kg

Post Harvest (other)

• Processing treatments-– Blanching; Bitter masking; fresh; freeze

dried; oven dried; sun dried; fried

– Blanching and Prosweet® masking agent treatments showed absence of momordicoside K and L.

• Storage-– Reduced loss of phytonutrients in cut bitter

gourd stored at 2°C vs whole Wang et al., Journal of Food Processing and Preservation 2007, 31:571‐582

Donya et al., Journal of Agricultural and Food Chemistry 2007, 55: 5827‐5833

Factors influencing the levels of phytochemicals:

FARM PLATE

VarietyFertilization Climate

PRE HARVEST FACTORS POST HARVEST FACTORS

Packaging

Maturity

Storage

Processing

Irradiation

TO

Farm to PlatePlate to FarmBreeding (Bitterness)

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Case study of Curcumin:Traditional uses

Potential Uses of Curcumin Based on Modern technology

Molecular targets of curcumin

Aggarwal, B.B., Surh, Y.J., Shishodia, S., 2006, The Molecular Targets and Therapeutic Uses of Curcumin in Health and Disease, Springer.

Challenges

Rapidly converting to glucuronidation

Piperine is an inhibitor of intestinal and hepatic glucuronidation

Concomitant administration of curcumin with piperine enhanced the bioavailability by 2000% in human volunteers and 154% in rats

Curcumin has shown poor bioavailability due to low absorption and rapid metabolism

Other promising approaches to increase the bioavailability of curcumin in humans include the use of nanoparticles

Dose of 2 g of curcumin consumption, didn’t shown the levels of curcumin in serum

Liposomes, phospholipid complexes, and structural analogues

Increase Public Awareness WE ARE NOW ONE COMMUNITY

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At Texas A&M Road map for HEALTHFUL fruit & vegetable

• Breeding–Traditional

–Biotechnology

• Varietal Selection

• Environment

• Processing

Nutritional Sciences

HealthSciences

• Bioavailability

• Toxicity

• Drug interaction

• Metabolism

• Gut microbial

• Packaging/

Storage

Agricultural Sciences

• Prevention &

Control

• Epidemiological studies

• Population studies

• Public health

LACK COMMUNICATION

BREEDERS

GROWERS

RETAILERHORTICULTURAL SCIENTISTS

CHEMISTS

MEDICALSCIENTISTS

NUTRITIONIST

CONSUMERThe Bridge

Linking Food, Health and Agriculture

Nutrition Bio-Medical

BiotechnologyBreeding and Genetics Plant Physiology

Post-Harvest & Processing

Sports nutrition & Health Sciences

BiochemistryEducation and Economics

VFIC TEAM:

2014

Characterization of Bioactives

VFIC: industry partners

A-W Produce Co., Inc.

Bagley Produce Co.

C&V Supply Inc.

Corey Brothers Inc.

Costa Oro, LLC

Frio Valley Farms

Fruit World

Judy Jackson

Lark Seeds International

Patty Leander

Pan american produce

Super Foods Extracts LLC

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Scientific advisory board

VFIC: Integrated multi-disciplinary research on improving nutritional quality of F&V

Selection Sorting Sample Processing

ExtractionAnalysis

Feed back to breeding program

Sample analysis for clinical trial

Carrot- Breeding to Clinic

High anthocyanin carrot

Breeding for Bioactives

Beta Sweet Maroon Carrot-• Developed through

traditional breeding40 % higher β-carotene

• Improved flavor and texture (high sugar content)

Breeding

Cell culture

Human clinical

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Human intervention study: Breast cancer survivors

0

2

4

6

8

10

Orange Purple

Tota

l pla

sma

caro

teno

ids

(μm

ol/

L)

•3-wk carrot juice intervention,

•8 fluid ounces of carrot juice (mixed with 2 teaspoons of olive oil daily.

Protective level= 4.1879 µmol/LRock et al., 2005

BreedingBreeding

Cell culture

Human clinical

CAX1 Expression increases Ca2+ content in Carrots, Potato, Tomato and Lettuce

Vector   CAX1

1

2

Mol. Breeding, 2004Agric. Food Science, 2005Plant Physiology, 2005Plant Biotechnology, 2009

A

B

Control sCAX1Control sCAX1 Control sCAX1

More Calcium is Absorbed fromCAX1 Carrots than from Controls.

0

5

10

15

20

25

30

35

control CAX1

Carrot type

mg

Ca

abso

rbed

/100

g f

wt.

*

0

100

200

300

400

500

0 1 2 3 4

Conventional OrganicA

scor

bic

aci

d (

µg

/g)

Storage (weeks)

Organic/Conventional

Grapefruit: Ascorbic acid

Organic/Conventional

0

30

60

90

120

150

180

4 11 18 25

Conventional Organic

mg/

100

mL

Hesperidin

Storage (days)

1. Limonin2. Nomilin3. Obacunone4. Deacetylnomilin5. Ichangin6. Deoxylimonin7. Deoxylimonol8. Limonol9. Limonyl acetate10. 7-Obacunyl acetate11. 7-Obacunol12. Ichangensin13. Citusin14. Calamin15. Retrocalamin16. Cyclocalamin17. Methyl-iso-obacunoatedisosphenol18. Methyl deacetylnomilinate19. 6-Keto-7-deacetylnomilol

20. 6-Keto-7 -Nominol21. Isocylocalamin22. 1-(10-19)Abeo-7 -acetoxy-10 -hydroxyisoobacunoic acid 3,10-lactone23. 1-(10-19)Abeo-obacun-9(11)-en-7 0yl acetate24. Deacetylnomilinic acid25. Nomilinic acid26. Iso-Obacunoic acid27. Deoxylimonic acid28. Isolimonic acid29. Epi-iso-obacunoic acid30. 17-Dehydrolimonoic acid A ring lactone31. Calaminic acid32. Retrocalaminic acid33. Cyclocalaminic acid34. Iso-Obacunoic acid disophenol35. Obacunoic acid36. 19-Hydroxydeacetylnomilinic acid37. Limonexic acid

Isolation: Limonoid aglycones

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1. Limonin

2. Nomilin

3. Obacunone

4. Ichangensin

5. Deacetylnomilin

6. Ichangin

7. Calamin

8. Methyl deacetylnomilinate

9. 6-Keto-7-deacetylnomilol

10. Nomilinic acid

11. Deacetylnomilinic acid

12. Obacunoic acid

13. Trans-Obacunoic acid

14. Iso-Obacunoic acid

15. Epi-iso-obacunoic acid

16. Isolimonic acid

17. 19-Hydroxydeacetyl nomilinicacid

17--D-Glucopyranosides of:

Monocarboxylic acids Dicarboxylic acids

Limonoid GlucosidesIsolated/Reported In Citrus

Science of Citrus Health Benefits:In vitro & In vivo

FARM

CHARACTERIZATION

IN VITRO

IN VIVO

CLINICAL

Current status

Several

unknowns……

Pancreatic cancer

Colon cancer

Breast cancer

Limonoids: Cancer

Inhibitory Role of Limonoids in Cancer

Patil et al., J Agric. Food Chem. 57: 10933‐10942

Neuroblastoma

Prostate cancer

Jayaprakasha et al., Food Chemistry, 131:184‐191. 

Kim et al., Eur. J.Pharma. 670: 44‐48

Murthy et al., Food Chem. Tox.49:1616‐1625 Murthy et al.,In vitro Toxicology, 25: 859‐867

Patil et al., Microchem. J., 94:108‐117.

Somasundaram et al., J. App. Glob. Res., 3: 67‐72.

Jayaprakasha et al., J. Agric. Food Chem., 58: 180‐186

Leonardi et al., Exp. Bio. Med., 235:710‐717

Poulose et al, The FASEB Journal 2006. 20:A11‐A12

Gunasekara et al., Nutrition and Cancer 58:117‐121

Basal Diet GFPP IGFPP Naringin Limonin

Tot

al N

umbe

r of

AC

0

50

100

150

200

250

a

b b b

P = 0.02

b

Basal Diet GFPP IGFPP Naringin Limonin

Num

ber

of H

MA

CF

0

2

4

6

8

10

12

14

16

18a

b b

b

P = 0.01

b

GFPP – Grapefruit Pulp PowderIG – Irradiated Grapefruit Pulp Powder

Vanamala et al., 2006. Carcinogenesis 1257‐1264

78

Suppression of colon carcinogenesis by bioactive compounds in grapefruit In vivo 

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Other secondary metabolites from Citrus: Cancer prevention

A Grapefruit Study in Overweight, Healthy Adults: CLINICAL TRIAL

0

50

100

150

200

250

Total Cholesterol (mg/dL)HDL (mg/dL) LDL (mg/dL) Triglycerides

Before GF After GFReduction in total cholesterol after consuming GFJ

Lipid profile changes Educating future generations

72

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Multi-state & Multi-disciplinaryCourses

College Station

Michigan

Weslaco

Indianapolis

Dallas

Acknowledgement

Present research is supported by USDA-CSREES “Designing Foods for Health” through VFIC