Urinary pyrrole (Mauve Factor): marker for oxidative stress in behavioral disorders
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Transcript of Urinary pyrrole (Mauve Factor): marker for oxidative stress in behavioral disorders
Urinary pyrrole (Mauve Factor): marker for oxidative stress in
behavioral disorders
Woody McGinnis [email protected]
Seattle, 6 November 2004
CH3 C2H5
The Mauve Factor
OHHPL (hydroxyhemopyrrolin-2-one)
OHHPL (Mauve Factor)
• In human urine, blood and CSF
• Mistakenly identified as kryptopyrrole, a persistent erroneous term
• Chemically similar to kryptopyrrole, which can be used for OHHPL assay
CH3 C2H5 C2H5 CH3
HP KP
CH3 C2H5 C2H5 CH3
OHHPL OHKPL
Mauve history• Discovered in urine in 1957
• Named for lilac-colored appearance on paper chromatograms developed
with Erhlich's reagent
• Labile and elusive
• Abram Hoffer is the father of Mauve
Mauve Hall of FameHofferIrvineOsmondPfeifferSohlerCutlerO'Reilly
GrahamRiordanJacksonWalshAudhya
Europe
Hoffer J Neuropscyh 1961
• Qualitative Mauve assay
• All normals mauve-negative
• 27/39 early schizophrenics positive
• All 7 who recovered on niacinamide converted to negative
Hoffer 1961
• Relapses associated with reappearance of Mauve
• Apparent role in other behaviors: ETOH, depression.
• A "mentally retarded" mauve-positive child responded dramatically to
niacinamide
Hoffer and Mauve • Heat and light sensitive
• Relatives should be tested• Preventive potential
• 10/14 criminal / deviant positives• Report on 740 patients in 1966• All recovered schizophrenics
negative, unrecovered 50% positive
O'Reilly 1965
• Report on 850 behavioral patients
• 25% of "disturbed children" mauve-positive, vs 12% of well children
• First documented observation of Mauve association with stress
Mauve in schizophrenia
• Hoffer 1961, 1963, 1966
• Yutwiller 1962
• O'Reilly 1965
• Sohler 1967 x 2
High-Mauve and behavior
• Down syndrome 70%
• Schizophrenia 40-70%
• Autism 50%
• ADHD 30%
• ETOH 20-80%
Carl Pfeiffer 1972• "Sara" 15 y.o. with four years of unreality
spells, insomnia, seizures, attempted suicides, knee problems; quite well on
B6 1000 mg, Zn 160 mg, Mn 8 mg.
• Signs, symptoms, and clinical response imply high B6 / zinc need in high-Mauves.
• B6 and zinc quickly recognized by clinicians as main-stay treatment.
Mauve levels• Clinicians: behavioral symptoms in
individuals correlate with level• Irvine 1972: likelihood of depressive
reactions correlate with level• Cutler 1974: B6 dose needed to
normalize Mauve proportional to level• McCabe 1983: Mauve can be normalized with high-dose B6 only
Pfeiffer 1983• Symptoms may improve in 24 hours,
usually within 1 week
• May need months for full recovery
• Relapse within days or weeks if no nutrients
• Changing needs
OHHPL Levels and B6 (10mg/kg/day) + Zn (25mg) + Mg (400mg) in Autism
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Pfeiffer correlates
• Nail spots• Stretch marks• Pale skin• Poor tanning• Knees and joints• Constipation
• Dream recall• Morning nausea• Light and sound• Odor intolerance• Migraines• Stitch-in-side
Walsh• Low stress
tolerance• Anxious, overly
pessimistic• Explosive anger• Hyperactivity
Kruesi• Social
withdrawal• Emotionally
labile• Loss of appetite• Fatiguability
Mauve and stress
• Mauve is associated with stress, including, emotional stress.
Audhya 1992. Cold-immersion increased Mauve <1 hour
• The correlation is well-documented over decades
Mauve and stressO'Reilly 1965Sohler 1971
Pfeiffer "stress-dosing" 1973Ward says across all diagnoses 1975
Hippchem 1978McCabe 1983
Jaffe and Kruesi 1992
Non-behavioral Mauve• Acute Intermittent Porphyria
• Cutler 1974: High-mauve obesity and abnormal glucose tolerance
• Hoffer 1966: 33/99 Cancer patients, 7/8 lung cancer patients
• Riordan and Jackson: 43% of general medical patients: arthritis, chronic fatigue,
heart disease, hypertension, irritable bowel, migraine. Range 20-40 mcg%.
Mauve bumps in the road• U Michigan 1962: no pathological
importance in schizophrenia because found 34% Mauve-negative.
• U Cal 1969: their "simplified" assay produced phenothiazine false-positive
• India 1971: No Mauve in 120 psych patients--used HCl, 24° collection.
More bumps in the road..• Am J Psychiatr 1978: "Pyroluria a
poor marker in chronic schizophrenia" (based on 2/9 Mauve-positives)
• J Nutr 1979: "..urinary kryptopyrrole..proved invalid as screening test for vitamin dependent disorders.." (based on 6/20 Mauve-positives, all borderline. And no zinc.)
Bumps• U Cal 1975: "Non-occurrence of
kryptopyrrole and hemopyrrole in urine of schizophrenics by GC-MS"
• UC Berkeley 1978: GC/MS shows no kryptopyrrole or hemopyrrole in schizophrenics or controls
• Irvine 1977-78 confirms Mauve is OHHPL by synthesis.
Bumps..• Irvine, Nature 1969: Mauve identified
unequivocally as kryptopyrrole
• Irvine, in landmark Orthomolecular Psychiatry, "Mauve is kryptopyrrole"
• Irvine 1974: lactam of kryptopyrrole is the "identity of the natural kryptopyrrole"
Was Pfeiffer right about Mauve and low zinc?
Walsh. 1148 ADHD patients:
Plasma Zn vs colorimetric MauveStrong negative correlation
0.974 significance (F test)
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OHHPL
RB
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OHHPL vs.RBC Zinc
Correlation Coefficient -.985
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Mauve
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inc
Correlation Coefficient -.743
Mauve vsWBC Zinc
Mauve as clinical tool• Careful specimen collection
• Mild 20-30, moderate 30-40, severe over 40 mcg%
• Elevations imply zinc and B6 need
• Titrate nutrients to suppress Mauve
• Individualize adjunctive nutrients
Mauve is OHHPL• Graham, Univ Glasgow 1978 quantified
normal range by GLC• Audhya 1994-present: commercial OHHPL
by HPLC/MS and synthetic standard.• Strong logical imperative to cease
"kryptopyrrole" terminology
OHHPL facts• Irvine 1977: levels correlate with
emotional withdrawal, motor retardation, blocked affect and severe depression; IP to rats: ptosis, locomotor aberration, hypothermia
• Cutler 1990: IP to mice increased backward locomotion and head-twitching (as with psychotomimetics)
OHHPL facts• Photo, heat, and acid-labile• Urinary half-life 10-12 hours
• Nearly 100% urinary clearance intact after IP administration
• Daily excretion up to 1 mg • Urine, Blood, CSF; animal brain
• Graham: similarity to kainic acid and pyroglutamate suggests excitoxicity
Why do they both work?
Niacinamide (B3)OR
Vitamin B6 (P5P) and Zinc
Thinking points• B3, Zn and B6 are anti-oxidant
• Strong stress / Mauve association• Emotional stress clearly causes
oxidative stress• The behavioral and somatic high-
Mauve disorders feature high oxidative stress
High Oxidative Biomarkers• Down Syndrome• Schizophrenia
• Autism• ADHD
• Emotional Stress• Cancer and Inflammatory Disease
• Hyperglycemia
Zinc is anti-oxidant• Shields -SH groups
• Blocks lipid peroxidation and PLA2
• Induces metallothionein
• Constituent of SOD
• Maintains vitamin A
• Deficiency increases intestinal NO˙
Zinc deficiency increases oxidative stress
• Lower glutathione, vitamin E, GST, GSHPx and SOD
• Increased reactive species and lipid peroxides in tissue, membranes and
mitochondria
Oxidants mobilize zinc
• Oxidants release complexed zinc from zinc-binding proteins, including
metallothionein and albumin
• It is likely--but unproven--that zinc retention is reduced in direct
relationship to oxidative stress
Oxidative stress
Low zinc
B6 is anti-oxidant
• P5P for Glutathione, Metallothionein, CoQ10 and Heme synthesis
• With Zn, cofactor for GAD
• P5P protects vulnerable lysinyl groups, as in GSHPx
Marginal B6 deficiency:
Lowers GSHPx
Lowers glutathione reductase
Promotes mitochondrial decay
Raises lipid peroxide levels
B6 and oxidative stress• Binding of P5P-dependent enzymes
is subject to carbonyl inhibition
• Binding of key P5P-dependent enzymes such as GAD impaired by
oxidants generally
• OH˙ and 1O2 attack B6 vitamers
B6 and Mauve• B6 levels are normal
• Pfeiffer alluded to lower P5P and EGOT activity in high-Mauves
• Lower zinc may impair B6 activation• Oxidative stress affects activation of
B6 and binding of B6-dependent enzymes
B3 is anti-oxidant
• NADPH for reduction of glutathione
• Potent free-radical quencher: protects both lipids and proteins from oxidation
• Blocks NO˙ neurotoxicity
• High tissue levels: better lipoxidation prevention than ascorbate
B3 is anti-oxidant
• Niacin antagonists increase lipoxidation
• Low B3 decreases MT and increases apoptosis in brain cells
• Neuroprotective in experimental mitochondrial toxicity
Reciprocal relationships
Oxidative stress
Poor energetics Excitotoxicty
Require heme
• Cystathionine synthase
• Catalase• Heme-hemopexin
for MT translation• Pyrrolase
• Guanylate cyclase
• Cytochromes• Sulfite
reductase• NOS
OHHPL and heme• Durko 1970. Oxidized kryptopyrrole
binds heme in vitro
• Graham 1979. Microsomal levels 48 hours after IP OHHP to rats:
Heme down 42% Cytochrome p450 down 50%
Regulatory heme
• Sustains zinc, vitamin A and melatonin levels
• Differentiation, response to growth factors and resistance to viruses
• Levels lowered by toxins: gasoline, benzene, arsenic and cadmium
Low Heme is pro-oxidant• Ames 2002. Experimental heme
depression in cultured brain cells:
Decreased Complex IV 50% reduction in intracellular Zn
Increased intracellular FeIncreased NOS (x 7), NO2 and NO3
Heme depression
• Neuroblastoma cells fail to differentiate, axons degenerate
• Astrocytoma cells fail to proliferate
• Heme synthesis itself depends on sufficient B6 and Zn
HypothesisMauve may be a significant contributer
to oxidative stress. So, it may be a good biomarker for oxidative stress.
• Heme depression, with impaired energetics, zinc, and detox
• Mauve excitotoxicity
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Mauve vs. RBC Glutathione
CorrelationCoefficient -.408
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Mauve vs. GST
CorrelationCoefficient -.65087
p<.02
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OHHPL
Bio
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OHHPL vs. Biotin
CorrelationCoefficient -.973
A second hypothesis• Mauve may form as a result of
oxidative stress.• Sequential lipid oxidation, protein
adduction, oxidative side-chain scission and dissociation.
• Models: Isolevuglandin pyrroles and urinary pyrroles from hexane
Proposed mechanism
• Sequential lipid oxidation, protein adduction, oxidative side-chain
scission and dissociation.
• Models: Isolevuglandin pyrroles and urinary pyrroles from hexane
Oxidative by-product?Some consistencies:
• In the high-Mauve schizophrenics, membrane arachidonate, presumed substrate, is lower.
• Formation of the pyrrolic adduct occurs at lysinyl groups, so may impair pyridoxal kinase and the B6-dependent enzymes.
More Mauve research!• Controlled clinical trials
• Other therapeutic and toxic modulators
• Pro-oxidant and excitotoxic properties
• Prove origin
• Genetic predisposers
The Oxidative Stress in Autism Study
Principle Investigator: William WalshNIH Author: Robert Salomon
Collaborators: Bruce Ames, Allen Lewis, George Perry, Domenico
Pratico, Aristo VojdaniSponsors: Alexander and Bo MacInnis