Download Pangborn Jon Toronto 2009

COUPLED DYSBIOSIS &
METABOLIC DYSFUNCTIONS IN
AUTISM –POSSIBLE REMEDIES
Jon B. Pangborn, Ph.D., F.A.I.C.,
C.C.N.
TODAY’S ASSIGNMENT:
To explore how impaired methionine-to-taurine
metabolism and reduced methylation, together
with infection, impact autism. We will consider
possible natural and nutritional remedies.
DISCLOSURES
• Father of formerly autistic child – now adult
• Left chemical engineering 29 years ago to investigate
biochemistry and autism
• Past President, Doctor’s Data Laboratory
• Klaire Labs Consultant 1981-1999
• Great Smokies Diagnostic Lab (Genova) 1995-2005
• Cofounder: Defeat Autism Now! 1995
• Coauthor: Autism: Effective Biomedical Treatments
• Consultant: Kirkman Labs since 2000
• Senior Scientific Consultant, ARI
IN PERSPECTIVE
• There are many different autisms:
Primarily Genetic
Mostly acquired
Inherited faults
toxic/infectious stressors
• Considering ASD people as a group, there are cell-wide
and extracellular anomalies.
• Don’t get hung up on one enzyme/protein, one cell
compartment, one gene, one epigenetic process, one
cytokine, one toxicant or infectious agent or any single
theory or model.
TODAY’S FOCUS
• Deficient methylation of lipids adjacent to neuronal
receptors can cause loss of neuronal and network
synchrony. See: Deth, Molecular Origins of Human
Attention Kluwer Acad Pub, 2003.
• Methionine metabolism can be disordered in multiple
ways such that methylation is deficient, oxidant stress
occurs, and an inflammation becomes a persistent,
maladaptive condition.
• Mitochondrial dysfunction is probable when vitamin B12
(as AdoCobalamin) and carnitine are deficient and
maybe when Clostridia dysbiosis occurs.
TODAY’S FOCUS2
• Xenobiotic (e.g. organophosphate) and elemental (e.g.
mercury) toxicants can be contributory.
• Many infectious agents and foreign substances can lead
to oxidant stress when taurine is deficient.
• There are nutritional aids for: methylation, reducing
inflammation, mitochondrial support and alleviating
dysbiosis.
Methionine Transsulfuration to Cysteine and Glutathione
Methionine
THF
5,10CH2THF
MTHFR
1
SAM
MS
2
B12
5-CH3THF
3 Transsulfuration
Pathway
SJJ
(SAM/SAH)
MTase
Cell Methylation
SAH
SAHH
Adenosine
Homocysteine
1 Folate Cycle
2 Methionine Cycle
Methylation
Potential
B6
3
CBS
Cystathionine
Antioxidant
Redox
Cysteine
Potential
GSH GSSG(GSH/GSSG)
B6
CYSTEINE TO TAURINE (main route)
From cystathionine
GSH
Dietary cystine
cysteine
GSH GSCys
cysteine
sulfinic a
dioxygenase
decarboxylase P5P
Stable
Chloroamine
OCltaurine
phagocyte
hypotaurine
oxygenase
CELL MEMBRANE
TRANSCOBALAMIN
CYTOSOL
OHCbl
MeH4folate
methionine
Cbl
MeCbl
GSH
Lysosome
OHCbl
GSCbl
reductase
MS
H4folate
reductase
ATP
PROPIONATE
MMA AdoCbl
SUCCINATE
CARNITINE
mutase
C-P COMPLEX
C.A. cycle
MITOCHONDRION
homocysteine
CARNITINE FORMATION
3 SAM
L-lysine
3 methylations
trimethyl-L-Lysine
CYTOSOL
MITO MEMBRANE
MITO
4 biochemical transformations
L-carnitine
requiring P5P, NAD, O2, ascorbate
and α-ketoglutarate x 2
CARNITINE
CH3
CH3―+N― CH2―CH―CH2―COOH
CH3
OH
ACTIONS OF CARNITINE
• Using a translocase transporter, CARNITINE
escorts fatty acids (as fatty-acyl carnitine)
through the inner mitochondrial membrane.
• Combines with excesses of organic acids
(propionic, isovaleric, 3-methylglutaric, valproic,
others) to accomplish a detoxication function.
REVIEW: FOUR INTERRELATED PARTS
OF BIOCHEMISTRY
1. Methionine → SAM, methylation →→ cysteine with
some homocysteine recycle using methyls provided by
folate to Cbl on MS.
2. Cysteine →→ taurine or cysteine →→ GSH
Taurine scavenges OClduring cellular immune and
defense activities.
GSH neutralizes H2O2 and
detoxifies and makes B12
work.
3. B12 →→GSCbl: cytosolic homocysteine → methionine
and mitochondrial cleanup of propionate via MMA
4. Carnitine, needs methylation for formation, also cleans
up propionate and other excess organic acids.
WHAT’S GOING WRONG?
SO MUCH THAT WE HAVE TO STUDY IT PIECEWISE
METHIONINE → SAM → S-ADENOSYLHOMOCYSTEINE
methylation
Reduced methylation causes varying deficiencies of –
melatonin
choline/TMG/DMG
creatine
methylated catecholamines
carnitine
methylated CG pairs on DNA
methylated fatty acids
Remedies: supplements of melatonin, creatine, L-carnitine,
TMG, DMG. See ARI Publication 34.
CAN A VIRUS IMPAIR METHYLATION IN
NEURONS?
Note: Documentation in rat brain cells of reduced neuronal
receptor phospholipid methylation during paramyxovirus
infection (measles, mumps) – Münzel and Koschel, Proc.
Natl. Acad. Sci. (USA) 79 (1982) 3692-6.
Β-adrenergic receptors affected – loss of catecholaminestimulated methylation
SAH
HOMOCYSTEINE (Hcy)
METHIONINE
ADENOSINE
CYSTATHIONINE
CYSTEINE
1.
Adenosine accumulates, SAH accumulates, SAH/SAM
ratio increases, methylation slows. Adenosine elevated
in ASD, 20% per plasma, 33% per RBCs.
2.
Adenosine Deaminase (ADA) Binding Protein = DPP4
or CD26 is inhibited by: mercury, antimony, lead, organophosphates, streptokinase, casomorphin peptides.
See Püschel et al. Eur J Biochem 126 (1982) 359-65
Vojdani et al., Int.J.Immunopath Pharmacol 16 3
(2003) 189-99
SAH
HOMOCYSTEINE (Hcy)
METHIONINE
ADENOSINE
CYSTATHIONINE
CYSTEINE
3. Genetic variants for ADA, methionine synthase,
formation of cystathionine and methylated folate = risk
factors for ASD.
4. Hcy → Methionine hindered by oxidation of reduced
cobalamin (Co+1)
5. Some Hcy tied up as Hcy thiolactone if PON
(paraoxonase, Hcy thiolactonase) weak – organophosphate toxicity; 40-fold activity variance in humans
6. Functional P5P coenzyme inadequacy
SAH
HOMOCYSTEINE (Hcy)
METHIONINE
ADENOSINE
CYSTATHIONINE
CYSTEINE
REMEDIES FOR POSSIBLE PROBLEMS
• Methylcobalamin and folinic acid
• B6/P5P, Zinc, Magnesium, Melatonin
• Trimethylglycine, Dimethylglycine
• Diet and detoxification (helps ADA Binding Protein –
DPP4)
• Antioxidant nutrients, digestive enzymes
• Not recommended: Methionine, SAM, Cysteine, NAC
CYSTEINE
TAURINE
1.
Cysteine dioxygenase downregulated in autism
(inflammation); see studies by Waring.
2.
Functional P5P coenzyme inadequacy
3.
Urinary loss of taurine due to beta-aminoaciduria,
mostly elevated beta-alanine –
Bacterial dysbiosis
Anserine and carnosine
Pyrimidine disorder, PDD, see studies by Page
CYSTEINE
TAURINE
REMEDIES FOR POSSIBLE PROBLEMS
• Antiinflammatory meds, digestive aids
• B6/P5P, zinc, magnesium, melatonin
• Antioxidant nutrients
• Taurine, 100-250 mg/day with breakfast
RELEASE OF OXIDANTS BY PHAGOCYTIC
NEUTROPHILS (PMN LEUKOCYTES) WHEN TAURINE
IS INADEQUATE
Phagocyte – a cell that ingests microorganisms, foreign
particles, other cells, and kills, digests or decomposes
them.
Oxidant killing is a normal, body-defense mechanism:
Respiratory Burst
O2-, H2O2, OClTaurine limits collateral damage by scavenging OClSee Dinauer et al., Chapt 189 in Scriver et al., Eds., The
Metabolic and Molecular Bases of Inherited Disease, 8th
Ed.
SEE: DISTURBANCES OF AMINO ACID METABOLISM:
CLINICAL CHEMISTRY AND DIAGNOSIS, Hans J.
Bremer, et al., Urban & Schwarzenburg, 1981 p225
Table B24. Free amino acids in human polymorphonuclear
leukocytes (Houpert et al. 1976)
Amino Acid
nmoles/million cells
Taurine
15.47
Aspartic acid
1.52
Alanine
1.51
Lysine
1.36
Glutamic acid
1.29
Astrocytes: Taurine also the most abundant AA. See
Urenjak et al. J.Neurosci 13, 3 (1993) 981-89
WHAT PROVOKES PHAGOCYTIC ACTIVITY
AND OXIDATIVE PROCESSES?
• Bacteria (staph, strep, Salmonella, Enterobacter,
Proteus, Klebsiella, Microbacteria, others)
• Yeasts (Candida, Aspergillus)
• Wheat germ agglutinin, substances that “clump” cells
• Fine particles, organic & inorganic, engine & powerplant
exhausts
• Fluorides
• Some viruses (naked virus w/o lipid envelopes – rhino,
papilloma, rotoviruses)
CLOSTRIDIA OVERGROWTH
• Anaerobic organism; can’t assay by standard culture,
often identified by chemical markers: propionic acid,
phenyl & hydroxyphenyl propionates, indole propionate,
phenol, para-cresol, acetic acid, acetates, phenyllactate.
- See Elsden et al. Arch.Microbiol 107,3 (1976) 283-8
• Many types: C.difficile, propionicum, tetani (credit to
Ellen Bolte, 1996), sporogenes, botulin, and many more.
• Propionic acid and propionates induce autistic behavior
in rats – See work of Derrick MacFabe.
CLOSTRIDIUM DIFFICILE: ANTIBIOTIC RESISTANT
STRAIN
• First public notice: Warrey & Pipin - Lancet 2005; Cloud
& Kelly – Curr Opinion Gastroenterol 23,1 (207) 4-9;
Baines & O’Connor – J.Microb.Chemother 62,5 (2003)
1078-85; Spigaglia et al. PMID 18480338; Saxon et al.
PMID 18710908
• Vancomycin sends it to spore form; later it comes back
with a vengeance
• Ciprofloxacin, Levofloxacin and 20 more antibiotics (esp.
fluoroquinolones) have no lasting effect. “What almost
kills it, makes it stronger.”
NATURAL ANTIBIOTIC FUNCTION IS A
TWO-PRONGED ATTACK
A.
B.
Toxic killing agents, plus
Agent that disables organisms’ defenses
See: Proc.Natl.Acad.Sci (USA) 97 (2000) 1433
Drs. Lewis (Tufts) and Stermitz (Col. State U)
Studied Barberry: berberine + 5’methoxyhydnocarpin
WHAT HERBALS HAVE NATURAL ANTIBIOTIC
ACTION AGAINST CLOSTRIDIA?
• Thyme, oregano, cumin (at low levels). See Dorman &
Dean, J. Appl.Microbiol. 88 (2000) 308-16
• Curcumin (turmeric) provides antioxidant protection,
lowers cytokines (inc. TNF-α), regulates COX-2 and
downregulates NF-Kappa B.
• See –
Chainani-Wu J.Alt.Complement Med 9 (1) 2003 161-8
Bundy, Walker et al. Prac Nutr Soc 63 2004 126A
Guo, Kuo et al. Biochem Pharmacol 680 2004 51-61
Menon & Sudheer Adv Exp Med Biol 575 2007 105-25
WHAT PROBIOTICS REDUCE OR HELP TO
KILL C. DIFFICILE?
Saccharomyces boulardii is effective for C. difficile
disease. S. boulardii and Lactobacilus rhamnosis both
reduce antibiotic-associated diarrhea.
See: McFarland LV “Meta-analysis of probiotics for the
prevention of antibiotic-associated diarrhea and the
teatment of Clostridium difficile disease” Am J
Gastroenterol. 2006 Apr: 101 (4) 812-22
Segarra-Newnham M “Probiotics for C-difficileassociated diarrhea: focus on L. rhamnosis GG and S.
boulardii” Ann.Pharmacother 2007 Jul; 41 (7): 1284-7
WHAT PROBIOTICS REDUCE OR HELP TO
KILL C. DIFFICILE?
Lactobacillus plantarum administration to antibiotic-treated
patients reduces colonization of C. difficile.
See –
Plummer S, Weaver MA et al. “C. difficile pilot study: effects
of probiotic supplementation on the incidence of C.
difficile diarrhea” Int Microbiol. 2004 Mar; 7 (1): 59-62
Klarin B, Wulft M et al. “L. plantarum 299v reduces
colonization of C. difficile in critically ill patients treated
with antibiotics” Acta Anaesthesiol Scand. 2008 Sep; 52
(8): 1096-102
SHORT HISTORY OF MEDICINE
Problem: I have an earache….
2000 B.C.
1000 A.D.
1850 A.D.
1940 A.D.
1985 A.D.
1997 A.D.
Here, eat this root.
That root is heathen. Here, say this prayer.
That prayer is superstition. Here, drink this
potion.
That potion is snake oil. Here, swallow this
pill.
That pill is ineffective. Here, take this
antibiotic.
That antibiotic ruins your own immunity. Here,
eat this root.