COUPLED DYSBIOSIS

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COUPLED DYSBIOSIS METABOLIC DYSFUNCTIONS IN
AUTISM POSSIBLE REMEDIES

• Jon B. Pangborn, Ph.D., F.A.I.C., C.C.N.

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TODAYS 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.

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DISCLOSURES

• Father of formerly autistic child now adult
• Left chemical engineering 29 years ago to
  investigate biochemistry and autism
• Past President, Doctors 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

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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.
• Dont 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.

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TODAYS 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.

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TODAYS 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.

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Methionine Transsulfuration to Cysteine and
Glutathione
Methionine
Methylation Potential (SAM/SAH)
THF
SAM
MTase
2
1
5,10-CH2THF
Cell Methylation
MS
SAH
B12
MTHFR
5-CH3THF
SAHH
Adenosine
Homocysteine
1
Folate Cycle Methionine Cycle Transsulfuration
Pathway SJJ
CBS
B6
Cystathionine
2
Antioxidant Redox Potential (GSH/GSSG)
B6
3
Cysteine
3
GSH GSSG
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CYSTEINE TO TAURINE (main route)

• From cystathionine GSH
• 
  cysteine
• Dietary cystine cysteine
  sulfinic a
• GSH GSCys
  dioxygenase
• 
  decarboxylase P5P
• Stable OCl-
• Chloroamine taurine
  hypotaurine
• phagocyte
  oxygenase

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• CELL MEMBRANE
• TRANSCOBALAMIN
• OHCbl
  CYTOSOL
• 
  MeH4folate methionine
• GSH
• Lysosome OHCbl GSCbl
  Cbl MeCbl
• 
  reductase MS
• 
  H4folate
  homocysteine
  
• 
  
• 
  
  
• reductase ATP
• PROPIONATE
• MMA
  AdoCbl SUCCINATE
• CARNITINE mutase
• C-P COMPLEX
  C.A. cycle
• 
  MITOCHONDRION

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CARNITINE FORMATION

• 3 SAM
• L-lysine
  trimethyl-L-Lysine
• 3 methylations
  CYTOSOL
• MITO MEMBRANE
• 
  MITO
• 4
  biochemical transformations
• L-carnitine
  requiring P5P, NAD, O2, ascorbate
  
  and a-ketoglutarate x 2

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CARNITINE

• CH3
• CH3?N? CH2?CH?CH2?COOH
• CH3 OH

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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.

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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 OCl- GSH neutralizes H2O2
  and
• during cellular immune and detoxifies and
  makes B12
• defense activities. 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.

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WHATS 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.

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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.
• ?-adrenergic receptors affected loss of
  catecholamine-stimulated methylation

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• SAH HOMOCYSTEINE (Hcy) METHIONINE
• ADENOSINE
• CYSTATHIONINE
  CYSTEINE
• Adenosine accumulates, SAH accumulates, SAH/SAM
  ratio increases, methylation slows. Adenosine
  elevated in ASD, 20 per plasma, 33 per RBCs.
• Adenosine Deaminase (ADA) Binding Protein DPP4
  or CD26 is inhibited by mercury, antimony, lead,
  orga-nophosphates, 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

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• 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 (Co1)
• 5. Some Hcy tied up as Hcy thiolactone if PON
  (paraoxonase, Hcy thiolactonase) weak
  organophos-phate toxicity 40-fold activity
  variance in humans
• 6. Functional P5P coenzyme inadequacy

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• 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

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• CYSTEINE TAURINE
• Cysteine dioxygenase downregulated in autism
  (inflammation) see studies by Waring.
• Functional P5P coenzyme inadequacy
• Urinary loss of taurine due to beta-aminoaciduria,
  mostly elevated beta-alanine
• Bacterial dysbiosis
• Anserine and carnosine
• Pyrimidine disorder, PDD, see studies by Page

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• CYSTEINE TAURINE
• REMEDIES FOR POSSIBLE PROBLEMS
• Antiinflammatory meds, digestive aids
• B6/P5P, zinc, magnesium, melatonin
• Antioxidant nutrients
• Taurine, 100-250 mg/day with breakfast

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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, OCl-
• Taurine limits collateral damage by scavenging
  OCl-
• See Dinauer et al., Chapt 189 in Scriver et al.,
  Eds., The Metabolic and Molecular Bases of
  Inherited Disease, 8th Ed.

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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

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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)

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CLOSTRIDIA OVERGROWTH

• Anaerobic organism cant 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.

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CLOSTRIDIUM DIFFICILE ANTIBIOTIC RESISTANT STRAIN

• First public notice Warrey Pipin - Lancet
  2005 Cloud Kelly Curr Opinion Gastroenterol
  23,1 (207) 4-9 Baines OConnor
  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.

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NATURAL ANTIBIOTIC FUNCTION IS A TWO-PRONGED
ATTACK

• 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 5methoxyhydnocarpin
  

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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-a),
  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

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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-difficile-associated diarrhea focus on L.
  rhamnosis GG and S. boulardii Ann.Pharmacother
  2007 Jul 41 (7) 1284-7

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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

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SHORT HISTORY OF MEDICINE

• Problem I have an earache.
• 2000 B.C. Here, eat this root.
• 1000 A.D. That root is heathen. Here, say this
  prayer.
• 1850 A.D. That prayer is superstition. Here,
  drink this potion.
• 1940 A.D. That potion is snake oil. Here, swallow
  this pill.
• 1985 A.D. That pill is ineffective. Here, take
  this antibiotic.
• 1997 A.D. That antibiotic ruins your own
  immunity. Here, eat this root.