Transcript Clostridia metabolites inhibit human dopamine

Clostridia metabolites inhibit human
dopamine-beta-hydroxylase,
increasing toxic dopamine
metabolites and severe oxidative
stress in autism
William Shaw PhD
The Great Plains Laboratory,Inc
www.gpl4u.com
Dialogues Clin Neurosci. 2011;13:55-62.
Autism Insights 2010:2 1–11
William Shaw
Nutritional Neuroscience 2010 Vol 13 No 3: 1-10
Structure of 3-(3-hydroxyphenyl)3-hydroxypropionic acid
5
6
3
2
1
1 CHOHCH2COOH
4
Hydroxyl group
3
HO
Phenyl group
2
Propionic acid
group
Data from previous Shaw article
Journal of Biological Chemistry 225:269-278,1957
• “It was observed that mentally ill patients, in
general, seem to excrete much larger amounts
of HPHPA than do most normal people.”
• Most patients with mental retardation excrete
very low amounts of HPHPA.
Increased Urinary Excretion of Analogs of Krebs Cycle
Metabolites and Arabinose in Two Brothers with Autistic Features.
W Shaw. Clin Chem 41:1094-1104, 1995.
HPHPA
Child psychosis during hospitalization
(simulated from memory)
HPHPA
HPHPA
Clostridia species that
produce HPHPA precursors
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•
•
•
•
•
•
C. difficile-pseudomembranous colitis
C. sporogenes
C. botulinum-food poisoning
C. mangenoti
C. ghoni
C. bifermentans
C. caloritolerans
Figure 2. Distribution of values for HPHPA Clostridia
metabolite in urine samples of male infants, control
boys, and boys with autism.
2000.00
3-(3-hydroxyphenyl)-3-hydroxypropionic acid
(MMOL/MOL CREATININE)
1800.00
1600.00
1400.00
1200.00
1000.00
N = 211
800.00
600.00
N = 30
400.00
200.00
N = 14
0.00
CONTROL
Control
MALE
INFANTS
infants
Control
CONTROL
MALE
boys CHILDREN
2-13 years
2 - 13 YRS
Autistic
AUTISTIC
BoysMALES
13 YRS
2-132 -years
Main brain neurotransmitters
Norepinephrine and dopamine
Catecholamine Synthesis
Mainly adrenal
gland
Dopamine
Norepinephrine
Catabolism of Dopamine
Organic acid test
Norepinephrine Catabolism
Organic acid test
Endoscopy of colon of patient with
mild Clostridium difficile overgrowth
Endoscopy of colon of patient with severe
Clostridium difficile overgrowthpseudomembranous colitis
Formation of p-cresol from
tyrosine by Clostridia bacteria
PST
tyrosine
4-hydroxyphenylpyruvic
4-hydroxyphenylacetic
p-cresol
14728–14733 PNAS August 25, 2009 vol. 106 no. 34
p-cresol sulfate
phenylalanine
phenylalanine
hydroxylase
CH2CHCOOH
CH2CHCOOH
CH2CHCOOH
NH2
NH2
NH2
HO
HO
phenylpropionic
acid
microbial
NH2
microbial
tyrosine analog
tyrosine
hydroxylase
CH2CH2COOH
HO
CH2COCOOH
4-hydroxyphenylpyruvic acid
NH2
HO
HO
human beta
oxidation enzymes
CH3
HO
dopamine
COOH
4-cresol
HVA
epinephrine
3-(3-hydroxyphenyl)
-3-hydroxypropionic
acid (HPHPA)
3,4-dihydroxyphenyl
Alanine (DOPA)
norepinephrine
3-hydroxyphenylpropionic acid
human beta-oxidation
CHOHCH2COOH
CH2CHCOOH
HO
CH2CH2COOH
microbial
tyrosine
HO
NH2
VMA
glycine
HO
3-hydroxybenzoic
acid
3-hydroxybenzoylglycine
(3-hydroxyhippuric)
Critical effect of intestinal bacteria on brain
neurotransmitters
Organic acid test
Organic acid
test
Organic acid test
Brain
Intestine
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* * *******
* ** *** *
*****
*******
* ** *
*
*
Body
* *
*
* Kidney
*
*
Blood vessels
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* B* *
*
* B*
*
*
**
**
* ***
*
*
*
*
=Clostridia
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*
bacteria
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**
*
* = products
Urine cup
*
0f Clostridia
*
B= bacteria
****
(beneficial)
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*** **
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Effect of HPHPA on brain
neurotransmitters
(Dopamine)
Clostridia
HPHPA
Mmol per
Mol
creatinine
HVA
Mmol per
Mol
creatinine
Date
Effect of HPHPA on brain
neurotransmitters
30
Dopamine
metabolite
HVA
Mmol per
Mol
creatinine
25
7
VMA (norepi)
6
5
20
4
15
3
10
HVA (dopamine)
2
5
1
0
0
11/18
2/26
6/6
Date
9/14
12/23
4/1
Norepinephrine
Metabolite
VMA
Mmol per mol
creatinine
Effect of HPHPA on brain
neurotransmitters
5000
8
4500
7
4000
HPHPA
Mmol per
Mol
creatinine
6
3500
5
3000
2500
4
2000
3
1500
2
1000
1
500
0
11/18
0
2/26
6/6
Date
9/14
12/23
4/1
HVA/VMA
Indicator of
Dopamine to
Norepinephrine
ratio
Linan Chen, et al(2008) Unregulated cytosolic dopamine
causes neurodegeneration associated with oxidative stress in
mice. J. Neurosci. 28, 425–433
• Dopamine is a very reactive molecule compared with other
neurotransmitters, and dopamine degradation naturally
produces oxidative species.
• More than 90% of dopamine in dopamine neurons is stored
in abundant terminal vesicles and is protected from
degradation.
• However, a small fraction of dopamine is cytosolic, and it is
the major source of dopamine metabolism and presumed
toxicity.
• Cytosolic dopamine undergoes degradation to form 3,4dihydroxyphenylacetic acid (DOPAC) and HVA as well as
hydrogen peroxide via the monoamine oxidase pathway.
Figure 1. Toxicity of excess dopamine
Homovanillic acid
(HVA)
Dihydroxyphenylacetic
(DOPAC)
Linan Chen, et al(2008) Unregulated cytosolic dopamine
causes neurodegeneration associated with oxidative stress in
mice. J. Neurosci. 28, 425–433
• Alternatively, dopamine undergoes oxidation
to form superoxide, hydrogen peroxide, and oquinone or reacts with cysteine residues on
glutathione or proteins to form cysteinyldopamine and cysteinyl-DOPAC conjugates.
• These biochemical abnormalities caused by
excess dopamine may cause severe
neurodegeneration of neural pathways that
utilize dopamine as a neurotransmitter.
Prevalence of Clostridium difficile in the gastrointestinal
tract of hospitalized children under two years of age. Med
Dosw Mikrobiol; 2010;62(1):77-84 (Poland)
• 178 fecal samples of children aged 2 months to 2
years,hospitalized in 2003-2006 were examined
for the presence of toxin A/B of C. difficile.
• Toxigenicity of strains was confirmed using PCR.
• Susceptibility to antimicrobials was determined.
• The percentage of children infected with C.
difficile was 68.6%.
• All strains were susceptible to Vancomycin and
metronidazole(Flagyl)
Summary
• Clostridia metabolites HPHPA, 4-cresol, and 4hydroxyphenylacetic are toxic because they inhibit
key enzyme that converts dopamine to
norepinephrine
• Toxic dopamine metabolites use up glutathione and
person is much more susceptible to other toxic
chemicals like mercury, pesticides, antibacterial
soaps
Acta psychiatr belg 80:249-265,1980
Main dopamine
metabolite HVA
mmol/mol
creatinine
p<0.01
B. Garreau et al. Disturbances in dopamine metabolism
in autistic children: results of clinical tests and urinary
dosages of homovanillic acid (HVA). Acta psychiatr belg
80:249-265,1980.
• Dopamine metabolite (homovanillic acid or HVA) in urine
was more elevated in children with autism than in PDD
and the severity of symptoms was related to the
concentration of HVA
• High dopamine production in brain is associated with
repetitive, stereotypical, obsessive, compulsive behaviors
• The effects of norepinephrine are alertness and arousal,
and influences on the reward system. Norepinephrine is
important for the exploratory behavior essential for
learning relations between sensory input, decision
processing, motor output, and behavioral feedback.
Organic Acids Test: Great Plains Lab
Organic Acids Test: Great Plains Lab
Before Treatment
HVA/VMA= 2.58 (Excess dopamine)
After Treatment
HVA/VMA= 1.38
Factors involved in autism, Clostridia
toxicity measured in urine organic acid
test
• HVA-(homovanillic acid)-Major metabolite of
dopamine, a major brain neurotransmitter
associated with abnormal autistic behavior when it is
elevated
• VMA-(vanillylmandelic acid)- Major metabolite of
norepinephrine- important for the exploratory
behavior essential for learning relations between
sensory input, decision processing, motor output,
and behavioral feedback.
Factors involved in autism, Clostridia
toxicity measured in urine organic acid
test
• 4-hydroxyphenylacetic- Clostridia metabolite that is
a phenol that is detoxified by phenol sulfotransferase (PST), leading to increased susceptibility
to acetaminophen toxicity. 4-Cresol and HPHPA are
also PST inhibitors.
• HVA /VMA ratio –indicates whether there is a
healthy balance in the brain between norepinephrine
and dopamine
Factors involved in autism, Clostridia
toxicity measured in urine organic acid
test
• Pyroglutamic acid- high values indicate deficiency of
glutathione, a major cause of dopamine toxicity and
increased susceptibility to most environmental
chemicals
• 4-Cresol- Major metabolite of Clostridium difficileblocks dopamine beta hydroxylase leading to excess
dopamine and abnormal behavior
• HPHPA-Major metabolite of multiple Clostridia
species- blocks dopamine beta hydroxylase leading
to excess dopamine and abnormal behavior
Clinical usefulness of Clostridia
treatments
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Schizophrenia
Psychosis
Depression
Chronic fatigue
Tics, Tourette’s
Autism
• ADD, ADHD
• Obsessive compulsive
disorder (OCD)
• Seizure disorders
• Gastrointestinal
disorders, diarrhea,
constipation,Crohn’s
disease,colitis
Effects of 3-hydroxyphenylalanine
in rats
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Headweaving
Predominantly backward walking
Wet dog shakes
Hyperactivity
Hyper-reactivity
Dyck LE, Kazakoff CW, Dourish CT. The role of catecholamines, 5-hydroxytryptamine and m-tyramine in
The behavioural effects of m-tyrosine in the rat. European Journal of Pharmacology 1982; 84(3-4): 139149
Two months of nystatin and Lactobacillus
acidophilus GG therapy in a child with autism
Candida
krusei
stool
Yeast Lacto- Clostridia
tartaric bacillus HPHPA
urine* stool
urine*
Before
4+
993
0
3265
After
0
1
4+
174
0-1+
0-15
3+ - 4+
normal
range
0-150*
mmol/mol
creatinine
Effect of anti-Clostridia therapy
on urine excretion of HPHPA* in
young woman with acute
psychosis-auditory
hallucinations
During acute
psychosis
After treatment
( depressed but no
psychosis)
*mmol/mol creatinine
patient
7489
normals
0-150
673
0-150
Effect of anti-Clostridia therapy on urine
excretion of HPHPA* in woman with
depression and chronic fatigue
Before treatment
After treatment
*mmol/mol
creatinine
patient
1444
normals
0-150
13
0-150
Treatments for Clostridia bacteria
• Vancomycin-oral-not intravenous-5-10 mg/Kg/day div into 3
doses
• Flagyl (metronidazole)-30 mg/Kg/day div into 3 doses-10 days
• Lactobacillus acidophilus GG-10 -100 billion per day
• Alternate name Lactobacillus rhamnosus
• (Only bacteria probiotic patented for use in control of
Clostridia)-Culturelle-VSL #3
• Saccharomyces boulardi (yeast)
• Micellized or IV glutathione or n-acetylcysteine to increase
brain glutathione and reduce neurotoxic dopamine
metabolites
• High protein diet (phenylalanine, tyrosine) may increase
production of toxic Clostridia metabolites
Summary of Clostridia dopamine
abnormalities in autism
• Elevated HPHPA and similar phenolic compounds from
multiple species of Clostridia are elevated in more than
two-thirds of children with autism.
• Elevated HPHPA and cresol inhibit the conversion of
dopamine to norepinephrine, resulting in marked
increase in dopamine.
• Dopamine metabolites deplete brain glutathione and also
produce marked excess of oxygen superoxide free
radicals, 2500 molecules for each dopamine metabolite
• Brain and sympathetic nervous systems normally using
norepinephrine switch to dopamine, drastically altering
brain and sympathetic nervous system function
Summary of Clostridia dopamine
abnormalities in autism
• Presence of this abnormality is determined with HPHPA,
cresol, and other phenolic compounds in the Great Plains
organic acid test
• Stool testing not useful since HPHPA producing and NonHPHPA species are not differentiated
• Glutathione depletion by excess dopamine determined by
pyroglutamic acid marker in organic acid test.
• Clostridia treated for 10-14 days with vancomycin or
flagyl,followed by several months high dose probioticsculturelle or VSL-3
• Repeat testing every 3 months to prevent recurrence