os efeitos do metabolismo de bactérias clostridium sobre a ... 2015 clostridia... · os efeitos do...
TRANSCRIPT
Os efeitos do metabolismo de bactérias Clostridium sobre a função
gastrointestinal e saúde mental e equilíbrio neurotransmissor
William Shaw Ph.D.The Great Plains Laboratory
www.GPL4U.com
Dialogues Clin Neurosci. 2011;13:55-62.
Nature Reviews Neuroscience | AOP, published online 12 September 2012;
Nature Reviews Neuroscience AOP, published online 12 September 2012;
16050–16055 | PNAS | September 20, 2011 | vol. 108 | no. 38
• “Foi observador que doentes mentais, em geral, parecem excretar quantidades muito maiores de HPHPA do que pessoas normais."
• A maioria dos pacientes com atraso mental excreta quantidades muito baixas de HPHPA.
Journal of Biological Chemistry 225:269-278,1957William Shaw PhD
Nutritional Neuroscience 2010 Vol 13 No 3: 1-10
Cresol urinária elevado é associado com maior gravidade clínicaindependentemente do sexo, e história de regressão comportamental
Estrutura de Fenóis produzidos porespécie de clorstrídia
C.Difficile somenteMultiplas espécies de Clostridia
Shaw, W Increased Urinary Excretion of Analogs of Krebs Cycle Metabolites and Arabinose in Two Brothers with Autistic Features. Clin Chem 41:1094-1104, 1995
HPHPA
HPHPAHPHPA
Psicose infantil durante internamento(simulado da memória)
Endoscopia do colon de um paciente com proliferação severa de clostridia - colite
pseudomembranosa
colite pseudomembranosa
colite pseudomembranosa
Clostridia por elétron microscopia
Clostridia por elétron microscopia
J Child Neurol 2000 Jul;15(7):429-35Short-term benefit from oral vancomycin treatment of regressive-onset autism.Sandler RH, Finegold SM, et al
•11 crianças com autismo regressivo foram recrutados para um intervenção usando vancomicina.
•Um psicólogo clínico cego ao estatus do tratamento, notoumelhora em 8 de 10 pacientes estudados.
•“…estes resultados indicaram que uma possível conexão do flora intestinal- cérebro requer mais investigação.”
Espécies de Clostrídia que sabemos que produzemprecursores de HPHPA
Shaw, Nutritional Neuroscience 2010 Vol 13 No 3: 1-10
• C. sporogenes• C. botulinum-
envenenamentoalimentar
• C. caloritolerans
• C. mangenoti• C. ghoni• C. bifermentans
Estatus dos cerca de 80 outras espécies de Clostridia no trato GI desconhecido
Espécies de Clostrídia que sabemos que não produzem precursoresde HPHPA
Nutritional Neuroscience 2010 Vol 13 No 3: 1-10
• C. tetani• C. sticklandii• C. lituseburense• C. subterminale• C. putifaciens• C. propionicum• C. malenomenatum
• C. difficile• C. histolyticum• C. aminovalericum• C. sporospheroides• C. limosum• C. lentoputrescens• C. tetanomorphum
Estatus dos cerca de 80 outras espécies de Clostridia no trato GI desconhecido
Síntese da Catecolamina
Catabolismo da Dopamina
(ORGANIC ACID TEST)
Acta psychiatr belg 80:249-265,1980
Main dopaminemetabolite HVAmmol/mol creatinine
p<0.01
Normal
Autism
Por que a dopamina está alta?
Goodhart PJ, DeWolf WE Jr, Kruse LI. Mechanism-based inactivation of dopamine beta-hydroxylase by p-cresol and related alkylphenols. Biochemistry. 1983 Jun 21; 22(13):3091-6.
• Uma ampla gama de fenóis, incluindo p-cresol, um metabolito principal da Clostridia, são potentes inibidores da dopamina-beta-hidroxilase. Que converte dopamina a noradrenalina
Ácido Fumárico
Efeito crítico de bactérias intestinais em neurotransmissores cerebrais
Teste de Ácidos Orgânicos
Teste de ÁcidosOrgânicos
Teste de ÁcidosOrgânicos
William Shaw PhD
Linan Chen, et al(2008) Unregulated cytosolic dopamine causes neurodegeneration associated with oxidative stress in mice. J. Neurosci. 28, 425–433
• A dopamina é uma molécula muito reactiva em comparação com outros neurotransmissores, e a degradação da dopamina naturalmente produz espécies oxidativas.
• Mais de 90% da dopamina em neurónios dopaminérgicos é armazenado em vesículas terminais abundantes e está protegido contra a degradação.
• No entanto, uma pequena fração citosólica é de dopamina, e que é a principal fonte do metabolismo da dopamina e toxicidade presumido.
Linan Chen, et al(2008) Unregulated cytosolic dopamine causes neurodegeneration associated with oxidative stress in mice. J. Neurosci. 28, 425–433
• Cytosolic dopamine undergoes degradation to HVA as well as extremely toxic dopamine metabolites and oxidative species which deplete brain glutathione
• These toxic metabolites caused by excess dopamine may cause severe neurodegeneration of neural pathways that utilize dopamine as a neurotransmitter.
Toxicity of excess dopamine
Homovanillic acid (HVA)Dihydroxyphenylacetic(DOPAC)
Cycled thousands of times
4-S-glutathionyl-5, 6-dihydroxyindoline
Aminochrome
Toxicity of high dopamine metabolite (Aminochrome)Mitochondria
Consequences of excess dopamine
• Overstimulation of dopamine tracts• Substitution of dopamine into norepinephrine
tracts and sympathetic nervous system• Damage to neurons producing excess
dopamine due to oxidative damage of abnormal dopamine metabolites and mitochondrial damage
• Depletion of glutathione in brain making it susceptible to other toxic chemicals
Dopamine
Areas of brain producing dopamine
Effect of HPHPA on neurotransmitters-severe autism-case 1
0
10
20
30
40
50
60
70
80
0
1000
2000
3000
4000
5000
6000
7000
8000
9000
6/1/2008 12/18/2008 7/6/2009 1/22/2010 8/10/2010 2/26/2011 9/14/2011 4/1/2012 10/18/2012 5/6/2013
HPHPA
HVA
VMA
VMA (norepinephrine)
HVA (dopamine)
HPHPAClostridia
HPHP
A m
mol
/mol
crea
tinin
e
HVA,
VMA
mm
ol/m
ol cr
eatin
ine
Time (calendar date)
Effect of HPHPA on neurotransmitters-severe autism-case 2
HPHP
A m
mol
/mol
crea
tinin
e
HVA,
VMA
mm
ol/m
ol cr
eatin
ine
Time (calendar date)
0
5
10
15
20
25
30
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
11/18/2010 2/26/2011 6/6/2011 9/14/2011 12/23/2011 4/1/2012 7/10/2012 10/18/2012
HPHPA
HVA
VMA
HVA (dopamine)
VMA (norepinephrine)
HPHPAClostridia
William Shaw PhD
Norepinephrine
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)
Properties of Clostridia bacteria• Strict anaerobe-dies when exposed to oxygen• Cause tetanus, diarrhea, and botulism (food poisoning)• Forms spores that are highly resistant to heat and
antibiotics• About 100 species of Clostridia in GI tract• Controlled by vancomycin, metronidazole, and
Lactobacillus acidophilus GG
Spores
Spore formation-most difficult challenge to prevent recurrence
• Recorded as many as 20 recurrences after use of metronidazole/vancomycin
• Spores not killed by common disinfectants like alcohol hand wipes-may actually help spores spread
• Only bleach kills spores-MMS-mucosa damage• Carriers without symptoms may spread spores
William Shaw PhD
Effect of metronidazole on urine HPHPA levels in autismWilliam Shaw Nutritional Neuroscience 2010 Vol 13 No 3: 1-10
-50
0
50
100
150
200
250
300
350
400
450
500
0 20 40 60 80 100
Days after start of treatment
Urin
e HP
HPA
mm
ol/m
ol c
reat
inin
eStart metronidazole
Stopmetronidazole
Alternative treatments for Clostridium difficile disease: what really works? Lynne V. McFarland Journal of Medical Microbiology (2005), 54, 101–111
125–500 mg every 3 days over a meanof 27 days
44 yr female with severe anorexia nervosa
44 yr female-severe anorexia nervosa
HVA/VMA= 5:1
DopamineNorepi, Epi
Mental retardation -Altered dopamine/norepinephrine ratio with
Clostridia HPHPA excess
Dopamine/ Norepi, Epi
Vegetarian schizoid patient
Early Parkinson’s disease
65 y.o. F-Depression ,irritable bowel, before treatment
Adult female
65 y.o. F-Depression and irritable bowel resolved after treatment of Clostridia
Adult female
6 yr old female-Severe aggression-odd behavior
Two months of nystatin and Lactobacillus acidophilus GG therapy in a child with autism
Candida krusei stool
Yeast tartaric urine*
Lacto-bacillus stool
Clostridia HPHPA urine*
Before
4+
993
0
3265
After
0
1
4+
174
normal range
0-1+
0-15
3+ - 4+
0-150* mmol/mol creatinine
Effect of anti-Clostridia therapy on urine excretion of HPHPA* in
young woman with acute psychosis-auditory
hallucinations patient normals During acute 7489 0-150 psychosis After treatment 673 0-150 ( depressed but no psychosis) *mmol/mol creatinine
patient normals Before treatment 1444 0-150
After treatment 13 0-150 *mmol/mol creatinine
Effect of anti-Clostridia therapy on urine excretion of HPHPA* in woman with
depression and chronic fatigue
Treatments for Clostridia bacteria• Vancomycin-oral-not intravenous-5-10 mg/Kg/day div into 3
doses-10 days• 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
Side effects of metronidazole• Metallic taste• Nausea• Antabuse effect • Rare cases of neurotoxic complications-almost
all resolved after stopping drug:• Cerebellar dysfunction, ataxia• Altered mental status• Seizures
0.00
200.00
400.00
600.00
800.00
1000.00
1200.00
1400.00
1600.00
1800.00
2000.00
N = 14
N = 30
N = 211
CONTROLMALE
INFANTS
CONTROLMALE
CHILDREN AUTISTICMALES
3-(3
-hyd
roxy
phen
yl)-3
-hyd
roxy
-pr
opio
nic
acid
(M
MO
L/M
OL
CR
EATI
NIN
E)
2 - 13 YRS 2 - 13 YRS
Values for HPHPA Clostridia metabolite in urine samples of male infants, control boys, and boys with autism. W Shaw Nutr Neuroscience 2010 Vol 13 No 3: 1-10
Controlinfants
Control boys2-13 years
Autistic Boys2-13 years
HPHPA Clostridia metabolite in urine samples in autismW Shaw Nutr Neuroscience 2010 Vol 13 No 3: 1-10
• In unselected autistic male children (n=211), the mean value was double that of the controls (n=30) while the median value was triple that of the controls.
• Similar results were found for females. • The differences between autism and control groups of
the appropriate sex were statistically significant by the t-test at P < 0.005.
• The HPHPA values in urine samples of more than two thirds of the children with autism exceeded control mean and median values.
Comparisons to other tests
• Urine profiles of neurotransmitters without valid Clostridia marker gives false picture of neurotransmitter metabolism
• Many Clostridia are beneficial bacteria so stool testing may give erroneous results
• Only GPL organic acid test containing HPHPA gives correct information of both Clostridia and neurotransmitter metabolites
Great Plains organic acid test-child with autism
Same child as previous slide-DNA stool test
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,epinephrine- 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 sulfo-transferase (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
• Pyridoxic acid –B6-cofactor for DOPA decarboxylase
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 difficile-blocks 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
Incidence of Clostridia by organic acid urine testing, n=1000 consecutive urine organic
acid samplesHPHPA
+4-Cresol
+HPHPA and
4-Cresol+
15.2% 6.8% 1.6%
Totalpositives=
23.6%
Clinical usefulness of Clostridia treatments
• Schizophrenia• Psychosis• Depression• Chronic fatigue• Tics, Tourette’s• Autism• Parkinson’s disease• Arthritis
• ADD, ADHD• Obsessive compulsive
disorder• Seizure disorders• Irritable bowel• Crohn’s disease• Ulcerative colitis
