The Biochemical Basis Autism Spectrum Disorders

The Biochemical Basis of Autistic Disorders : The Mercury, Androgen (Testosterone), & Glutathione Connection Mark R. Geier, MD, PhD, FABMG, FACE President The Genetic Centers of America Phone: (301)989-0548 Email: [email protected]

David A. Geier Vice-President The Institute of Chronic Illnesses, Inc. Copyright 2008

Mercury Exposure Background Information

Thimerosal & Vaccines: 

Thimerosal is an organic mercury compound (50% mercury be weight) that is metabolized to ethylmercury and thiosalicylate and has been present since the 1930s as a preservative in some vaccines and pharmaceutical products to prevent bacterial and fungal contamination.

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The FDA in 1999, under the recommended childhood immunization schedule, determined infants might be exposed to cumulative doses of ethylmercury that exceed some federal safety guidelines established for exposure to methylmercury, another form of organic mercury.

Thimerosal: The Early History Source: Geier DA, Sykes LK, Geier MR. A Review of Thimerosal (Merthiolate) and its Ethylmercury Breakdown Product: Specific Historical Considerations Regarding Safety & Effectivness. Journal of Toxicology & Environmental Health: Part B Critical Reviews 2007;10:575-96.

Smithburn KC, Kempf GF, Zerfas LG, Gilman LH. Meningococcic Meningitis: A Clinical Study of 144 Epidemic Cases.* Journal of the American Medical Association 1930

“TREATMENT The treatment has remained essentially the same throughout the epidemic. The routine adopted included, on admission, a skin sensitization test, rhacehicentesis and intratheceal serum intramuscular serum and (especially during the second month of the epidemic) serum intravenously. Intravenous administration of an antiseptic solution was tried and found wanting despite the in vitro activity of the agent.”

* From the Lilly Laboratories for Clinical Research, Indianapolis City Hospital The bacteriologic and serologic studies were made by H. M. Powell, A.B., Sc.D., and F. G. Jones, of the biologic department of the Lilly Research Laboratories.

Powell HM, Jamieson WA. Merthiolate as a Germicide.* American Journal of Hygiene 1931 “Toxicity in Man. Merthiolate has been injected intravenously into 22 persons in doses upt to 50 cubic centimeters of 1 per cent solution. As many as five intravenous doses, or a total of 180 cubic centimeters of 1 per cent Merthiolate, have been given to one individual (see table 7). These large doses did not produce any anaphylacotid or shock symptoms. Neither did these quantities in the repeated doses bring about any demonstrable later toxic effects. The toleration of such intravenous doses indicates a very low order of toxicity of Merthiolate for man. This information has been supplied through the kindness of Dr K. C. Smithburn of Indianapolis who has had occasion to use Merthiolate in a clinical way. Dr. Smithburn state that in these cases ‘beneficial effect of the drug was not definitely proven. It did not appear however to have any deleterious action when used in rather large doses intravenously when all the drug entered the vein.’” * From the Research Laboratories, Eli Lilly Company, Indianapolis, Indiana. We wish to express our thanks to Dr. G. H.A. Cloves; Director of Research of the Eli Lilly Company, for assistance and suggestions in the course of this investigation.

Letter from the Director of Biological Laboratories of Pitman-Moore Company to W. A. Jamieson, Director of Biological Division, Eli Lilly & Company (July 22, 1935) “We have obtained marked local reaction in about 50 percent of the dogs injected with serum containing dilutions of Merthiolate varying from 1 in 40,000 to 1 in 5,000…no connection between the lot of serum and the reaction. In other words, Merthiolate is unsatistifactory as a preservative for serum intended for use on dogs…”

Engley FB. Mercurials as Disinfectants: Evaluation of Mercurial Antimicrobic Action & Comparative Toxicity for Skin Tissue Cells. Presented May 21, 1956 at 42nd Midyear Meeting of the Chemical Specialties Manufacturers Association, Chicago, IL.

The Dose, Makes the Poison: Late 1980s to Early 2000s: •

Rh-negative mothers were routinely administered Thimerosal containing Rho(D)-immune globulins at 28 weeks gestation (10.5 to in some instances more than 40 µg mercury per dose).

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Infants may have been exposed a total of 237.5 µg mercury during the first 18-24 months of life, if all Thimerosal-containing vaccines were administered.

Early 2000s – Present: •

All pregnant women to receive flu vaccine anytime during pregnancy (25 µg mercury / dose).

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Infants to receive 3 flu vaccines during first 18-24 months of life (12.5 µg mercury / dose) = 37.5 µg mercury.

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Children from 3 years-old through 18 years-old are to receive yearly flu vaccines (25 µg mercury / dose) = 375 µg mercury.

● US Hot Spots Have Mercury Levels in Excess of US Government Regulations Source: Commission for Environmental Cooperation, 2008.

Thimerosal (Mercury) Doses** Infants Received: Source: Bigham M, Copes R. “Thiomersal in vaccines: balancing the risk of adverse effects with the risk of vaccine-preventable disease,” Drug Saf, 2005;28:89-101.

__________________________ ** Assuming an infant receiving 187.5 μg of mercury from Thimerosalcontaining vaccines during the first 6 months of life from the routine childhood vaccination schedule, in combination with environmental exposure from mercury in breast milk (164 μg of mercury).

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“Learning and developmental disabilities (LDDs) include but are not limited to deficits in learning and memory, reduced IQ, attention deficit hyperactivity disorder (ADHD), autism spectrum disorder, conduct disorders and developmental delays.”

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“There is no doubt that mercury exposure causes learning and developmental disorders”

Increased Mercury Exposure in Autistic Disorders

The Centers For Disease Control & Prevention (CDC) Vaccine Safety Datalink (VSD) Thimerosal Dose-Response Studies

Simpsonwood Meeting (7-8 June 2000) in Norcross, GA where the findings of the Vaccine Safety Datalink (VSD) analysis showing a link between Thimerosal-containing vaccines and neurodevelopmental outcomes were discussed in a closed meeting by employees from the CDC, FDA, & the vaccine manufacturers.

Dr. B renier: Page 113: “We hav e as ked you t o keep t his inf ormat ion confident ial…” Dr. Johnston: Page 198: “F orgi ve this personal comm ent, but I got cal led out a eight o’c lock emergency call and m y daught er-in-law delivered a son by C-Secti on. Our f irs t male in the line of the next generat ion, and I do not want t hat grands on to get a thimerosal containing vacc ine” Dr. W eil: Page 207: “The num ber of dos e related relat ions hi ps are li near and stat isti cally signif ic ant . You can play with thi s all you want. They are linear. They are stat is tically s ignifi cant . The pos it ive relat ions hi ps are thos e that one might expec t f rom the Faeroe I slands studi es . They are als o relat ed t o t hos e dat a we do hav e on experim ental animal dat a and simi lar to the neurodev el opment al t ox data on other substances , so that I think you can’t ac cept that this is out of the ordinary. It is n’t out of the ordinary. ” Dr. B rent: Page 229: “…w e are in a bad pos it ion from t he standpoi nt of def ending l aw sui ts if they were i ni tiat ed and I am conc erned. ”

Dr. Clem ents: Page 247: “I am really conc erned that we have tak en of f like a boat going dow n one arm of the mangrove sw am p at high speed, when in f ac t there was not enough disc uss ion really early on about whic h way t he boat should go at all. And I reall y do want to risk of fending ev eryone i n t he room by say ing that perhaps thi s study should not have been done at all, becaus e the out com e of it could have t o some ex tent, been predicted, and w e have all reached t his point now where we are lef t hanging…” “… But nonet heles s, we know f rom many experienc es in history t hat the pure scienti st has done res earch becaus e of pure scienc e. But that pure sc ience has result ed in splitt ing the at om or some other proces s which is com pletely beyond t he pow er of the s cient is ts who did the res earch t o cont rol i t. And what we hav e here is peopl e who have, for ev ery best reas on in the worl d, purs ued a di rec ti on of researc h. But t here i s now the point at which t he researc h res ult s have t o be handled, and ev en if t his c om mit tee decides that there is no as sociat ion and that inf ormat ion gets out , the work t hat has been done and t hrough t he freedom of inform ation that wil l be tak en by ot hers and will be us ed i n way s bey ond the control of t his group. An I am very conc erned about that as I suspec t i t is already t oo l ate to do anything regardles s of any profess ional body and what t hey say. My mandat e as I sit here i n t his group is to make sure at t he end of the day t hat 100,000, 000 are im munized wit h DT P, Hepatit is B and i f poss ible Hib, this year, next year and f or many years t o come, and

A Prospective Assessment of the Association between ThimerosalContaining Rho(D)-Immune Globulin and Autistic Disorders By Geier DA, Geier MR Journal of Maternal, Fetal, & Neonatal Medicine 2007;20:385-90

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Increased Mercury Body-Burden in Autistic Disorders: A Clinical Perspective

Conclusions: ** There was a significant increase in the brain concentration of the Hg / Se ratio in autistics vs controls. ** There was a significant increased in brain oxidative stress markers in autistics vs controls. ** There was a significant correlation between brain mercury concentrations and oxidative stress markers in autistics vs controls.

Low Glutathione in Autistic Disorders

Methionine Cycle & Transsulfuration Pathway Source: James SJ, et al**. Metabolic biomarkers of increased oxidative stress and impaired methylation capacity in children with autism. Am J Clin Nutr. 2004 Dec;80(6):1611-7.

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** Mercury Excretion is Directly Related to Glutathione Secretion

Source: Clarkson TW, Nordberg GF, Sager PR. Reproductive and developmental toxicity of metals. Scan J Work Environ Health 1985;11:145-54.

Biochemical Markers in Autistics: Sourece: James SJ, et al. Metabolic biomarkers of increased oxidative stress and impaired methylation capacity in children with autism. Am J Clin Nutr. 2004 Dec;80(6):1611-7.

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Visual Evidence of Thimerosal Induced Human Neuron Damage

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↑ Human Neuroblastoma Cells 24 Hrs Incubation No Thimerosal

Human Neuroblastoma Cells 24 Hrs Incubation 100 nM Thimerosal [20 ppb Mercury]

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↑ Human Fetal Cells 24 Hrs Incubation No Thimerosal

Human Fetal Cells 24 Hrs Incubation 10 nM Thimerosal [2 ppb Mercury]

Mercury & Testosterone Toxicity

** Observed that female hormones afforded total protection against Thimerosal toxicity. ** Observed testosterone at 1.0 µM levels that by itself did not significantly increase neuron death, within 3 hours when added with 50 nM Thimerosal caused 100% neuron death [50 nM Thimerosal at this time point did not significantly cause any cell death].

Geier MR, Geier DA. The potential importance of steroids in the treatment of autistic spectrum disorders and other disorders involving mercury toxicity. Med Hypotheses 2005;64:946-54.

Mannin g J T, Baro n-Cohen S, Wheel wri ght S, Sand ers G. T he 2nd t o 4t h digit rat io and a ut ism. D ev Med Chi ld Neurol 200 1;43:16 0-4 . The aut hor s ex am ined 7 2 chi ldre n wi th aut ism , inc lu ding 2 3 chi ldren wi th Asp erg er sy nd ro me, 3 4 si bli ng s, 88 fat hers, 8 8 mot hers, and sex and a ge-mat ched cont ro ls. The aut ho rs dem onst rat ed that the m ore se verel y a ffec ted t he chi ld ren were t he hi gher the lev els of prenat al test ost erone. Low er Testo ste ro ne/ Hig he r E stro gen

Hig her Tes tostero ne / Low er Es troge n AS = As perg er Syn dro me

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Hydroxysteroid Sulfotransferase

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Elevated Testosterone (Androgens) in Children with Neurodevelopmental Disorders: Potential Insights into Levels & Potential Adverse Effects

Developmental Medicine & Child Neurology 1999;41:392–395

Discussion

Studies on precocious puberty have primarily focused on children with typical patterns of growth and cognitive development. This study reviewed diagnostic data from the records of 15,719 patients with neurodevelopmental disabilities for diagnoses associated with premature sexual development/precocious puberty. Thirty-two individuals with premature sexual development were identified…

The US Department of Health and Human Services and the National Institute of Child Health and Development (NICHD) of the National Institutes of Health (NIH) estimate the incidence of precocious puberty in the general population to be approximately one in 10,000 children (US Department of Health and Human Services 1997). The incidence of precocious puberty has been estimated to be higher in children with neurodevelopmental disabilities than in children without neurodevelopmental disabilities. Our retrospective review of this population with neurodevelopmental disabilities suggested that a child with a neurodevelopmental disability was at least 20 times more likely to experience early pubertal changes.

Am J Psychiatry 1997;154:1626-7

• In 4 of 12 prepubertal autistic children (6–10 years old) in our inpatient child psychiatry department, we have observed precocious secondary sexual characteristics (growth of pubic hair, increase of testis volume) that suggest high androgenic activity in infantile autism. • To test our hypothesis of a hyperandrogeny and autism association, we measured plasma testosterone and adrenal androgen in nine drug-free inpatients with DSM-IV autism and 62 normal subjects of same age, sex, weight (within 2 kg), and stage of puberty. • Results showed that three of the nine autistic subjects had an abnormally high plasma testosterone concentration (over two standard deviations above the mean for the comparison subjects), with values above that of the highest in the comparison subjects.

Other Effects of Elevated Testosterone (Androgens) in ASDs

• Testosterone levels are positively correlated with a number of autistic traits and inversely correlated with social development and empathy. • A medical questionnaire was completed by n=54 women with ASDs, n=74 mothers of children with ASDs, and n=183 mothers of typically developing children. • Compared to controls, significantly more women with ASDs reported (a) hirsutism, (b) bisexuality or asexuality, (c) irregular menstrual cycle, (d) dysmenorrhea, (e) polycystic ovary syndrome, (f) severe acne, (g) epilepsy, (h) tomboyism, and (i) family history of ovarian, uterine, and prostate cancers, tumors, or growths. • Compared to controls, significantly more mothers of ASD children reported (a) severe acne, (b) breast and uterine cancers, tumors, or growths, and (c) family history of ovarian and uterine cancers, tumors, or growths. • These results suggest current hormone abnormalities in women with ASC and their mothers.

A summary of the interaction between the transsulfuration and androgen pathways in autistic spectrum disorders

PAPS = 3’-phosphoadenosine 5’-phophosulfate BHMT = Betaine Homocysteine Methyltransferase MS = Methionine Synthase SAM = S-adenosylmethionine MTase = Methyltransferase SAH = S-adenosylhomocysteine

CBS = Cystathionine β-Synthase THF = Tetrohydrofolate 5-MTHF = 5-Methyltetrahydrofolate 5, 10-MTHF = 5, 10-Methyltetrahydrofolate SAHH = SAH Hydrolase DHEA-S = Dehydroepiandrosterone-sulfate DHEA = Dehydroepiandrosterone

Treatment Overview: The Protocol

• Furthermore, in our own clinical experience we have observed that leuprolide acetate (LUPRON®) administration to nearly 200 patients diagnosed with ASDs significantly lowered androgen levels and has resulted in very significant overall clinical improvements in socialization, sensory/cognitive awareness, and health/physical/behavior skills, with few non-responders and minimal adverse clinical effects to the therapy. • The following are some specific areas of significant clinical ameliorations in frequent symptoms that occur in patients diagnosed with ASDs observed: • hyperactivity/impulsivity • stereotypy • aggression • self-injury • abnormal sexual behaviors • irritability behaviors