Michael Aschner & Sandra Ceccatelli: Sins of Omission Regarding Thimerosal

Teresa Binstock
Researcher in Developmental & Behavioral Neuroanatomy
December 04, 2009


Introduction: A new research paper by Aschner & Ceccatelli is supposedly peer-reviewed but has sufficient omissions so as perhaps to merit consideration as scientific fraud, perhaps to be considered as injurious to children who have increased susceptibility -- whether genetic and/or acquired -- for adverse effects from mercury, methylmercury, and ethylmercury.

Michael Aschner, Ph.D., is a professor of neuroscience, pediatrics, and pharmacology at Vanderbilt University (1) and has published more than 90 research articles about mercury (2). One of his more recent papers (3) is coauthored with Sandra Ceccatelli, M.D., Ph.D. (4), who has published at least 26 papers about mercury (5).

Despite these extensive backgrounds, Aschner & Ceccatelli 2009 (3) omit studies which have found associations among thimerosal, autism, and special education services (cites below).  Furthermore, Aschner & Ceccatelli err in stating (as if factual) that thimerosal in vaccines "was removed from the US market in 2001." (3). Despite the erroneous statement by Aschner & Ceccatelli, thimerosal has been included in some U.S. vaccines in each of the years from 2002 to 2009 (6).

Indictment: Here is a list of errors and significant omissions in Aschner & Ceccatelli 2009 (3).

1. Contrary to the supposedly peer-reviewed statement by Aschner & Ceccatelli, thimerosal has been available in U.S. vaccines each and every year since 2001 (6). In recent years and currently, most influenza vaccines and most H1N1 vaccines contain thimerosal (6), which is associated with autism and other developmental disabilities (cites below).

2. Aschner & Ceccatelli dwell upon Pichichero et al's 2002 findings regarding levels of ethylmercury injected as thimerosal into healthy infants (7) but fail to cite and discuss Waly et al 2004, who found that intra-body thimerosal levels lower than those described by Pichichero et al were sufficient to impair methionine synthase, an enzyme important in neurodevelopment (7). Ironically, contrary to Aschner's & Ceccatelli's erroneous declaration that thimerosal was removed in 2001, Pichichero et al 2002 used thimerosal-containing vaccines, "diptheria-tetanus-acellular pertussis vaccine, hepatitis B vaccine, and in some children Haemophilus influenzae type b vaccine" (8), a research methodology consistent with thimerosal remaining in various U.S. vaccines (6).

3. Aschner & Ceccatelli offer a summary entitled "Use of Thimerosal in Vaccines" (3; p2) but omit peer-reviewed summaries which reported thimerosal's lack of efficacy. For instance:  "...mercurials are ineffective in vivo and may be more toxic for tissue cells than bacterial cells, as shown in mice, tissue culture and embryonic eggs and with leucocytes " (10, ANYAS; listed in 9) and " Thimerosal is a weak antibacterial agent that is rapidly broken down to products, including ethylmercury residues, which are neurotoxic. Its role as a preservative in vaccines has been questioned, and the pharmaceutical industry itself considers its use historical." (11, Lancet; listed in 9).
    Furthermore, Aschner & Ceccatelli cite Powell & Jamieson 1931 but fail to cite numerous studies whose findings call attention to thimerosal's adverse effects and to thimerosal's quite imperfect efficacy as bactericidal or bacteriostatic. Indeed, Aschner & Ceccatelli failed to find or deliberately ignored a thorough thimerosal-review published in the Journal of Toxicology and Environmental Health (12). The review by Geier et al is in the PubMed database, is free online (12), and summarizes findings which contradict Aschner & Ceccatelli's glib oversimplification regarding thimerosal.

4. Aschner & Ceccatelli omit CDC findings by Verstraeten et al 1999, the original CDC-sponsored study which found associations between thimerosal injections and a range of developmental disabilities including but not limited to autism, tics, and speech disorders (13). After a series of deliberate dilutions of their own data, the CDC group published a sanitized rendition as Verstraeten et al 2003 (14).  Obtained by Freedom of Information Act request (13), the CDC's 1999 findings stand in stark contrast to the diluted-data findings presented in Pediatrics (14).

5. Aschner & Ceccatelli omit findings by Goodman & Gallagher, Stony Brook medical school researchers who found that, among boys, early life thimerosal injections are associated with an increased likelihood of autism-spectrum disorder and with an increased need for special education services (15-16).

6. Aschner & Ceccatelli omit findings by James SJ & colleagues, who have reported associations among thimerosal, autism, and metabolic pathways related to glutathione (eg, 17-18) and omit other studies documenting the bidirectional genetics of thimerosal detoxification (eg, 19-20).

7. Aschner & Ceccatelli omit findings about thimerosal's adverse effects upon mitochondria (eg, 17, 21-24), despite the autism significance of mitochondrial disorders (eg, 25-27) and mitochondrial dysfunction not necessarily genetic in origin (reviewed in 28).

8. Aschner & Ceccatelli omit findings about porphyrins and mercury in autism (eg, 29-30).


Discussion: The erroneous and misleading article by Aschner & Ceccatelli (3) prompts questions.  How or why did such experienced professors, the article's reviewers, and the journal's editor (31) err in regard to thimerosal's continued presence in some vaccines injected into children in the United States?  How and why did Aschner & Ceccatelli choose to omit studies with findings which contradict the rationale the authors chose to impose?

Ceccatelli & Aschner have considerable experience in writing about mercury, thus their seemingly well documented rationale about ethylmercury creates a convincing but misleading illusion. Clearly, the content and omissions in Aschner & Ceccatelli suggest that the journal Neurotoxicity Research needs publish a correction of factual errors in Ceccatelli & Aschner and ought publish a paper which remedies the omissions and rebuts the implications cleverly but inaccurately presented by Aschner & Ceccatelli.

Physicians, researchers, and others are encouraged to read Aschner's and Ceccatelli's summary entitled "Use of Thimerosal in Vaccines" (p2 in 3) and to compare their overview with  thimerosal findings summarized by Geier et al (12).

Given these flaws in Aschner & Ceccatelli 2009, two caveats are merited. Firstly, the abstract states, "In this review, we have discussed the hypothesis that exposure to thimerosal during childhood may be a primary cause of autism. The conclusion is that there are no reliable data indicating that administration of vaccines containing thimerosal is a primary cause of autism." [sic, eg, 15-17] Secondly, in their abstract the authors qualify the gist of their skillfully misleading presentation by admitting, "...one cannot rule out the possibility that the individual gene profile and/or gene–environment interactions may play a role in modulating the response to acquired risk by modifying the individual susceptibility."  Despite the authors' declaration of this "possibility", Aschner's and Ceccatelli's deliberate omissions and their masterfully crafted illusion of science suggest the authors have made the moral equivalent of a Faustian bargain.




References
:

1. Michael Aschner, Ph.D.
http://kc.vanderbilt.edu/site/people/8993/aschner-michael.aspx

2. Pubmed, Nov 20, 2009
Aschner M AND (mercury OR methylmercury OR ethylmercury OR thimerosal)

3. Are Neuropathological Conditions Relevant to Ethylmercury Exposure?
Michael Aschner and Sandra Ceccatelli
http://www.springerlink.com/content/3398g44388158630/fulltext.pdf

4. Sandra Ceccatelli, M.D., Ph.D.
http://ki.se/ki/jsp/polopoly.jsp?d=21984&a=54602&l=en

5. PubMed, Nov 20, 2009
Ceccatelli S AND (mercury OR methylmercury OR ethylmercury OR thimerosal)

6. Thimerosal content in some US vaccines (yearly summaries, 2002 through 2009)
Institute for Vaccine Safety (IVS); Johns Hopkins Bloomberg School of Public Health
http://www.vaccinesafety.edu/
IVS data summaries dated: 02-21-2002; 12-17-2002; 6/20/2003; 04/22/2004; October 2004; February 2005; Sept 13; 2005; Oct 16, 2006; 7/11/07; 5/15/08; 8/6/08; 10/19/09/ 11/19/09.

7. Mercury concentrations and metabolism in infants receiving vaccines containing thiomersal: a descriptive study
Pichichero ME, Cernichiari E, Lopreiato J, Treanor J.
Lancet. 2002 Nov 30;360(9347):1737-41.

8. Activation of methionine synthase by insulin-like growth factor-1 and dopamine: a target for neurodevelopmental toxins and thimerosal
Waly M et al.
Mol Psychiatry. 2004 Apr;9(4):358-70.

9. Thimerosal is an Ineffective Vaccine Preservative
http://www.autismboulder.org/pdf/IneffectivePreservative.pdf

10. Evaluation of mercurial compounds as antiseptics
Engley FB
Annals of New York Academy of Sciences. 53:197-2-6, 1950.

11. The case against thimerosal use
Seal D et al.
Lancet, 1991;338:315-316.

12. A review of Thimerosal (Merthiolate) and its ethylmercury breakdown product: specific historical considerations regarding safety and effectiveness
Geier DA, Sykes LK, Geier MR.
J Toxicol Environ Health B Crit Rev. 2007 Dec;10(8):575-96.
{free online}
http://www.informaworld.com/smpp/content~db=all?content=10.1080/10937400701389875

Thimerosal (Merthiolate) is an ethylmercury-containing pharmaceutical compound that is 49.55% mercury and that was developed in 1927. Thimerosal has been marketed as an antimicrobial agent in a range of products, including topical antiseptic solutions and antiseptic ointments for treating cuts, nasal sprays, eye solutions, vaginal spermicides, diaper rash treatments, and perhaps most importantly as a preservative in vaccines and other injectable biological products, including Rho(D)-immune globulin preparations, despite evidence, dating to the early 1930s, indicating Thimerosal to be potentially hazardous to humans and ineffective as an antimicrobial agent. Despite this, Thimerosal was not scrutinized as part of U.S. pharmaceutical products until the 1980s, when the U.S. Food and Drug Administration finally recognized its demonstrated ineffectiveness and toxicity in topical pharmaceutical products, and began to eliminate it from these. Ironically, while Thimerosal was being eliminated from topicals, it was becoming more and more ubiquitous in the recommended immunization schedule for infants and pregnant women. Furthermore, Thimerosal continues to be administered, as part of mandated immunizations and other pharmaceutical products, in the United States and globally. The ubiquitous and largely unchecked place of Thimerosal in pharmaceuticals, therefore, represents a medical crisis.

13. [analysis] Generation Zero
Blaxill M, Safeminds 2004
http://www.safeminds.org/research/library/GenerationZeroPowerPoint.pdf

14. Safety of thimerosal-containing vaccines: a two-phased study of computerized health maintenance organization databases
Verstraeten T, Davis RL, DeStefano F, Lieu TA, Rhodes PH, Black SB, Shinefield H, Chen RT; Vaccine Safety Datalink Team.
Epidemic Intelligence Service Program, Epidemiology Program Office
Centers for Disease Control and Prevention
Pediatrics. 2003 Nov;112(5):1039-48.
http://pediatrics.aappublications.org/cgi/content/full/112/5/1039

15. Hepatitis B vaccination of male neonates and autism
[conference abstract as published]
CM Gallagher, MS Goodman, Graduate Program in Public
Health, Stony Brook University Medical Center, Stony Brook, NY
Annals of Epidemiology, p659
Vol. 19, No. 9 Abstracts (ACE) September 2009: 651–680

PURPOSE: Universal newborn immunization with hepatitis B vaccine was recommended in 1991; however, safety findings are mixed. The Vaccine Safety Datalink Workgroup reported no association between hepatitis B vaccination at birth and febrile episodes or neurological adverse events. Other studies found positive associations between hepatitis B vaccination and ear infection, pharyngitis, and chronic arthritis; as well as receipt of early intervention/special education services (EIS); in probability samples of
U.S. children. Children with autistic spectrum disorder (ASD) comprise a growing caseload for EIS. We evaluated the association between hepatitis B vaccination of male neonates and parental report of ASD.
METHODS: This cross-sectional study used U.S. probability samples obtained from National Health Interview Survey 1997–2002 datasets. Logistic regression modeling was used to estimate the effect of neonatal hepatitis B vaccination on ASDrisk amongboys age 3–17 years with shot records, adjusted for race, maternal education, and two-parent household.
RESULTS:Boyswho received the hepatitis B vaccine during the first month of life had 2.94 greater odds for ASD (nZ31 of 7,486; OR Z 2.94; p Z 0.03; 95% CI Z 1.10, 7.90) compared to later- or unvaccinated boys. Non-Hispanicwhite boys were 61% less likely to have ASD(ORZ0.39; pZ0.04; 95% CIZ0.16, 0.94) relative to non-white boys.
CONCLUSION: Findings suggest that U.S. male neonates vaccinated with hepatitis B vaccine had a 3-fold greater risk of ASD; risk was greatest for non-white boys.

16. Hepatitis B triple series vaccine and developmental disability in US children aged 1-9 years
 Gallagher C, Goodman M. Toxicol Environ Chem 2008 90(5):997-1008.
{free online}
http://fourteenstudies.org/pdf/hep_b.pdf

This study investigated the association between vaccination with the Hepatitis B triple series vaccine prior to 2000 and developmental disability in children aged 1-9 years (n = 1824), proxied by parental report that their child receives early intervention or special education services (EIS). National Health and Nutrition Examination Survey 1999-2000 data were analyzed and adjusted for survey design by Taylor Linearization using SAS version 9.1 software, with SAS callable SUDAAN version 9.0.1. The odds of receiving EIS were approximately nine times as great for vaccinated boys (n = 46) as for unvaccinated boys (n = 7), after adjustment for confounders. This study found statistically significant evidence to suggest that boys in United States who were vaccinated with the triple series Hepatitis B vaccine, during the time period in which vaccines were manufactured with thimerosal, were more susceptible to developmental disability than were unvaccinated boys.

17. Cellular and mitochondrial glutathione redox imbalance in lymphoblastoid cells derived from children with autism
James SJ et al.
FASEB J. 2009 Aug;23(8):2374-83.

Research into the metabolic phenotype of autism has been relatively unexplored despite the fact that metabolic abnormalities have been implicated in the pathophysiology of several other neurobehavioral disorders. Plasma biomarkers of oxidative stress have been reported in autistic children; however, intracellular redox status has not yet been evaluated. Lymphoblastoid cells (LCLs) derived from autistic children and unaffected controls were used to assess relative concentrations of reduced glutathione (GSH) and oxidized disulfide glutathione (GSSG) in cell extracts and isolated mitochondria as a measure of intracellular redox capacity. The results indicated that the GSH/GSSG redox ratio was decreased and percentage oxidized glutathione increased in both cytosol and mitochondria in the autism LCLs. Exposure to oxidative stress via the sulfhydryl reagent thimerosal resulted in a greater decrease in the GSH/GSSG ratio and increase in free radical generation in autism compared to control cells. Acute exposure to physiological levels of nitric oxide decreased mitochondrial membrane potential to a greater extent in the autism LCLs, although GSH/GSSG and ATP concentrations were similarly decreased in both cell lines. These results suggest that the autism LCLs exhibit a reduced glutathione reserve capacity in both cytosol and mitochondria that may compromise antioxidant defense and detoxification capacity under prooxidant conditions.

18. Metabolic endophenotype and related genotypes are associated with oxidative stress in children with autism
James SJ et al.
Am J Med Genet B Neuropsychiatr Genet. 2006 Dec 5;141B(8):947-56.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2610366/?tool=pubmed

Autism is a behaviorally defined neurodevelopmental disorder usually diagnosed in early childhood that is characterized by impairment in reciprocal communication and speech, repetitive behaviors, and social withdrawal. Although both genetic and environmental factors are thought to be involved, none have been reproducibly identified. The metabolic phenotype of an individual reflects the influence of endogenous and exogenous factors on genotype. As such, it provides a window through which the interactive impact of genes and environment may be viewed and relevant susceptibility factors identified. Although abnormal methionine metabolism has been associated with other neurologic disorders, these pathways and related polymorphisms have not been evaluated in autistic children. Plasma levels of metabolites in methionine transmethylation and transsulfuration pathways were measured in 80 autistic and 73 control children. In addition, common polymorphic variants known to modulate these metabolic pathways were evaluated in 360 autistic children and 205 controls. The metabolic results indicated that plasma methionine and the ratio of S-adenosylmethionine (SAM) to S-adenosylhomocysteine (SAH), an indicator of methylation capacity, were significantly decreased in the autistic children relative to age-matched controls. In addition, plasma levels of cysteine, glutathione, and the ratio of reduced to oxidized glutathione, an indication of antioxidant capacity and redox homeostasis, were significantly decreased. Differences in allele frequency and/or significant gene-gene interactions were found for relevant genes encoding the reduced folate carrier (RFC 80G > A), transcobalamin II (TCN2 776G > C), catechol-O-methyltransferase (COMT 472G > A), methylenetetrahydrofolate reductase (MTHFR 677C > T and 1298A > C), and glutathione-S-transferase (GST M1). We propose that an increased vulnerability to oxidative stress (endogenous or environmental) may contribute to the development and clinical manifestations of autism.

19. Homozygous gene deletions of the glutathione S-transferases M1 and T1 are associated with thimerosal sensitization
Westphal GA et al.
Int Arch Occup Environ Health. 2000 Aug;73(6):384-8.

OBJECTIVE: Thimerosal is an important preservative in vaccines and ophthalmologic preparations. The substance is known to be a type IV sensitizing agent. High sensitization rates were observed in contact-allergic patients and in health care workers who had been exposed to thimerosal-preserved vaccines. There is evidence for the involvement of the glutathione system in the metabolism of thimerosal or its decomposition products (organomercury alkyl compounds). Thus detoxification by polymorphically expressed glutathione S-transferases such as GSTT1 and GSTM1 might have a protective effect against sensitization by these substances. METHODS: To address this question, a case control study was conducted, including 91 Central European individuals with a positive patch-test reaction to thimerosal. This population was compared with 169 healthy controls and additionally with 114 individuals affected by an allergy against para-substituted aryl compounds. The latter population was included in order to test whether possible associations were due to substance-specific effects, or were a general feature connected with type IV immunological diseases. Homozygous deletions of GSTT1 and GSTM1 were determined by polymerase chain reaction. RESULTS: Glutathione S-transferase M1 deficiency was significantly more frequent among patients sensitized to thimerosal (65.9%, P = 0.013) compared with the healthy control group (49.1%) and the "para-compound" group (48%, P = 0.034). Glutathione S-transferase T1 deficiency in the thimerosal/mercury group (19.8%) was barely elevated versus healthy controls (16.0%) and the "para-compound" group (14.0%). The combined deletion (GSTT1-/GSTM1-) was markedly more frequent among thimerosal-sensitized patients than in healthy controls (17.6% vs. 6.5%, P = 0.0093) and in the "para-compound" group (17.6% vs. 6.1%, P =0.014), revealing a synergistic effect of these enzyme deficiencies (healthy controls vs. thimerosal GSTM1 negative individuals, OR = 2.0 [CI = 1.2-3.4], GSTT1-, OR = 1.2 [CI = 0.70-2.1], GSTM1/T1-, OR = 3.1 [CI = 1.4-6.5]). CONCLUSIONS: Since the glutathione-dependent system was repeatedly shown to be involved in the metabolism of thimerosal decomposition products, the observed association may be of functional relevance.

20. Inhibition of the human erythrocytic glutathione-S-transferase T1 (GST T1) by thimerosal
Müller M, Westphal G, Vesper A, Bünger J, Hallier E.
Int J Hyg Environ Health. 2001 Jul;203(5-6):479-81.

We have investigated the interaction of thimerosal, a widely used antiseptic and preservative, with the human erythrocytic GST T1 (glutathione-S-transferase T1). This detoxifying enzyme is expressed in the erythrocytes of solely the human species and it displays a genetic polymorphism. Due to this polymorphism about 25% of the individuals of the caucasian population lack this activity ("non-conjugators"), while 75% show it ("conjugators") (Hallier, E., et al., 1993). Using our newly developed HPLC-fluorescence detection assay (Müller, M., et al., 2001) we have profiled the kinetics of enzyme inhibition in erythrocyte lysates of two individuals previously identified as "normal conjugator" (medium enzyme activity) and "super-conjugator" (very high activity). For the normal conjugator we have determined a 2.77 mM thimerosal concentration to inhibit 50% of the GST T1 activity. In the case of the super-conjugator a 2.3 mM thimerosal concentration causes a 50% inhibition of the enzyme activity. For both phenotypes a 14.8 mM thimerosal concentration results in residual enzyme activities equal to those typically detected in non-conjugator lysates. Thus, sufficiently high doses of thimerosal may be able to change the phenotypic status of an individual--at least in vitro--by inhibition of the GST T1 enzyme.

21. Biochemical and molecular basis of thimerosal-induced apoptosis in T cells: a major role of mitochondrial pathway
Makani S et al.
Division of Basic and Clinical Immunology, University of California
Genes Immun. 2002 Aug;3(5):270-8.
http://www.nature.com/gene/journal/v3/n5/abs/6363854a.html

The major source of thimerosal (ethyl mercury thiosalicylate) exposure is childhood vaccines. It is believed that the children are exposed to significant accumulative dosage of thimerosal during the first 2 years of life via immunization. Because of health-related concerns for exposure to mercury, we examined the effects of thimerosal on the biochemical and molecular steps of mitochondrial pathway of apoptosis in Jurkat T cells. Thimerosal and not thiosalcylic acid (non-mercury component of thimerosal), in a concentration-dependent manner, induced apoptosis in T cells as determined by TUNEL and propidium iodide assays, suggesting a role of mercury in T cell apoptosis. Apoptosis was associated with depolarization of mitochondrial membrane, release of cytochrome c and apoptosis inducing factor (AIF) from the mitochondria, and activation of caspase-9 and caspase-3, but not of caspase-8. In addition, thimerosal in a concentration-dependent manner inhibited the expression of XIAP, cIAP-1 but did not influence cIAP-2 expression. Furthermore, thimerosal enhanced intracellular reactive oxygen species and reduced intracellular glutathione (GSH). Finally, exogenous glutathione protected T cells from thimerosal-induced apoptosis by upregulation of XIAP and cIAP1 and by inhibiting activation of both caspase-9 and caspase-3. These data suggest that thimerosal induces apoptosis in T cells via mitochondrial pathway by inducing oxidative stress and depletion of GSH.

22. Mitochondrial mediated thimerosal-induced apoptosis in a human neuroblastoma cell line (SK-N-SH)
Humphrey ML, Cole MP, Pendergrass JC, Kiningham KK.
Neurotoxicology. 2005 Jun;26(3):407-16.

Environmental exposure to mercurials continues to be a public health issue due to their deleterious effects on immune, renal and neurological function. Recently the safety of thimerosal, an ethyl mercury-containing preservative used in vaccines, has been questioned due to exposure of infants during immunization. Mercurials have been reported to cause apoptosis in cultured neurons; however, the signaling pathways resulting in cell death have not been well characterized. Therefore, the objective of this study was to identify the mode of cell death in an in vitro model of thimerosal-induced neurotoxicity, and more specifically, to elucidate signaling pathways which might serve as pharmacological targets. Within 2 h of thimerosal exposure (5 microM) to the human neuroblastoma cell line, SK-N-SH, morphological changes, including membrane alterations and cell shrinkage, were observed. Cell viability, assessed by measurement of lactate dehydrogenase (LDH) activity in the medium, as well as the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, showed a time- and concentration-dependent decrease in cell survival upon thimerosal exposure. In cells treated for 24 h with thimerosal, fluorescence microscopy indicated cells undergoing both apoptosis and oncosis/necrosis. To identify the apoptotic pathway associated with thimerosal-mediated cell death, we first evaluated the mitochondrial cascade, as both inorganic and organic mercurials have been reported to accumulate in the organelle. Cytochrome c was shown to leak from the mitochondria, followed by caspase 9 cleavage within 8 h of treatment. In addition, poly(ADP-ribose) polymerase (PARP) was cleaved to form a 85 kDa fragment following maximal caspase 3 activation at 24 h. Taken together these findings suggest deleterious effects on the cytoarchitecture by thimerosal and initiation of mitochondrial-mediated apoptosis.

23. Thimerosal induces neuronal cell apoptosis by causing cytochrome c and apoptosis-inducing factor release from mitochondria
Yel L et al.
Int J Mol Med. 2005 Dec;16(6):971-7.

There is a worldwide increasing concern over the neurological risks of thimerosal (ethylmercury thiosalicylate) which is an organic mercury compound that is commonly used as an antimicrobial preservative. In this study, we show that thimerosal, at nanomolar concentrations, induces neuronal cell death through the mitochondrial pathway. Thimerosal, in a concentration- and time-dependent manner, decreased cell viability as assessed by calcein-ethidium staining and caused apoptosis detected by Hoechst 33258 dye. Thimerosal-induced apoptosis was associated with depolarization of mitochondrial membrane, generation of reactive oxygen species, and release of cytochrome c and apoptosis-inducing factor (AIF) from mitochondria to cytosol. Although thimerosal did not affect cellular expression of Bax at the protein level, we observed translocation of Bax from cytosol to mitochondria. Finally, caspase-9 and caspase-3 were activated in the absence of caspase-8 activation. Our data suggest that thimerosal causes apoptosis in neuroblastoma cells by changing the mitochondrial microenvironment.

24. Are toxic biometals destroying your children's future?
Drum DA.
Biometals. 2009 Oct;22(5):697-700. Epub 2009 Feb 11.
http://www.springerlink.com/content/7k502300609t72v4/

Cadmium, arsenic, lead, and mercury have been linked to autism, attention deficit disorder, mental retardation and death of children. Mercury in thimerosal found in many vaccines and flu shots contributes significantly to these problems. Decomposition of the thimerosal can produce more toxic compounds, either methylethylmercury or diethylmercury, in the body. These compounds have a toxicity level similar to dimethylmercury. Within the human body, a mitochondrial disorder may release the more toxic form of mercury internally. Young children and pregnant women must minimize internal exposure to the vaccines and flu shots containing mercury.

25. Developmental regression and mitochondrial dysfunction in a child with autism
Poling JS, Frye RE, Shoffner J, Zimmerman AW.
J Child Neurol. 2006 Feb;21(2):170-2.
{free online}
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2536523/?tool=pubmed

26. Mitochondrial dysfunction in autism spectrum disorders: a population-based study
Oliveira G et al.
Dev Med Child Neurol. 2005 Mar;47(3):185-9.

27. Mitochondrial disease in autism spectrum disorder patients: a cohort analysis
Weissman JR et al.
PLoS One. 2008;3(11):e3815. Epub 2008 Nov 26.
{free online}
http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0003815

28. Evidence of Mitochondrial Dysfunction in Autism and Implications for Treatment
Daniel A. Rossignol, J. Jeffrey Bradstreet
American Journal of Biochemistry and Biotechnology 4 (2): 208-217, 2008
{free online}
http://www.scipub.org/fulltext/ajbb/ajbb42208-217.pdf

29. Porphyrinuria in childhood autistic disorder: implications for environmental toxicity
Nataf R et al.
Toxicol Appl Pharmacol. 2006 Jul 15;214(2):99-108.

To address a possible environmental contribution to autism, we carried out a retrospective study on urinary porphyrin levels, a biomarker of environmental toxicity, in 269 children with neurodevelopmental and related disorders referred to a Paris clinic (2002-2004), including 106 with autistic disorder. Urinary porphyrin levels determined by high-performance liquid chromatography were compared between diagnostic groups including internal and external control groups. Coproporphyrin levels were elevated in children with autistic disorder relative to control groups. Elevation was maintained on normalization for age or to a control heme pathway metabolite (uroporphyrin) in the same samples. The elevation was significant (P < 0.001). Porphyrin levels were unchanged in Asperger's disorder, distinguishing it from autistic disorder. The atypical molecule precoproporphyrin, a specific indicator of heavy metal toxicity, was also elevated in autistic disorder (P < 0.001) but not significantly in Asperger's. A subgroup with autistic disorder was treated with oral dimercaptosuccinic acid (DMSA) with a view to heavy metal removal. Following DMSA there was a significant (P = 0.002) drop in urinary porphyrin excretion. These data implicate environmental toxicity in childhood autistic disorder.

30. A prospective study of mercury toxicity biomarkers in autistic spectrum disorders
Geier DA, Geier MR.
J Toxicol Environ Health A. 2007 Oct;70(20):1723-30.

Porphyrins are derivatives formed in the heme synthesis pathway and porphyrins afford a measure of xenobiotic exposure. The steps in the heme pathway most vulnerable to heavy metal inhibition are uroporphyrin decarboxylase (UROD) and coproporphyrinogen oxidase (CPOX) reactions. Mercury toxicity was associated with elevations in urinary coproporphyrin (cP), pentacarboxyporphyrin (5cxP), and precoproporphyrin (prcP) (also known as keto-isocoproporphyrin) levels. Two cohorts of autistic patients in the United States and France had urine porphyrin levels associated with mercury toxicity. A prospective study of urinary porphyrin testing at LabCorp (United States) and the Laboratoire Philippe Auguste (France) involving 71 autism spectrum disorder (ASD) patients, neurotypical sibling controls, and general population controls was undertaken. ASD patients had significant elevations in urinary levels of cP, 5cxP, and prcP relative to controls, and > 50% of ASD patients had urinary cP levels more than 2 standard deviations above the mean values for neurotypical sibling controls. Significant reductions in urinary 5cxP and cP levels were observed in ASD patients following chelation. A significant correlation was found between urinary porphyrins measured at LabCorp and those measured at the Laboratoire Philippe Auguste on individual ASD patients. The established developmental neurotoxicity attributed to mercury and biochemical/genomic evidence for mercury susceptibility/toxicity in ASDs indicates a causal role for mercury. Urinary porphyrin testing is clinically available, relatively inexpensive, and noninvasive. Porphyrins need to be routinely measured in ASDs to establish if mercury toxicity is a causative factor and to evaluate the effectiveness of chelation therapy.

31. Richard M. Kostrzewa
Editor-in-chief of Neutotoxicity Research
Department of Pharmacology, Quillen College of Medicine
East Tennessee State University, Johnson City, TN, USA
http://www.springer.com/biomed/neuroscience/journal/12640?detailsPage=aboutTheEditor

32. "Jeffrey R. Balser, M.D., Ph.D." <jeff.balser@vanderbilt.edu>
Associate Vice Chancellor for Health Affairs
Dean, School of Medicine
Vanderbilt University
http://www.vanderbilt.edu/

33. Harriet Wallberg-Henriksson <rektorssekreterare@ki.se>
President of Karolinska Institutet
Karolinska Institutet
http://info.ki.se/index_en.html

34. NIH: Center for Scientific Review
http://cms.csr.nih.gov/ContactCSR/


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