Teresa
Binstock
Researcher in Developmental & Behavioral Neuroanatomy
February 22, 2010
Start of autistic symptoms in infants parallels CDC's vaccination
schedule for infants and toddlers
Introduction:
As a scientific concept, "coincidence" is considered in a wiki entry
(1). Many parents of a child who regressed into autism in the days or
weeks following a vaccination have been told that the temporal
proximity of the vaccination(s) and the child's regression(s) was
merely coincidence (eg, 50). However, the fact that many observers
declare that autistic regressions after vaccinations are merely
coincidence neither proves nor disproves that vaccinations can induce
regression into autism or one of the other autism-spectrum disorders.
As considered herein, a growing body of peer-reviewed data suggests
that the notion of coincidence in regard to vaccinations and autism as
not as valid as proponents would prefer us to accept. Indeed, evidence
of an etiologically significant relationship between vaccinations and
autistic-regression is sufficient that large-N prospective studies of
vaccinated versus unvaccinated children are in order. Such studies
could be designed so as to demonstrate a likelihood of coincidence or
so as to indicate a likelihood of causal relationship between
vaccinations and autism.
New study: A newly published study documents
that, for most children who develop traits consistent with autism,
those traits tend to begin when the infant is 6 months old or somewhat
older (1). Whether cause or coincidence, the autism-onset timeline
reported by Ozonoff and colleagues (1) is remarkably similar to the
CDC's vaccination schedule for U.S. children (2). On February 18, 2010,
the study
and the vaccination
schedule were available free online and may still be. If so, the
two documents can be saved, perused, and compared.
When parents report that their child's autism or regression towards
autism began soon after the child had been vaccinated, experts often
state, "Coincidence". However, the invoking of "coincidence" in regard
to vaccinations and autism is a judgmental speculation not founded in
scientific data. Until large-N studies of vaccinated versus
unvaccinated children are enacted, using "coincidence" to explain
vaccination-associated regressions into autism is a faith-based
assertion which is void of supporting data.
The Ozonoff et al study is important. Its abstract merits
contemplation. The entire study merits perusal. In the abstract
hereinbelow, rhetoric that prompted two initial reactions is denoted by
asterisks.
Abstract:
Objective: To examine prospectively
the emergence of behavioral signs of autism in the first years of life
in infants at low and high risk for autism. Method: A prospective
longitudinal design was used to compare 25 infants later diagnosed with
an autism spectrum disorder (ASD) with 25 gender-matched low-risk
children later determined to have typical development. Participants
were evaluated at 6, 12, 18, 24, and 36 months of age. Frequencies of
gaze to faces, social smiles, and directed vocalizations were coded
from video and rated by examiners. Results: The frequency of gaze to
faces, shared smiles, and vocalizations to others were highly
comparable between groups at 6 months of age, but significantly
declining trajectories over time were apparent in the group later
diagnosed with ASD. Group differences were significant by 12 months of
age on most variables. Although repeated evaluation documented loss of
skills in most infants with ASD, most parents did not report a
regression in their child’s development [*]. Conclusions: These results suggest that behavioral signs of
autism are not present at birth, as once suggested by
Kanner[**], but emerge over time through a process of diminishment of
key social communication behaviors. More children may present with a
regressive course than previously thought, but parent report methods do
not capture this phenomenon well[*]. Implications for onset
classification systems and clinical screening are also
discussed. (1)
*[Comment] Parents are taught that individual children develop
at a rate different for each child, although group trends and average
stages have been described. Pediatricians often and correctly counsel
young parents that some delay is normal from some individual children.
This recommendation (in some cases) may mask parents' observations of
delay and thereby may minimize parental reports of a child's atypical
developmental progress. What Ozonoff et al reported regarding parental
observations seems not a fault of the parents.
**[Comment] Circa the
timing of this new study (2009-2010), the rate of autism is different
from the rate during Kanner's original
observations of a then-rare syndrome. At least some and perhaps many of
the autism cases Kanner described may have been present from birth and
thus may have been different from cases of regressive autism, as have
developed in recent decades, as observed by Ozonoff et al.
Not Necessarily Coincidence: The findings by Ozonoff et al (1) are remarkably parallel to
the CDC's timing of infant vaccinations (2). Thus, the new findings of
autism-onset timings are consistent with anecdotal reports from parents
who witnessed and later describe their child's regression toward autism
soon after one or several vaccination incidents. Indeed, several
factors described in peer-reviewed research articles suggest that
coincidence is unlikely in regard to many and perhaps most children who
regress soon after a vaccination. These several factors are considered
here:
Epidemiology:
Some epidemiological studies document neurologic sequelae to
vaccinations (eg, 3-9). Findings such as these are generally ignored by
experts who claim - often in major media - that no proof of harm
exists. Furthermore, reporting systems for vaccination-related adverse
events (NVICP and VAERS) exist because neurologic sequelae from
vaccinations have long been known (10-13).
Mechanisms:
Mechanisms by which thimerosal injures have been described (eg, 14-22),
as have mechanisms by which the MMR injures (eg, 3; see also 4).
Co-factors& models: A child in the U.S. is exposed to various environmental
factors associated with autism. As a result, the child's vaccinations
occur amid his or her body burden of various pollutants. These have
been found in amniotic fluid, cord blood, and breast milk (eg, 23-40).
Low levels of bisphenol A (BPA) affect human placenta cells (41).
Increased rates of autism have been found associated with various
airborne pollutants and with pesticides. (eg, 42-47).
One-pollutant models of autism causation differ from real life, wherein
many factors occur simultaneously. Similarly, vaccinations occur in a
context of other pollutants in the family's locale of residence and
pollutants within bodies (body burden, toxic load). Models,
experimental design, and statistical methods need incorporate
multi-pollutant exposures including toxicants in vaccines (eg, 51) and
also need incorporate the many-intra-body pollutants as the context in
which vaccinations occur. Focusing upon one pollutant per study can be
important but turns attention away from effects of combinations of
pollutants, many of which have become intra-body pollutants.
Genetics:
Peer-reviewed medical literature offers numerous studies describing
factors relevant to vaccinations and to the detoxification of vaccinal
and other environmental pollutants. Pubmed offers citations about
alleles that impair detoxification, that impair nutrient status, that
overly heighten immune responses. Other peer-reviewed studies describe
null alleles and other immune-related polymorphisms that would
adversely shape reactions to live viruses in certain vaccines. In
relation to thimerosal and glutathione, relevant genetics have been
described in studies led by Westphal G and by James SJ.
Changed schedule of
vaccinations: The increased number of
vaccinations imposed upon an infant, toddler, and young child (52)
remains untested in regard to safety. Some researchers and physicians
describe "vaccinosis" (53-54). A large-scale, long-term outcome study
of vaccinated children versus unvaccinated children remains sorely
needed.
The policy of vaccinating sick kids or
recently sick kids or not doing so: In
2000, when I, other co-authors of the autism-mercury hypothesis (55),
and several colleagues met with the FDA's Center for Biologics
Evaluation & Research (CBER) (56), I raised the glutathione-related
issue of not vaccinating sick or recently sick children. William
Miller, M.D., then acting director of the CBER, explained that not
vaccinating sick or recently sick children formerly had been the
officially recommended policy but, in order to increase vaccine
coverage, that policy had been changed to one of recommending that sick
and recently sick children be vaccinated. In my opinion and for reasons
related to glutathione depletion, this policy change has contributed to
a vaccination-related increase in autism and other autism-spectrum
disorders.
Into the future:
Vacc Unvacc study the only way to prove
"coincidence": When experts or others
invoke "coincidence" as a way of explaining the vaccination-autism
connection, we should keep in mind that one or more major, unbiased
vaccinated/unvaccinated studies is required to prove "coincidence".
Pre-screening via
advances in gene alleles and microarrays: The vaccination policy of one-size-fits-virtually-all for
specific vaccines and for the overall vaccine schedule needs be revised
by incorporation of more elaborate but efficient pre-screening of
infants. Microarray PCR technology can be used to identify infants most
likely to be injured by vaccinations or by multiple vaccinations in one
incident. Relevant genes would include but not be limited to weak
alleles in detox pathways or nutrient pathways, strong alleles in genes
related to inflammatory responses, and weak or null alleles in various
MHC genes related to immunity. Similar but not identical screening is
often used in refining cancer treatments. Infants deserve similar
consideration.
Ethics:
Matters of ethics are most perplexing.
Despite data indicating that thimerosal injures some children,
use of thimerosal in flu shots and some other vaccines means that
repeated injections of thimerosal are occurring. These injection are
happening despite data indicating that a small but important percentage
of individuals will be adversely affected.
Despite data showing the pre-screening before vaccinations could
identify the small percentage of children likely to be injured,
pre-screening seems not being developed.
Some have asked: Are we witnessing the enforcement of
intentionally iatrogenic medicine? Are cases of children recovered from
autism belittled if biomedical therapeutics were efficacious? Two
essays are among many addressing these important topics (57-58)
References:
1. A Prospective
Study of the Emergence of Early Behavioral Signs of
Autism
Sally Ozonoff et al.
J. Am. Acad. Child Adolesc. Psychiatry, 2010;49(3):258 –268.
available here
2. CDC vaccination
schedule, 2010
http://www.cdc.gov/vaccines/recs/schedules/downloads/child/2010/10_0-6yrs-schedule-pr.pdf
3. Acute
encephalopathy followed by permanent brain injury or death associated
with further attenuated measles vaccines: a review of claims submitted
to the National Vaccine Injury Compensation
Program.
Weibel RE, Caserta V, Benor DE, Evans G.
Division of Vaccine Injury Compensation, National Vaccine Injury
Compensation Program, Health Resources and Services Administration,
Public Health Service, Rockville, Maryland 20857, USA.
Pediatrics. 1998 Mar;101(3 Pt 1):383-7.
OBJECTIVE: To determine if there is evidence for a causal relationship
between acute encephalopathy followed by permanent brain injury or
death associated with the administration of further attenuated measles
vaccines(Attenuvax or Lirugen, Hoechst
Marion Roussel, Kansas City, MO), mumps vaccine(Mumpsvax, Merck and
Co, Inc, West Point, PA), or rubella vaccines (Meruvax or Meruvax II,
Merck and Co, Inc, West Point, PA), combined measles and rubella vaccine(M-R-Vax or M-R-Vax II, Merck and Co, Inc, West Point, PA), or
combined measles, mumps,
and rubella vaccine(M-M-R or M-M-R II,
Merck and Co, Inc, West Point, PA), the lead author reviewed claims
submitted to the National Vaccine Injury Compensation Program. METHODS:
The medical records of children who met the inclusion criteria of
receiving the first dose of these vaccines between 1970 and 1993 and
who developed such an encephalopathy with no determined cause within 15
days were identified and analyzed. RESULTS: A total of 48 children,
ages 10 to 49 months, met the inclusion criteria after receiving
measles vaccine, alone or in combination. Eight children died, and the
remainder had mental regression and retardation, chronic seizures,
motor and sensory deficits, and movement disorders. The onset of
neurologic signs or symptoms occurred with a nonrandom, statistically
significant distribution of cases on days 8 and 9. No cases were identified after the
administration of monovalent mumps or rubella
vaccine. CONCLUSIONS: This clustering suggests that a causal
relationship between measles vaccine and encephalopathy may exist as a
rare complication of measles immunization.
4. [An excellent and thorough review]
Measles, mumps,
rubella vaccine: through a glass, darkly.
Wakefield AJ, Montgomery SM.
Adverse Drug React Toxicol Rev. 2000 Dec;19(4):265-83; reviewer
comments 284-92.
5. [FOIA-based summary of CDC data compiled by Verstraeten et al 1999.
The data became diluted and published as Verstraeten et al 2003]
Generation
Zero{Analysis of CDC's 1999
thimerosal findings}
Blaxill M, Safeminds 2004
http://www.safeminds.org/research/library/GenerationZeroPowerPoint.pdf
6. A comparative
evaluation of the effects of MMR immunization and mercury doses from
thimerosal-containing childhood vaccines on the population prevalence
of autism.
Geier DA, Geier MR.
Med Sci Monit. 2004 Mar;10(3):PI33-9. Epub 2004 Mar 1.
http://www.medscimonit.com/fulltxt.php?ICID=11608
{free download if for personal use}
BACKGROUND: The purpose of the study was to evaluate the effects of MMR
immunization and mercury from thimerosal-containing childhood vaccines
on the prevalence of autism. MATERIAL/METHODS: Evaluations of the
Biological Surveillance Summaries of the Centers for Disease Control
and Prevention (CDC), the U.S. Department of Education datasets, and
the CDC's yearly live birth estimates were undertaken RESULTS: It was
determined that there was a close correlation between mercury doses
from thimerosal--containing childhood vaccines and the prevalence of
autism from the late 1980s through the mid-1990s. In contrast, there
was a potential correlation between the number of primary pediatric
measles-containing vaccines administered and the prevalence of autism
during the 1980s. In addition, it was found that there were
statistically significant odds ratios for the development of autism
following increasing doses of mercury from thimerosal-containing
vaccines (birth cohorts: 1985 and 1990-1995) in comparison to a
baseline measurement (birth cohort: 1984). The contribution of
thimerosal from childhood vaccines (>50% effect) was greater than
MMR vaccine on the prevalence of autism observed in this study.
CONCLUSIONS: The results of this study agree with a number of
previously published studies. These studies have shown that there is biological plausibility
and epidemiological evidence showing a direct relationship between
increasing doses of mercury from thimerosal-containing vaccines and
neurodevelopmental disorders, and measles-containing vaccines and
serious neurological disorders. It is recommended that thimerosal be
removed from all vaccines, and additional research be undertaken to
produce a MMR vaccine with an improved safety
profile.
7. Thimerosal
exposure in infants and neurodevelopmental disorders: an assessment of
computerized medical records in the Vaccine Safety
Datalink.
Young HA, Geier DA, Geier MR.
The George Washington University School of Public Health and Health
Services
J Neurol Sci. 2008 Aug 15;271(1-2):110-8.
$ http://linkinghub.elsevier.com/retrieve/pii/S0022-510X%2808%2900157-3
The study evaluated possible associations between neurodevelopmental
disorders (NDs) and exposure to mercury (Hg) from Thimerosal-containing
vaccines (TCVs) by examining the automated Vaccine Safety Datalink
(VSD). A total of 278,624 subjects were identified in birth cohorts
from 1990-1996 that had received their first oral polio vaccination by
3 months of age in the VSD. The birth cohort prevalence rate of
medically diagnosed International Classification of Disease, 9th
revision (ICD-9) specific NDs and control outcomes were calculated.
Exposures to Hg from TCVs were calculated by birth cohort for specific
exposure windows from birth-7 months and birth-13 months of age.
Poisson regression analysis was used to model the association between
the prevalence of outcomes and Hg doses from TCVs. Consistent significantly increased rate
ratios were observed for autism, autism spectrum disorders, tics,
attention deficit disorder, and emotional disturbances with Hg exposure
from TCVs. [A finding consistent with
Verstraeten et al 1999] By contrast, none of the control outcomes had
significantly increased rate ratios with Hg exposure from TCVs. Routine
childhood vaccination should be continued to help reduce the morbidity
and mortality associated with infectious diseases, but efforts should
be undertaken to remove Hg from vaccines. Additional studies should be
conducted to further evaluate the relationship between Hg exposure and
NDs.
8. 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.
9. 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.
10. NVICP
http://www.hrsa.gov/vaccinecompensation/default.htm
11. The Clinical
Impact of Adverse Event Reporting
FDA, 1996.
http://www.fda.gov/downloads/Safety/MedWatch/UCM168505.pdf
12. Vaccine Adverse
Event Reporting System (VAERS)
HHS
http://vaers.hhs.gov/index
13. Post-marketing
surveillance for adverse events after vaccination: the national Vaccine
Adverse Event Reporting System (VAERS)
FDA, 1998.
http://www.fda.gov/downloads/Safety/MedWatch/UCM168497.pdf
14. 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.
{free online}
http://www.nature.com/mp/journal/v9/n4/pdf/4001476a.pdf
Methylation events play a critical role in the ability of growth
factors to promote normal development. Neurodevelopmental toxins, such
as ethanol and heavy metals, interrupt growth factor signaling, raising
the possibility that they might exert adverse effects on methylation.
We found that insulin-like growth factor-1 (IGF-1)- and
dopamine-stimulated methionine synthase (MS) activity and
folate-dependent methylation of phospholipids in SH-SY5Y human
neuroblastoma cells, via a PI3-kinase- and MAP-kinase-dependent
mechanism. The stimulation of this pathway increased DNA methylation,
while its inhibition increased methylation-sensitive gene expression.
Ethanol potently interfered with IGF-1 activation of MS and blocked its
effect on DNA methylation, whereas it did not inhibit the effects of
dopamine. Metal ions potently affected IGF-1 and dopamine-stimulated MS
activity, as well as folate-dependent phospholipid methylation: Cu(2+)
promoted enzyme activity and methylation, while Cu(+), Pb(2+), Hg(2+)
and Al(3+) were inhibitory. The ethylmercury-containing preservative
thimerosal inhibited both IGF-1- and dopamine-stimulated methylation
with an IC(50) of 1 nM and eliminated MS activity. Our findings outline
a novel growth factor signaling pathway that regulates MS activity and
thereby modulates methylation reactions, including DNA methylation. The
potent inhibition of this pathway by ethanol, lead, mercury, aluminum
and thimerosal suggests that it may be an important target of
neurodevelopmental toxins.
15. Thimerosal
induces DNA breaks, caspase-3 activation, membrane damage, and cell
death in cultured human neurons and
fibroblasts.
Baskin DS, Ngo H, Didenko VV.
Toxicol Sci. 2003 Aug;74(2):361-8. Epub 2003 May 28.
http://toxsci.oxfordjournals.org/cgi/content/full/74/2/361
Thimerosal is an organic mercurial compound used as a preservative in
biomedical preparations. Little is known about the reactions of human
neuronal and skin cells to its micro- and nanomolar concentrations,
which can occur after using thimerosal-containing products. A useful
combination of fluorescent techniques for the assessment of thimerosal
toxicity is introduced. Short-term thimerosal toxicity was investigated
in cultured human cerebral cortical neurons and in normal human
fibroblasts. Cells were incubated with 125-nM to 250-microM
concentrations of thimerosal for 45 min to 24 h. A 4',
6-diamidino-2-phenylindole dihydrochloride (DAPI) dye exclusion test
was used to identify nonviable cells and terminal transferase-based
nick-end labeling (TUNEL) to label DNA damage. Detection of active
caspase-3 was performed in live cell cultures using a cell-permeable
fluorescent caspase inhibitor. The morphology of fluorescently labeled
nuclei was analyzed. After 6 h of incubation, the thimerosal toxicity
was observed at 2 microM based on the manual detection of the
fluorescent attached cells and at a 1-microM level with the more
sensitive GENios Plus Multi-Detection Microplate Reader with Enhanced
Fluorescence. The lower limit did not change after 24 h of incubation.
Cortical neurons demonstrated higher sensitivity to thimerosal compared
to fibroblasts. The first sign of toxicity was an increase in membrane
permeability to DAPI after 2 h of incubation with 250 microM
thimerosal. A 6-h incubation resulted in failure to exclude DAPI,
generation of DNA breaks, caspase-3 activation, and development of
morphological signs of apoptosis. We demonstrate that thimerosal in
micromolar concentrations rapidly induce membrane and DNA damage and
initiate caspase-3-dependent apoptosis in human neurons and
fibroblasts. We conclude that a proposed combination of fluorescent
techniques can be useful in analyzing the toxicity of thimerosal.
16. 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.
17. 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.
18. 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.
19. Thimerosal
neurotoxicity is associated with glutathione depletion: protection with
glutathione precursors.
James SJ, Slikker W 3rd, Melnyk S, New E, Pogribna M, Jernigan S.
Neurotoxicology. 2005 Jan;26(1):1-8.
Thimerosol is an antiseptic containing 49.5% ethyl mercury that has
been used for years as a preservative in many infant vaccines and in
flu vaccines. Environmental methyl mercury has been shown to be highly
neurotoxic, especially to the developing brain. Because mercury has a
high affinity for thiol (sulfhydryl (-SH)) groups, the thiol-containing
antioxidant, glutathione (GSH), provides the major intracellular
defense against mercury-induced neurotoxicity. Cultured neuroblastoma
cells were found to have lower levels of GSH and increased sensitivity
to thimerosol toxicity compared to glioblastoma cells that have higher
basal levels of intracellular GSH. Thimerosal-induced cytotoxicity was
associated with depletion of intracellular GSH in both cell lines.
Pretreatment with 100 microM glutathione ethyl ester or
N-acetylcysteine (NAC), but not methionine, resulted in a significant
increase in intracellular GSH in both cell types. Further, pretreatment
of the cells with glutathione ethyl ester or NAC prevented cytotoxicity
with exposure to 15 microM Thimerosal. Although Thimerosal has been
recently removed from most children's vaccines, it is still present in
flu vaccines given to pregnant women, the elderly, and to children in
developing countries. The potential protective effect of GSH or NAC
against mercury toxicity warrants further research as possible adjunct
therapy to individuals still receiving Thimerosal-containing
vaccinations.
20. Cellular and
mitochondrial glutathione redox imbalance in lymphoblastoid cells
derived from children with autism.
James SJ, Rose S, Melnyk S, Jernigan S, Blossom S, Pavliv O, Gaylor
DW.
FASEB J. 2009 Aug;23(8):2374-83. Epub 2009 Mar 23.
$ http://www.fasebj.org/cgi/content/full/23/8/2374
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.
21. Evaluation of
cytotoxicity attributed to thimerosal on murine and human kidney
cells.
Park EK, Mak SK, Kültz D, Hammock BD.
J Toxicol Environ Health A. 2007 Dec;70(24):2092-5.
Renal inner medullary collecting duct cells (mIMCD3) and human
embryonic kidney cells (HEK293) were used for cytoscreening of
thimerosal and mercury chloride (HgCl2). Thimerosal and HgCl2 acted in
a concentration-dependent manner. In mIMCD3 cells the 24-h LC50 values
for thimerosal, thiosalicylic acid, 2,2-dithiosalicylic acid, and
2-sulfobenzoic acid were 2.9, 2200, >1000, and >10,000 microM,
respectively. The 24-h LC50 value for HgCl2 in mIMCD3 cells was 40
microM. In HEK293 cells, the 24-h LC50 value for thimerosal was 9.5
microM. These data demonstrate that the higher cytotoxicity produced by
thimerosal on renal cells with respect to similar compounds without Hg
may be related to this metal content. The present study also
establishes mIMCD3 cells as a valuable model for evaluation of
cytotoxicity of nephrotoxic compounds.
22. Neonatal
administration of a vaccine preservative, thimerosal, produces lasting
impairment of nociception and apparent activation of opioid system in
rats
Olczak M, Duszczyk M, Mierzejewski P, Majewska MD.
Brain Res. 2009 Dec 8;1301:143-51. Epub 2009 Sep 9.
Thimerosal (THIM), an organomercury preservative added to many child
vaccines is a suspected factor in pathogenesis of neurodevelopmental
disorders. We examined the pharmacokinetics of Hg in the brain, liver
and kidneys after i.m. THIM injection in suckling rats and we tested
THIM effect on nociception. THIM solutions were injected to Wistar and
Lewis rats in a vaccination-like mode on PN days 7, 9, 11 and 15 in
four equal doses. For Wistar rats these were: 12, 48, 240, 720, 1440,
2160, 3000 microg Hg/kg and for Lewis: 54, 216, 540 and 1080 microg
Hg/kg. Pharmacokinetic analysis revealed that Hg from THIM injections
accumulates in the rat brain in significant amounts and remains there
longer than 30 days after the injection. At the 6th week of age animals
were examined for pain sensitivity using the hot plate test. THIM
treated rats of both strains and sexes manifested statistically
significantly elevated pain threshold (latency for paw licking,
jumping) on a hot plate (56 degrees C). Wistar rats were more sensitive
to this effect than Lewis rats. Protracted THIM-induced hypoalgesia was
reversed by naloxone (5 mg/kg, i.p.) injected before the hot plate
test, indicative of involvement of endogenous opioids. This was
confirmed by augmented catalepsy after morphine (2.5 mg/kg, s.c.)
injection. Acute THIM injection to 6-week-old rats also produced
hypoalgesia, but this effect was transient and was gone within 14 days.
Present findings show that THIM administration to suckling or adult
rats impairs sensitivity to pain, apparently due to activation the
endogenous opioid system.
23. Detection of
endocrine disrupting chemicals in samples of second trimester human
amniotic fluid
Foster W, Chan S, Platt L, Hughes C.
J Clin Endocrinol Metab. 2000 Aug;85(8):2954-7.
24. Third-trimester
amniotic fluid metal levels associated with
preeclampsia
Dawson EB, Evans DR, Nosovitch J.
Arch Environ Health. 1999 Nov-Dec;54(6):412-5.
25. Polychlorinated
biphenyls in human amniotic fluid
Rao CV, Banerji AS.
Bull Environ Contam Toxicol. 1988 Dec;41(6):798-801.
26. Mercury in human
amniotic fluid
Suzuki T, Takemoto T, Shishido S, Kani K.
Scand J Work Environ Health. 1977 Mar;3(1):32-5.
27. Maternal and cord
blood levels of organochlorine pesticides: association with preterm
labor
Pathak R, Ahmed RS, Tripathi AK, Guleria K, Sharma CS, Makhijani SD,
Banerjee BD.
Clin Biochem. 2009 May;42(7-8):746-9.
28. Combined analysis
of prenatal (maternal hair and blood) and neonatal (infant hair, cord
blood and meconium) matrices to detect fetal exposure to environmental
pesticides
Ostrea EM Jr, Bielawski DM, Posecion NC Jr, Corrion M, Villanueva-Uy E,
Bernardo RC, Jin Y, Janisse JJ, Ager JW.
Environ Res. 2009 Jan;109(1):116-22. Epub 2008 Nov 18.
29. Association of
thyroid hormone concentrations with levels of organochlorine compounds
in cord blood of neonates
Maervoet J, Vermeir G, Covaci A, Van Larebeke N, Koppen G, Schoeters G,
Nelen V, Baeyens W, Schepens P, Viaene MK.
Environ Health Perspect. 2007 Dec;115(12):1780-6.
30. Placental
p,p'-dichlorodiphenyldichloroethylene and cord blood immune
markers
Brooks K, Hasan H, Samineni S, Gangur V, Karmaus W.
Pediatr Allergy Immunol. 2007 Nov;18(7):621-4
31. Cadmium, lead,
and selenium in cord blood and thyroid hormone status of
newborns
Iijima K, Otake T, Yoshinaga J, Ikegami M, Suzuki E, Naruse H, Yamanaka
T, Shibuya N, Yasumizu T, Kato N.
Biol Trace Elem Res. 2007 Oct;119(1):10-8.
32. Mercury levels in
cord blood and meconium of healthy newborns and venous blood of their
mothers: clinical, prospective cohort study
Unuvar E, Ahmadov H, Kiziler AR, Aydemir B, Toprak S, Ulker V, Ark
C.
Sci Total Environ. 2007 Mar 1;374(1):60-70. Epub 2007 Jan 26.
33. Distribution of
PCDDs/PCDFs and Co-PCBs in human maternal blood, cord blood, placenta,
milk, and adipose tissue: dioxins showing high toxic equivalency factor
accumulate in the placenta
Suzuki G, Nakano M, Nakano S.
Biosci Biotechnol Biochem. 2005 Oct;69(10):1836-47.
34. Detection of
prenatal exposure to several classes of environmental toxicants and
their metabolites by gas chromatography-mass spectrometry in maternal
and umbilical cord blood
Corrion ML, Ostrea EM Jr, Bielawski DM, Posecion NC Jr, Seagraves
JJ.
J Chromatogr B Analyt Technol Biomed Life Sci. 2005 Aug
5;822(1-2):221-9.
35. Human exposure to
endocrine disruptors and breast milk
Stefanidou M, Maravelias C, Spiliopoulou C.
Endocr Metab Immune Disord Drug Targets. 2009 Sep;9(3):269-76.
36. Relationship
between the concentrations of polychlorinated dibenzo-p-dioxins,
polychlorinated dibenzofurans, and polychlorinated biphenyls in
maternal blood and those in breast milk
Todaka T et al.
Chemosphere. 2010 Jan;78(2):185-92.
37. Levels and
temporal trends of chlorinated pesticides, polychlorinated biphenyls
and brominated flame retardants in individual human breast milk samples
from Northern and Southern Norway
Polder A, Thomsen C, Lindström G, Løken KB, Skaare JU.
Chemosphere. 2008 Aug;73(1):14-23. Epub 2008 Jul 23.
38. Polybrominated
diphenyl ethers and organochlorine pesticides in human breast milk from
Massachusetts, USA
Johnson-Restrepo B, Addink R, Wong C, Arcaro K, Kannan K.
J Environ Monit. 2007 Nov;9(11):1205-12.
39. Polybrominated
diphenyl ethers (PBDEs) and polychlorinated biphenyls (PCBs) in breast
milk from the Pacific Northwest
She J, Holden A, Sharp M, Tanner M, Williams-Derry C, Hooper K.
Chemosphere. 2007 Apr;67(9):S307-17.
40. Different levels
of polybrominated diphenyl ethers (PBDEs) and chlorinated compounds in
breast milk from two U.K. Regions
Kalantzi OI, Martin FL, Thomas GO, Alcock RE, Tang HR, Drury SC,
Carmichael PL, Nicholson JK, Jones KC.
Environ Health Perspect. 2004 Jul;112(10):1085-91.
41. Toxic effects of
low doses of Bisphenol-A on human placental
cells
Nora Benachoura and Aziz Aris
Toxicology and Applied Pharmacology Volume 241, Issue 3, 15 December
2009, Pages 322-328
$ http://tinyurl.com/y9wqhn5
Humans are exposed daily to a great number of xenobiotics and their
metabolites present as pollutants. Bisphenol-A (BPA) is extensively
used in a broad range of products including baby bottles, food-storage
containers, medical equipment, and consumer electronics. Thus, BPA is
the most common monomer for polycarbonates intended for food contact.
Levels of this industrial product are found in maternal blood, amniotic
fluid, follicular fluid, placental tissue, umbilical cord blood, and
maternal urine. In this study, we investigated toxic effects of BPA
concentrations close to levels found in serum of pregnant women on
human cytotrophoblasts (CTB). These cells were isolated from fresh
placentas and exposed to BPA for 24 h. Our results showed that very low
doses of BPA induce apoptosis (2 to 3 times) as assessed using M30
antibody immunofluorescent detection, and necrosis (1.3 to 1.7 times)
as assessed through the cytosolic Adenylate Kinase (AK) activity after
cell membrane damage. We also showed that BPA increased significantly
the tumor-necrosis factor alpha (TNF-α) gene expression and
protein excretion as measured by real-time RT-PCR and ELISA luminescent
test, respectively. Moreover, we observed that induction of AK
activation and TNF-α gene expression require lower levels of BPA
than apoptosis or TNF-α protein excretion. Our findings suggest
that exposure of placental cells to low doses of BPA may cause
detrimental effects, leading in
vivoto adverse pregnancy outcomes such as
preeclampsia, intrauterine growth restriction, prematurity and
pregnancy loss.
42. Environmental
mercury release, special education rates, and autism disorder: an
ecological study of Texas
Palmer RF, Blanchard S, Stein Z, Mandell D, Miller C.
Health Place. 2006 Jun;12(2):203-9.
43. Autism spectrum
disorders in relation to distribution of hazardous air pollutants in
the san francisco bay area
Windham GC, Zhang L, Gunier R, Croen LA, Grether JK.
Environ Health Perspect. 2006 Sep;114(9):1438-44.
44. Maternal
residence near agricultural pesticide applications and autism spectrum
disorders among children in the California Central
Valley
Roberts EM, English PB, Grether JK, Windham GC, Somberg L, Wolff C.
Environ Health Perspect. 2007 Oct;115(10):1482-9.
45. Proximity to
point sources of environmental mercury release as a predictor of autism
prevalence
Palmer RF, Blanchard S, Wood R.
Health Place. 2009 Mar;15(1):18-24.
46. Ockham's Razor
and autism: the case for developmental neurotoxins contributing to a
disease of neurodevelopment
DeSoto MC.
Neurotoxicology. 2009 May;30(3):331-7.
47. Autism Spectrum
Disorders and Identified Toxic Land Fills: Co-Occurrence Across
States
Xue Ming et al.
Environmental Health Insights 2008:2 55–59 [free online]
http://www.la-press.com/redirect_file.php?fileId=1420&filename=EHI-2-Ming-et-al&fileType=pdf
Abstract: It is believed that gene by environmental interactions
contribute to the pathogenesis of autism spectrum disorders (ASD). We
hypothesize that ASD are associated with early and repeated exposures
to any of a number of toxicants or mixtures of toxicants. It is the
cumulative effects of these repeated exposures acting upon genetically
susceptible individuals that lead to the phenotypes of ASD. We report
our initial observations of a considerable overlap of identified toxic
landfi lls in the State of New Jersey and the residence of an ASD
cohort, and a correlation between the identified toxic Superfund sites
on each U.S. state and the total number of diagnosed cases of ASD in
those states. The residence of 495 ASD patients in New Jersey by zip
code and the toxic landfill sites were plotted on a map of Northern New
Jersey. The area of highest ASD cases coincides with the highest
density of toxic landfill sites while the area with lowest ASD cases
has the lowest density of toxic landfi ll sites. Furthermore, the
number of toxic Superfund sites and autism rate across 49 of the 50
states shows a statistically signifi cant correlation (i.e. the number
of identified superfund sites correlates with the rate of autism per
1000 residents in 49 of the states (p = 0.015; excluding the state of
Oregon). These significant observations call for further organized
studies to elucidate possible role(s) of environmental toxicants
contributing to the pathogenesis of ASD.
48. Center for
Biologics Evaluation & Research(CBER)
http://www.fda.gov/BiologicsBloodVaccines/default.htm
49. Coindidence
http://en.wikipedia.org/wiki/Coincidence
50. Early Diagnosis
of Autism – Implications for the Vaccine
Hypothesis
Steven Novella
http://www.theness.com/neurologicablog/?p=184
51. Vaccine ingredients
http://www.sailhome.org/Concerns/Vaccines.html
52. Lists of Childhood Vaccinations, 1983, 2007
http://www.generationrescue.org/pdf/cdc_comparison.pdf
53. Vaccination and
autoimmunity-'vaccinosis': a dangerous
liaison?
Shoenfeld Y, Aron-Maor A.
J Autoimmun. 2000 Feb;14(1):1-10.
$ http://tinyurl.com/yfjntq
The question of a connection between vaccination and autoimmune illness
(or phenomena) is surrounded by controversy. A heated debate is going
on regarding the causality between vaccines, such as measles and
anti-hepatitis B virus (HBV), and multiple sclerosis (MS). Brain
antibodies as well as clinical symptoms have been found in patients
vaccinated against those diseases. Other autoimmune illnesses have been
associated with vaccinations. Tetanus toxoid, influenza vaccines, polio
vaccine, and others, have been related to phenomena ranging from
autoantibodies production to full-blown illness (such as rheumatoid
arthritis (RA)). Conflicting data exists regarding also the connection
between autism and vaccination with measles vaccine. So far only one
controlled study of an experimental animal model has been published, in
which the possible causal relation between vaccines and autoimmune
findings has been examined: in healthy puppies immunized with a variety
of commonly given vaccines, a variety of autoantibodies have been
documented but no frank autoimmune illness was recorded. The findings
could also represent a polyclonal activation (adjuvant reaction). The
mechanism (or mechanisms) of autoimmune reactions following
immunization has not yet been elucidated. One of the possibilities is
molecular mimicry; when a structural similarity exists between some
viral antigen (or other component of the vaccine) and a self-antigen.
This similarity may be the trigger to the autoimmune reaction. Other
possible mechanisms are discussed. Even though the data regarding the
relation between vaccination and autoimmune disease is conflicting, it
seems that some autoimmune phenomena are clearly related to
immunization (e.g. Guillain-Barre syndrome). The issue of the risk of
vaccination remains a philosophical one, since to date the advantages
of this policy have not been refuted, while the risk for autoimmune
disease has not been irrevocably proved. We discuss the pros and cons
of this issue (although the temporal relationship (i.e. always 2-3
months following immunization) is impressive)
54. Vaccines and
Autoimmune Diseases of the Adult
Hedi Orbach et al.
Copyright 2010, Discovery Medicine. All Rights Reserved.
http://www.discoverymedicine.com/HediOrbach/2010/02/04/vaccinesandautoimmunediseasesoftheadult/
Abstract: Infectious agents contribute to the environmental factors
involved in the development of autoimmune diseases possibly through
molecular mimicry mechanisms. Hence, it is feasible that vaccinations
may also contribute to the mosaic of autoimmunity. Evidence for the
association of vaccinations and the development of these diseases is
presented in this review. Infrequently reported postvaccination
autoimmune diseases include systemic lupus erythematosus, rheumatoid
arthritis, inflammatory myopathies, multiple sclerosis, GuillainBarré
syndrome, and vasculitis. In addition, we will discuss macrophagic
myofasciitis, aluminum containing vaccines, and the recent evidence for
autoimmunity following human papilloma virus vaccine.
55. Autism: a novel
form of mercury poisoning
Bernard S, Enayati A, Redwood L, Roger H, Binstock T.
Med Hypotheses. 2001 Apr;56(4):462-71.
56. Center for
Biologics Evaluation & Research(CBER)
http://www.fda.gov/BiologicsBloodVaccines/default.htm
57. Iatrogenecists
excel in the practice of iatrogenic medicine
Teresa Binstock, Jan 21, 2010
58. The
Phoenicians: Autism Recovery Denial, Drug Profits and the Media’s Flat
Earth
Adriana Gamondes, February 18, 2010
Contact Teresa Binstock by email
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