1. MERCURY TOXICITY: Dependence on Dose, Diet and Genetic Susceptibility DR. BOYD E. HALEY PROFESSOR AND CHAIR DEPARTMENT OF CHEMISTRY UNIVERSITY OF KENTUCKY

2. VISUALIZATION OF MERCURY EMITTING FROM A DENTAL AMALGAM From: www. uninformed consent.com From: www. uninformed consent.com David Kennedy’s IAOMT tape David Kennedy’s IAOMT tape

3. Measurement of Mercury Release from a Standard Dental Filling Amalgams of known weight and surface area were placed in a sealed test tube in 50 milliliters of H 2 O. H 2 O was changed daily and Hg level measured by a mercury cold vapor analyzer. Amalgams of known weight and surface area were placed in a sealed test tube in 50 milliliters of H 2 O. H 2 O was changed daily and Hg level measured by a mercury cold vapor analyzer. Without brushing the amalgams released 7.54  g Hg/cm 2 /day. Without brushing the amalgams released 7.54  g Hg/cm 2 /day. With two 30 second brushings with a medium toothbrush the amalgams released 45.5  g Hg/cm 2 /day, a six-fold increase. With two 30 second brushings with a medium toothbrush the amalgams released 45.5  g Hg/cm 2 /day, a six-fold increase. This confirms the earlier observation of Dr. Stock in 1926 in Zeitschr. Angew. Chem. 39, 461-466. This confirms the earlier observation of Dr. Stock in 1926 in Zeitschr. Angew. Chem. 39, 461-466.

4. Hg & THIMEROSAL DISPLAY ADDITIVE TOXICITIES TO NEURONS IN CULTURE.

5. MERCURY BIRTH HAIR LEVELS VS. AMALGAM FILLINGS IN AUTISTIC AND CONTROL GROUPS 0-3 Hair Hg level (mcg/g) Number of amalgams: Control: autistic ratio: 4-56-78-9>10 2.64 6.936.706.3217.91 N: 15 22293043 Autistic Control Data from A. Holmes, M. Blaxill & B. Haley, Int. J. of Toxicology v22, in press, 2003

6. ACTUAL VERSUS PREDICTED BIRTH HAIR MERCURY LEVELS Actual hair Hg level (ng/g) Hair Hg level= (5.60)+0.04(amalgam volume)+1.15(fish consumption)+0.03(vaccine): R2= 0.79 Predicted hair Hg level (mcg/g) Autistics Controls Data from A. Holmes, M. Blaxill & B. Haley, Int. J. Toxicology, in press, 2003

7. BIRTH-HAIR MERCURY BY SEVERITY OF AUTISM Hair Hg level (ppm) Mild Mean=0.71 n=27 Female Male Moderate Mean=0.46 n=43 Severe Mean=0.21 n=24 Data from Amy Holmes, Mark Blaxill & Boyd Haley, Int. J. Tocicology v22, in press, 2003.

8. CONTRAST BETWEEN BIRH HAIR Hg LEVELS AND BODY Hg LEVELS Autistic children have much lower Hg levels in their birth hair, yet Autistic children have much lower Hg levels in their birth hair, yet Numerous physicians have reported that autistic children carry a 5 to 6-fold higher mercury body burden than control children. Numerous physicians have reported that autistic children carry a 5 to 6-fold higher mercury body burden than control children. The obvious explanation is micro-mercuralism & genetic susceptibility to retention toxicity. The obvious explanation is micro-mercuralism & genetic susceptibility to retention toxicity. There is an obvious gender difference. This is explained by testosterone effects on T-toxicity. There is an obvious gender difference. This is explained by testosterone effects on T-toxicity.

9. ACROYDYNIA: Pink Disease Acroydynia was prevalent from about 1880 until 1940 affecting about 1 in 500 children. It was found to be caused by mercurous chloride (calomel) in baby teething powder. Removal of this teething powder from the market eliminated this disease which had some similarity to autism. Acroydynia was prevalent from about 1880 until 1940 affecting about 1 in 500 children. It was found to be caused by mercurous chloride (calomel) in baby teething powder. Removal of this teething powder from the market eliminated this disease which had some similarity to autism.

10. MOST IMPORTANT CONCLUSIONS THERE APPEARS TO BE A SUBSET OF THE POPULATION THAT CANNOT EFFECTIVELY EXCRETE MERCURY AND ARE AT GREATER RISK TO EXPOSURES TO MERCURY THAN ARE THE GENERAL POPULATION. THERE APPEARS TO BE A SUBSET OF THE POPULATION THAT CANNOT EFFECTIVELY EXCRETE MERCURY AND ARE AT GREATER RISK TO EXPOSURES TO MERCURY THAN ARE THE GENERAL POPULATION. THIS SUSCEPTIBILITY IS LIKELY DUE TO GENETIC DIFFERENCES, DIET, EXPOSURE TO OTHER TOXICANTS, ANTIBIOTICS, ETC. THIS SUSCEPTIBILITY IS LIKELY DUE TO GENETIC DIFFERENCES, DIET, EXPOSURE TO OTHER TOXICANTS, ANTIBIOTICS, ETC. DOES UNEXPLAINED LOW HAIR, NAIL, BLOOD OR URINE LEVELS APPEAR IN OTHER NEUROLOGICAL OR SYSTEMIC DISEASES? DOES UNEXPLAINED LOW HAIR, NAIL, BLOOD OR URINE LEVELS APPEAR IN OTHER NEUROLOGICAL OR SYSTEMIC DISEASES?

11. ELEVATED MERCURY IN IDIOPATHIC DILATED CARDIOMYOPATHY (IDCM). WHERE DOES Hg COME FROM? LEVELS ng/g HgSb Controls 8.0 1.5 IDCM178,400 19.260 This is a 22,300 fold increase in Hg! Frustaci et al., J. of American College of Cardiology, 33, (6) 1578, 1999. Controls were patients with valvular or ischemic heart disease. Athletic youth die of IDCM.

12. M. Bauman & K. Nelson Paper “In contrast, in the Sechylles study of >700 children, exposure was to marine fish only, and boys with higher levels of hair mercury performed better on some tests, including the Boston Naming Test and 2 tests of visual motor coordination.” “In contrast, in the Sechylles study of >700 children, exposure was to marine fish only, and boys with higher levels of hair mercury performed better on some tests, including the Boston Naming Test and 2 tests of visual motor coordination.” The above observation has lead some to dismiss Hg as being causal for any neurological problems in youth. The above observation has lead some to dismiss Hg as being causal for any neurological problems in youth. However, it is likely in light of the autistic observations that the boys with higher hair Hg levels were the best at excreting Hg or Methyl-Hg. However, it is likely in light of the autistic observations that the boys with higher hair Hg levels were the best at excreting Hg or Methyl-Hg.

13. CARDIOVASCULAR DISEASE AND MERCURY Two studies have found a correlation between cardiovascular disease and Hg. Guallar et al. NEJM v347,1747,2003 & Salonen et al. Atherosclerosis v148, 265, 2000. Two studies have found a correlation between cardiovascular disease and Hg. Guallar et al. NEJM v347,1747,2003 & Salonen et al. Atherosclerosis v148, 265, 2000. However, a Faeroe Island study found that blood pressure was increased in 7 year old children when the blood Hg levels were BELOW 10  g/liter, but not when it was higher! Sorensen et al. Epidemiology v10, 370, 1999 as reported by Clarkson et al. NEJM 2003. However, a Faeroe Island study found that blood pressure was increased in 7 year old children when the blood Hg levels were BELOW 10  g/liter, but not when it was higher! Sorensen et al. Epidemiology v10, 370, 1999 as reported by Clarkson et al. NEJM 2003. If ingestion rates of mercury were the same then the above results imply that low blood levels of Hg are caused by failure to excrete the mercury. If ingestion rates of mercury were the same then the above results imply that low blood levels of Hg are caused by failure to excrete the mercury.

14. Hg Levels in Human Brain Saxe et al, determined Hg levels in the brains of 101 human subjects, both AD and normals, mostly Catholic Nuns (Alzheimer’s Disease, Dental Amalgam and Mercury, JADA v130, p191-199, 1999). Saxe et al, determined Hg levels in the brains of 101 human subjects, both AD and normals, mostly Catholic Nuns (Alzheimer’s Disease, Dental Amalgam and Mercury, JADA v130, p191-199, 1999). The histogram in this paper showed 6 of 101 subjects with brain Hg levels above 200 ng/g wet weight (C=236, 248, 319: AD=394, 622, 698). This represents between 1.2 & 3.5 micromolar, or highly toxic levels of Hg in 6% of these subjects. At 100 ng/g Hg this increases to about 15% of subjects with highly toxic levels of brain mercury. The histogram in this paper showed 6 of 101 subjects with brain Hg levels above 200 ng/g wet weight (C=236, 248, 319: AD=394, 622, 698). This represents between 1.2 & 3.5 micromolar, or highly toxic levels of Hg in 6% of these subjects. At 100 ng/g Hg this increases to about 15% of subjects with highly toxic levels of brain mercury. Hg levels were double in the control olfactory tissue! Hg levels were double in the control olfactory tissue! Where does this Hg come from? Why is it elevated in certain individuals? This also appears to be an inability to excrete the mercury as the authors state Hg levels were not correlated with number of amalgam fillings. Where does this Hg come from? Why is it elevated in certain individuals? This also appears to be an inability to excrete the mercury as the authors state Hg levels were not correlated with number of amalgam fillings.

15. Hg levels in AD brain Ehmann, Markesbery, et al. Transactions of the American Nuclear Society, 41, 206-207, 1982 entitled “Brain Trace Element Studies of Aging and Disease by INAA” INAA= instrumental neutron activation analysis. Ehmann, Markesbery, et al. Transactions of the American Nuclear Society, 41, 206-207, 1982 entitled “Brain Trace Element Studies of Aging and Disease by INAA” INAA= instrumental neutron activation analysis. Tested for Ag, Br, Co, Cr, Cs, Fe, Hg, Rb, Sb, Sc, Se and Zn. Tested for Ag, Br, Co, Cr, Cs, Fe, Hg, Rb, Sb, Sc, Se and Zn. Quote: “ In the comparison of Alzheimer’s disease and control brains, the largest differences were observed for bromine and mercury, both being enriched in Alzheimer’s brains (p < 0.05).” Quote: “ In the comparison of Alzheimer’s disease and control brains, the largest differences were observed for bromine and mercury, both being enriched in Alzheimer’s brains (p < 0.05).”

16. Hg levels in hair & nails of AD Ehmann, Markesbery et al. Neurotoxicology 9(2)197-208. Trace Element Imbalances in Hair and Nails of Alzheimer’s Diseased Patients. Ehmann, Markesbery et al. Neurotoxicology 9(2)197-208. Trace Element Imbalances in Hair and Nails of Alzheimer’s Diseased Patients. Quote “The concentrations of 17 elements in the hair and nails of 180 Alzheimer’s disease (AD) and control subjects have been determined by instrumental neutron activation analysis (INAA). Quote “The concentrations of 17 elements in the hair and nails of 180 Alzheimer’s disease (AD) and control subjects have been determined by instrumental neutron activation analysis (INAA). “Mercury is decreased in the nail of AD subjects compared to controls” “Mercury is decreased in the nail of AD subjects compared to controls” “Perhaps one reason for the lowering of nail Hg in AD subjects could be the presumably lower exposure rate of the AD patients to environmental Hg. This would not explain the elevated brain Hg, however.” “Perhaps one reason for the lowering of nail Hg in AD subjects could be the presumably lower exposure rate of the AD patients to environmental Hg. This would not explain the elevated brain Hg, however.”

17. Hg in nails of AD subjects Ehman, Markesbery et al. Biological Trace Element Research, pp461-470. Editor:G.N. Schrauzer, 1990 by the Humana Press, Inc. Ehman, Markesbery et al. Biological Trace Element Research, pp461-470. Editor:G.N. Schrauzer, 1990 by the Humana Press, Inc. A Search for Longitudinal Variations in Trace Element Levels in Nails of Alzheimer’s disease patients. A Search for Longitudinal Variations in Trace Element Levels in Nails of Alzheimer’s disease patients. Quote “Mercury tended to decrease in nail with increasing age of patient, and with the duration and severity of the dementia.” Quote “Mercury tended to decrease in nail with increasing age of patient, and with the duration and severity of the dementia.”

18. Hg in nails of AD subjects Ehman, Markesbery et al. Biological Trace Element Research, pp461-470. Editor:G.N. Schrauzer, 1990 by the Humana Press, Inc. Ehman, Markesbery et al. Biological Trace Element Research, pp461-470. Editor:G.N. Schrauzer, 1990 by the Humana Press, Inc. A Search for Longitudinal Variations in Trace Element Levels in Nails of Alzheimer’s disease patients. A Search for Longitudinal Variations in Trace Element Levels in Nails of Alzheimer’s disease patients. Quote: “This decrease is counter to the elevated levels of Hg observed in AD brain, as compared to age-matched controls.” Quote: “This decrease is counter to the elevated levels of Hg observed in AD brain, as compared to age-matched controls.”

19. Hg Brain/Nail or Hair Ratios in AD versus Normal Subjects In a manner similar to that observed for autistic children the body burden of mercury in AD subjects appears to be elevated compared to controls whereas the mercury levels in the hair and nails was found to be significantly lower. In a manner similar to that observed for autistic children the body burden of mercury in AD subjects appears to be elevated compared to controls whereas the mercury levels in the hair and nails was found to be significantly lower. Perhaps these AD subjects could not excrete Hg as effectively as the normal subjects. Perhaps these AD subjects could not excrete Hg as effectively as the normal subjects. Do other experiments show any correlation between amalgam and brain Hg levels? Yes. Do other experiments show any correlation between amalgam and brain Hg levels? Yes.

20. Brain Hg levels versus Number Amalgam Surfaces From Dr. Magnus Nylander’s Research. The scatter in this graph, where high levels of Hg are observed with low amalgam surface count and low levels are observed with high amalgam count, and vice versa, is what would be expected if the individual subjects were differentially capable of excreting mercury. Similar results are seen doing similar analysis on fetal tissue and comparing Hg levels to birth mother’s amalgam count. The low Hg levels found in alcohol abusers also implies that diet may play a role in mercury retention.

21. Al:NEOMYCIN:TESTERONE EFFECTS + TESTOSTERONE SYNERGISTIC TOXICITIES on NEURONS IN CULTURE 50 NANOMOLAR DR. MARK LOVELL COLLABORATOR

22. Antibiotics, Milk and Hg Half-Life  In a study where rats were given high doses of oral antibiotics the half-life for excretion of mercury increased from 10 days to >100 days.  If the rats were also on a milk diet the excretion half-life increased to over 300 days. (Rowland, Archives of Environmental Health 1984: 39(6); 401-408)  It appears obvious that the cellular retention of Hg is dependent on genetics, diet and medical treatment. Increased cellular retention is an excretion problem.

23. Axonal Transport - A Process Essential for the Survival of Neurons Dendrite Microtubule, made of polymerized tubulin dimers. Membrane Bound Organelle Dynien Axon Kinesin

24. Autoradiogram of [ 32 P]8N 3 GTP Photolabeled Control & AD Brain Hippocampus Homogenates

25. EDTA Prevents Cd, Cu & Zn, But Not Hg, Inhibition of [ 32 P]8N 3 GTP Photolabeling of Brain ß-Tubulin

26. Partitioning of ß-Tubulin is Aberrant in Alzheimer’s Diseased Brain Homogenates NORMALS ALZHEIMERS ANTIBODY DETECTION OF BETA-TUBULIN

27. MERCURY AND ALZHEIMER’S DISEASE Exposure of neuroblastoma cells to 10 -9 molar mercury increases Tau phosphorylation and secretion of beta- amyloid. Both of these events occur in Alzheimer’s diseased brain. Amyloid plaque formation is the “diagnostic hallmark” of Alzheimer’s disease. Olivieri et al. J. Neurochemistry, 74, 231, 2000. Exposure of neuroblastoma cells to 10 -9 molar mercury increases Tau phosphorylation and secretion of beta- amyloid. Both of these events occur in Alzheimer’s diseased brain. Amyloid plaque formation is the “diagnostic hallmark” of Alzheimer’s disease. Olivieri et al. J. Neurochemistry, 74, 231, 2000. Exposure of cultured neurons to 10 -7 to 10 -10 molar mercury rapidly causes the stripping of tubulin from the neurofibrils forming the neurite processes leading to the formation of neurofibillary tangles, a “diagnostic hallmark” of Alzheimer’s disease. Leong et al. NeuroReports 12(4), 733, 2001 Exposure of cultured neurons to 10 -7 to 10 -10 molar mercury rapidly causes the stripping of tubulin from the neurofibrils forming the neurite processes leading to the formation of neurofibillary tangles, a “diagnostic hallmark” of Alzheimer’s disease. Leong et al. NeuroReports 12(4), 733, 2001 Therefore, Hg exposure can create these diagnostic hallmarks of Alzheimer’s Disease! Therefore, Hg exposure can create these diagnostic hallmarks of Alzheimer’s Disease!

28. HgEDTA Induces Aberrant [ 32 P]8N 3 GTP- ß-Tubulin Interactions Indicative of AD

29. EFFECT OF SEQUENTIAL AMALGAM EXTRACTION SOLUTIONS ON THE VIABILITY OF BRAIN TUBULIN Hours of Amalgam Soak % Active

30. PROTEINS INVOLVED IN Hg EXCRETION GLUTATHIONE (carries Hg through bilary transport, rapidly oxidized by excess Hg 2+. Note: Hg 2+ must go intracellular to interact with GSH, then be excreted for clearance in the bilary transport system of the liver. GLUTATHIONE (carries Hg through bilary transport, rapidly oxidized by excess Hg 2+. Note: Hg 2+ must go intracellular to interact with GSH, then be excreted for clearance in the bilary transport system of the liver. GLUTATHIONE TRANSFERASE, a recent gene product identified with AD risk. GLUTATHIONE TRANSFERASE, a recent gene product identified with AD risk. METALLOTHIONINE (Dr. W.Walsh’s work, regulated by estrogen and testosterone). This may explain gender susceptibility. METALLOTHIONINE (Dr. W.Walsh’s work, regulated by estrogen and testosterone). This may explain gender susceptibility. APO-E (apo-lipoprotein-E, a brain “housekeeping protein” that carries cholesterol from the brain through the CSF, through the blood-brain barrier for excretion by the liver bilary transport system.) This may explain additional excretion deficit. APO-E (apo-lipoprotein-E, a brain “housekeeping protein” that carries cholesterol from the brain through the CSF, through the blood-brain barrier for excretion by the liver bilary transport system.) This may explain additional excretion deficit.

31. Biochemical Differences Between the Three Most Common Apolipoprotein E Isoforms is Reflected in Hg Binding Capacity HIGH RISK FOR EARLY AD LOW RISK FOR EARLY AD 2 Mercury binding sites 1 Mercury binding site No Mercury binding sites

32. RELATIONSHIP TO NUMBER OF APO-E –SH GROUPS AND AGE OF AD ONSET APO-E  234  24 (>90)3 (>90)2 (80-90) 33 (>90)2 (80-90)1 (70-80) 42 (80-90)1 (70-80)0 (<70) RED = APO-E GENOTYPE COMBINATION YELLOW = NUMBER OF –SH GROUPS IN COMBINATION WHITE = APPROXIMATE AGE OF ONSET OF AD

33. M. Godfrey, et al. APO-E Geneotyping as a Potential Biomarker for Mercury Neurotoxicity. J. of Alzheimer’s Disease (June 2003) 400 subjects with a mean of 13.7 amalgams, with previously diagnosed neurological problems, were compared to 426 blood donors with regards to APO-E geneotype. 400 subjects with a mean of 13.7 amalgams, with previously diagnosed neurological problems, were compared to 426 blood donors with regards to APO-E geneotype. It was found that the symptomatic cohort was significantly elevated in percentages of 4/4 (3.5 vs 1.0 ) and low in the 2/2 (0.25 vs 1.4) in comparison to the blood donors. It was found that the symptomatic cohort was significantly elevated in percentages of 4/4 (3.5 vs 1.0 ) and low in the 2/2 (0.25 vs 1.4) in comparison to the blood donors. In mixed geneotypes 2/3 (9.5 vs 20) and 3/4 (30.8 vs 25) similar skewed ratios were found. In mixed geneotypes 2/3 (9.5 vs 20) and 3/4 (30.8 vs 25) similar skewed ratios were found. The 3/3 homozygote was found at 54.8 and 51.4 in the symptomatic and blood donor groups, respectively. The 3/3 homozygote was found at 54.8 and 51.4 in the symptomatic and blood donor groups, respectively.

34. Effect of Removal of Amalgams & Antioxidant Therapy on Patient Symptoms and Blood Hg Levels. U. Lindh et al., Neuroendocrinology Letters, v23, 459, 2002. Controls matched with test groups in age, gender & socio-economic conditions. They were subjectively healthy persons not suffering from any disease.

35. CONCLUSIONS WITH REGARD TO AD MOST, IF NOT ALL, ABERRANT BIOCHEMISTRY IN AD BRAIN CAN BE MIMICKED BY Hg. MOST, IF NOT ALL, ABERRANT BIOCHEMISTRY IN AD BRAIN CAN BE MIMICKED BY Hg. THE DIAGNOSTIC HALLMARKS OF AD BRAIN CAN BE PRODUCED BY Hg CONCENTRATIONS LOWER THAN REPORTED IN HUMAN BRAIN. THE DIAGNOSTIC HALLMARKS OF AD BRAIN CAN BE PRODUCED BY Hg CONCENTRATIONS LOWER THAN REPORTED IN HUMAN BRAIN. THE BIOLOGICAL PLAUSIBILITY OF Hg BEING CAUSAL IN AD IS MORE COMPLETE THAN THIMEROSAL CAUSATION OF AUTISM. THE BIOLOGICAL PLAUSIBILITY OF Hg BEING CAUSAL IN AD IS MORE COMPLETE THAN THIMEROSAL CAUSATION OF AUTISM. WHILE THERE HAS BEEN NO SIGNIFICANT EPIDEMIOLOGICAL STUDY ON Hg EXPOSURE AND AD, AND REASONABLE PEOPLE COULD DISAGREE ABOUT THE CAUSALITY, THERE IS NO DOUBT THAT EXPOSURE TO Hg WOULD EXACERBATE AD! WHILE THERE HAS BEEN NO SIGNIFICANT EPIDEMIOLOGICAL STUDY ON Hg EXPOSURE AND AD, AND REASONABLE PEOPLE COULD DISAGREE ABOUT THE CAUSALITY, THERE IS NO DOUBT THAT EXPOSURE TO Hg WOULD EXACERBATE AD!

36. CONCLUSION REGARDING AMALGAM FILLINGS DUE TO THE ENHANCEMENT OF Hg TOXICITY AND RETENTION BY FACTORS FOUND IN THE DIET, ANTIBIOTICS, OTHER TOXICANTS SUCH AS Pb 2+ & Cd 2+, AND GENETIC SUSCEPTIBILITY THERE CANNOT BE ANY REASONABLE PREDICITION OF A SAFE LEVEL OF MERCURY EXPOSURE WITHIN THE LEVELS PRODUCED BY HAVING AMALGAM FILLINGS. DUE TO THE ENHANCEMENT OF Hg TOXICITY AND RETENTION BY FACTORS FOUND IN THE DIET, ANTIBIOTICS, OTHER TOXICANTS SUCH AS Pb 2+ & Cd 2+, AND GENETIC SUSCEPTIBILITY THERE CANNOT BE ANY REASONABLE PREDICITION OF A SAFE LEVEL OF MERCURY EXPOSURE WITHIN THE LEVELS PRODUCED BY HAVING AMALGAM FILLINGS. MERCURY FROM THE BIRTH MOTHER’S AMALGAMS IS FOUND IN THE CHILD ON BIRTH. MERCURY FROM THE BIRTH MOTHER’S AMALGAMS IS FOUND IN THE CHILD ON BIRTH.