Mercury Biochemistry and its Relationship to Neurological Diseases

Mercury Biochemistry and its Relationship to Neurological Diseases Dr. Boyd Haley Professor Department of Chemistry University of Kentucky May 2007

VISUALIZATION OF MERCURY EMITTING FROM A DENTAL AMALGAM • From: www. uninformed concent. com • David Kennedy’s IAOMT tape IN SPITE OF THE OBVIOUS EMISSION OF Hg VAPORS FROM DENTAL AMALGAM THE FDA HAS STEADFASTL Y REFUSED TO TEST THEM FOR SAFETY!

IAOMT AMALGAM STUDY PROCEDURE • Nine dentists across the USA volunteered to make 10 cylindrical, one spill amalgams in a provided plexiglass mold. • The IAOMT provided new amalgam kits directly from the manufacturers to each dentist. • The amalgams in the molds were sent to Dr. Haley at the University of Kentucky for Hg analysis. • The amalgams were allowed to age for over one month to eliminate any surface mercury. • The amalgams were placed in 10 ml of distilled water which was changed daily. • Aliquots of this water were removed at days indicated analyzed for mercury content.

MA 2000 Nippon Direct Mercury Analyzer

DENTISTS BRAND DAY 1 DAY 4 DAY 8 DAY 1 1 BASCIANO Valiant 9. 921 9. 677 9. 580 9. 463 8. 700 8. 873 9. 392 9. 311 9. 751 9. 262 8. 886 8. 202 8. 074 8. 014 9. 563 10. 322 DAY 1 5 DAY 1 8 DAY 2 2 DAY 2 5 8. 075 7. 288 7. 054 7. 288 7. 558 7. 311 7. 315 6. 956 ECCLES Dispersalloy 9. 966 9. 620 10. 851 10. 590 11. 260 9. 070 9. 280 9. 014 7. 322 7. 922 9. 913 9. 279 8. 639 6. 809 7. 542 8. 672 9. 206 8. 685 8. 599 8. 480 7. 783 8. 270 7. 936 8. 997 FISCHER Valiant 5. 958 5. 829 4. 408 4. 533 4. 266 4. 473 5. 136 4. 460 5. 280 4. 762 4. 492 4. 279 4. 801 4. 505 4. 300 4. 862 4. 596 4. 704 4. 929 4. 867 6. 147 5. 798 5. 936 5. 468 GRUBE Valiant 6. 841 6. 904 6. 788 5. 782 8. 158 7. 740 7. 893 8. 026 12. 458 11. 878 11. 771 12. 404 12. 146 10. 693 10. 484 10. 221 13. 911 13. 421 12. 618 11. 176 11. 669 13. 439 13. 208 13. 090 MESSERMAN Dispersalloy 11. 357 11. 238 11. 887 12. 086 15. 335 14. 712 14. 473 15. 859 17. 796 17. 484 16. 765 19. 584 19. 321 20. 716 20. 696 19. 995 15. 336 14. 602 14. 086 18. 625 17. 759 12. 389 16. 285 15. 580

DENTISTS BRAND DAY 1 DAY 4 DAY 8 DAY 1 1 RUBIN Tytin 14. 207 13. 175 12. 244 11. 835 11. 639 11. 568 14. 147 13. 240 21. 055 20. 484 19. 769 20. 150 22. 512 20. 912 18. 798 16. 579 DAY 1 5 DAY 1 8 DAY 2 2 DAY 2 5 9. 407 8. 281 8. 693 9. 731 9. 556 11. 781 9. 799 9. 219 SUKEL Tytin 9. 024 8. 662 8. 272 7. 521 8. 043 11. 216 9. 515 8. 670 10. 757 10. 341 9. 713 6. 384 7. 030 7. 540 7. 428 6. 782 7. 539 7. 108 6. 656 6. 508 6. 899 6. 508 6. 959 8. 200 WILLIAMSO N Dispersalloy 11. 424 10. 897 12. 077 10. 392 10. 738 10. 976 12. 094 11. 538 8. 242 7. 675 8. 123 7. 425 7. 499 8. 872 8. 588 8. 463 10. 529 10. 427 10. 553 11. 149 10. 463 10. 156 10. 559 10. 228 YOO Tytin 9. 098 8. 063 7. 795 7. 366 7. 994 7. 304 9. 803 9. 305 10. 949 10. 216 10. 773 10. 431 12. 250 11. 319 12. 476 11. 197 15. 925 15. 525 14. 992 12. 234 12. 797 14. 670 14. 038 13. 647

? De. Rouen et al. JAMA 295, 1784 -92, 2006

ELEVATED MERCURY IN IDIOPATHIC DILATED CARDIOMYOPATHY (IDCM). WHERE DOES THE Hg COME FROM? LEVELS ng/g Hg Controls 8. 0 IDCM 178, 400 Sb 1. 5 19. 260 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. WHY HASN’T NIH REQUESTED PROPOSALS FOR RESEARCH TO STUDY THIS? ? THIS IS PROOF THAT MERCURY CAN CONCENTRATE IN SPECIFIC TISSUES OR ORGANS OF THE BODY, EVEN IF Hg BLOOD LEVELS ARE FOUND TO BE IN THE NORMAL RANGE.

• The previous slide shows that prolonged exposure to mercury vapor decreases the child’s ability to excrete mercury through their kidneys. • This is consistent with the well known toxic effects of mercury on kidneys. • This explanation is consistent with the reports by the EPA and National Academy of Sciences that 8 to 10% of American women have such high Hg body levels that would render increased susceptibility to neurological damage any child they would give birth to.

Current Situation • Drugs use to treat behavioral disorders (autism, ADHD) has increased dramatically (369%) in the recent past to where we now spend more on these drugs than we do on antibiotics and asthma drugs for children. • The USA now spends more on drugs to treat ADHD than it does on antibiotics and asthma drugs. • 1 of 6 children in the USA have a diagnosed neurodevelopmental problem according to the CDC. About 1 in 166 have autism. • We have a major problem! • Yet, our FDA, CDC, NIH and AMA ignore the effects of dental and medical induced exposures to mercury.

Axonal Transport - A Process Essential for the Survival of Neurons Dendrite Axon Membrane Bound Organelle Dynien Microtubule Kinesin

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

Hg. EDTA Induces Aberrant [32 P]8 N 3 GTP-ß -Tubulin Interactions Indicative of AD Alzheimer’s Disease Brain Normal Brain without and with Hg 2+.

Pink Disease/Acrodynia • Affected 1 in 500 children in late 1800 s until about 1940. • Cause was mercurous chloride (calomel) in teething powers. • Elimination of these mercury containing teething powders eliminated this disease. • Therefore, it is very plausible that low level exposure to mercury at an early age could cause a neurological disease. • Note: Ethylmercury is many times more toxic than mercurous chloride. Lethality is not an absolute measurement of all toxic effects.

Thimerosal Is Composed of Thiosalicylic Acid And Ethyl Mercury, A Known Neurotoxicant 1. The Merck Index, 12 th ed. , p. 1590, #9451 (1996). 2. Martindale The Extra Pharmacopoeia, 30 th ed. , 804 (1993).

Organ Mercury Levels in Infants with Omophaloceles Treated with Thimerosal. Fagan et al. Archives of Disease in Childhood 52, 962 -64, 1977 • Between 1969 -75, 13 cases were treated, 10 died. Mercury analysis of organs ranged from 65 to 2, 700 times normal levels. This appears to be from 9 to 48 topical applications of 0. 1% thimerosal applications. NOTE; These children were most likely on antibiotics. Consider the effect on their immune system! • “Paradoxically, (in another study) 3 infants exposed postnatally (Iraq, Methyl-Hg by ingestion) did not exhibit signs or symptoms, though their blood levels were >1, 000 ppb, and one was >1, 500 ppb. ” No antibiotics involved! Blood levels are not a measure of toxicity. • CONCLUSION IN 1977: “Organic mercurial antiseptics should be heavily restricted or withdrawn from hospital use, and the fact that mercury readily penetrates intact membranes and is highly toxic seems to have been forgotten. ” • Result: Mercurochrome and Merthiolate was removed from the

THE BIG MISTAKE! • YET SOME INDIVIDUALS AT THE CDC AND FDA DECIDED IT WAS OK TO INJECT THIMEROSAL INTO A NEWBORN INFANT AT LEVELS THAT WOULD BE EPA SAFE IF THE INFANT WEIGHED 275 POUNDS! • The EPA “safe level” was based on mercury exposure from eating fish and whale meat. • Most of the heavy metal protection in humans is in the intestinal area as we evolved eating and drinking contaminated food and water. This is bypassed on injection of thimerosal or breathing mercury vapor.

• • • • • • • • Table 1. US Department of Education statistics on autism in children aged 6 -21 served by Individuals With Disabilities Education Act (IDEA) State Alabama Alaska Arizona Arkansas California Colorado Connecticut Delaware District of Columbia Florida Georgia Hawaii Idaho Illinois Indiana Iowa Kansas Kentucky Louisiana Maine Maryland Massachusetts Michigan Minnesota Mississippi Missouri 1992 -1993 68 8 199 30 1, 605 14 164 15 0 582 262 52 39 5 273 67 74 38 409 37 28 493 288 296 0 336 2001 -2002 904 223 1, 348 774 13, 257 538 1, 470 294 144 4, 328 2, 462 380 356 3, 802 3, 262 554 743 1, 022 1, 297 552 2, 396 2, 681 4, 719 3, 270 461 1, 953 % Increase 1, 229 2, 687 577 2, 480 726 3, 743 796 1, 860 644 840 631 813 75, 940 1, 095 727 904 2, 589 217 1, 392 8, 457 444 1, 538 1, 005 481

Thimerosal is toxic to tubulin and actin. Combinations of Hg 2+ and thimerosal would be at least additive.

Observation • Thimerosal, or ethyl-mercury, is a potent and rapid inhibitor of many enzymes necessary for human health. • Thimerosal or ethyl-mercury does not have to break down to Hg 2+ to be toxic to these enzymes or structural proteins. • The inhibition of tubuline polymerization would disrupt neuronal connections and prevent the mitotic spindle formation needed for immune cell division. The latter would induce an immune system suppression.

MOST VACCINES CONTAIN TRACES OF THIMEROSAL EVEN IF IT IS NOT ADDED AS A PRESERVATIVE. The vaccine thimerosal concentration was (is) 125, 000 to 250, 000 nanomolar!

INCREASED NEURON DEATH DR. MARK LOVELL’S LAB

SYNERGISTIC EFFECTS OF HEAVY METALS IS QUITE COMPLEX AND CAN GREATLY ENHANCE THE TOXICTY OF MERCURY Shubert et al. Combined Effects in Toxicology--A Rapid systematic Testing Procedure: Cadmium, Mercury & Lead. J. of Toxicology & Environmental Health 4: 763, 1978. 1. “the administration of an essentially no response level (LD 1) of a mercury salt together with a 1/20 of the LD 1 of a lead salt killed all of the animals. ” 2. “Generally, a combination was synergistic when the most toxic member was present at or near its LD 1 dose in the presence of a much less toxic member. ” 2. Conclusion: Mixing borderline toxic levels of two toxic metals (Pb 2+ & Hg 2+) makes an extremely toxic solution.

SYNERGISTIC TOXICITIES Al: NEOMYCIN: TESTOSTERONE EFFECTS 50 NANOMOLAR THIMEROSAL DR. MARK LOVELL COLLABORATOR + TESTOSTERONE

Recent Publication On Thimerosal Exposure and Neurological Disorders: Autism Effects of Thimerosal on Nerve Growth Factor Signal Transduction and Cell Death in Neuroblastoma Cells. Parran et al. , Toxicological Sciences, 2005. Data demonstrated that thimerosal could alter NGFinduced signaling at concentrations lower than those causing neuronal death. Therefore, the neurons growth and properties could be impeded at exceptionally low levels of thimerosal without killing the neurons.

Estradiol Reduces Cumulative Mercury and Associated Disturbances in the Hypothalamus-Pituitary Axis of Ovariectomized Rats. Oliveria et al. Ecotoxicol. Environ. Safety Jan. 10, 2006 • Methyl-mercury induced a decrease in LHRH in the medial hypothalmus and a decrease in plasma levels of LH. These decreases in LHRH and LH were abolished by estrogenic replacement therapy. • “The estrogenic effects were associated with a reduction of mercury content of the anterior pituitary gland medial hypothalmus, suggesting a protective estrogenic effect. ”

Observations 1. The toxicity of Hg 2+ and thimerosal is dramatically enhanced by other heavy metals, antibiotics and testosterone. 2. It appears as if infants are much more susceptible to mercury toxicity than more mature individuals. 3. Female hormone prevents mercury retention and damage in certain areas of the brain.

Mercury Effects on the Immune System • The mitotic spindle is built on tubulin quite similar to that found in axons of neurons. Therefore, since the cells of the immune system must divide for an effective immune response Hg inhibits this and actively suppresses the immune system. • Thimerosal is a very potent inhibitor of phagocytosis by mononuclear phagocytes, inhibiting the process at low nanomolar levels. (Rampersad et al. , Transfusion 45(3): 384 -93, 2005). This prevents removal of microbes and ethyl-Hg damaged cells and proteins leading to greater susceptibility for microbe infection and widespread autoimmune problems.

INNATE VERSUS ADAPTIVE IMMUNITY • Innate immunity is the immunity infants are born with and refers to antigen-nonspecific defense used to kill micobes by ingestion and digestion by white cells (neutrophils and macrophages), the first step is PHAGOCYTOSIS, which is shut down by levels of thimerosal reported to be 1 nm. • After phagocytosis the dissembled microbe parts are released by the macrophage to the B-cells of the immune system which presents them to the T-cells for production of antibodies to the microbe antigenic sites. This is the adaptive immunity part, where specific antibodies are made against the specific microbe. • Bottom line, a newborn infant freshly injected with thimerosal cannot take advantage of innate immunity due to blockage of phagocytosis and consequently will have impaired adaptive immunity for some time period.

Epidemiological Studies • A study on seven-year-old children in the Faeroe Islands found that blood pressure problems increased with decreased blood Hg. This implies retention toxicity effects of Hg in this comparison. • In the Sechylles study of >700 children, boys with higher levels of hair mercury performed better on some tests as the Boston Naming test. This implies that ability to excrete increases hair Hg levels, not exposure, in this comparison. • CONCLUSION: Blood and hair Hg levels are not a measure of exposure at low levels, but rather a measure of both exposure and ability to excrete mercury.

MERCURY BIRTH HAIR LEVELS VS. AMALGAM FILLINGS IN AUTISTIC AND CONTROL GROUPS Hair Hg level (mcg/g) Number of amalgams: Control: autistic ratio: N: Data from A. Holmes, M. Blaxill & B. Haley, Int. J. of Toxicology v 22, 2003 Controls Autistic 0 -3 2. 64 15 4 -5 6. 93 22 6 -7 6. 70 29 8 -9 6. 32 30 >10 17. 91 43

BIRTH-HAIR MERCURY OF AUTISTIC VS. CONTROL GROUPS HIGH Female Male Hair Hg level (ppm) AMALGAM COUNT Data from Amy Holmes, Mark Blaxill & Boyd Haley, Int. J. Toxicology v 22, in press, 2003 LOW Non-autistic Mean=3. 79 n=34 Autistic Mean=0. 47 n=94

Birth-Hair Hg Levels In Infants From Mothers With More Than Eight Amalgams • Data (Hg ppm) was selected from only mothers who had 8 to 15 amalgam fillings to determine if these extremes showed a more definite trend on the differences between autistic and normal infants. The ratio was 12: 1.

BIRTH-HAIR MERCURY BY SEVERITY OF AUTISM Female Male Hair Hg level (ppm) Data from Amy Holmes, Mark Blaxill & Boyd Haley, Int. J. Tocicology v 22, in press, 2003. Mild Mean=0. 71 n=27 Moderate Mean=0. 46 n=43 Severe Mean=0. 21 n=24

Observation • Autistic infants in utero appear to have an impaired ability to excrete mercury when compared to normal children! • New concept: Low mercury levels in the hair and blood do not imply lack of toxic mercury exposure or retention in the body!

The involvement of the 2004 Institute of Medicine (IOM) report. • The 2004 IOM committee was funded by the CDC. • The 2004 IOM report cleared thimerosal as being involved in autism and recommended that no further research be done on this issue. • The 2004 IOM report was based only on 5 epidemiological studies of questionable value. • The 2004 IOM report totally dismissed the basic science research on thimerosal toxicity and the resultant aberrant biochemistry possibly caused by mercury-like toxicity reported by several research scientists. • Genetic studies were suggested yet the 2007 Autism Geneome Project could find no definitive genetic linkage to autism causation. • A recent congressionally requested NIH committee looked at the 2004 IOM report and, in my opinion, gave it a very bad evaluation.

Pre-IOM 2004 Science On Hg Exposure and Autism • Holmes et al. showing that autistic infants do not excrete mercury as effectively as do normal children. • Wakefield studies on increased measles virus in intestines of autistics. • Results from Dr. Bradstreet on increased Hg body burden of autistics. • Early clinical Results of Drs. Stepanie Cave & Amy Holmes using DMPS to successfully treat autism. • Results of Dr. Mady Horning on thimerosal inducing autism like symptoms in mice susceptible to autoimmune disease. • Results of Dr. Richard Deth with regards to methyl-B 12 production. • Results of Dr. Jill James on reduced GSH levels in autistics. • Epidemiological studies of Dr. Mark and David Geier. • The Institute of Medicine totally dismissed all of these biological studies plus others and did not present any results to counter the above studies, and based their opinion primarily on epidemiological studies done by those with a conflicting interest using invalid data bases. • Read “Evidence of Harm by David Kirby” and the Safe Minds website for Freedom of Information Act materials.

Who did the Epidemiological Studies the IOM depended on? ? • The Verstraten studies at first showed autism rates were enhanced by thimerosal exposure. All the CDC data was lost or destroyed after it was published. Verstraten now works for a major vaccine producer in Europe. • Two studies were done by Danish (Madsen and Hviid) who worked for the Stantens Serum Institute (SSI). SSI makes thimerosal containing vaccines and sells them to other countries because they are not allowed to be used in Denmark since 1992. • One study was done in England by E. Miller. After her results were made known to the IOM the National Health Service removed thimerosal from English vaccines. • Troubling, that the opinion of the FDA is based totally on foreign, conflicted opinions. Why couldn’t the CDC find epidemiologists in the USA to do these studies? ? ? • A recent call by the IOM to discuss possible environmental causes of autism in 2007 explicitly excludes vaccine considerations.

Autism Risks From 5 Sequential Studies by Verstraten et al. of CDC Study 1 7. 62 (1999) Study 2 Study 3 Study 4 Study 5 2. 48 1. 69 Simpsonwood Meeting 1. 52 (2001) 1. 00* (2005) *i. e. , no increased risk of autism compared to low exposure group After publication in 2005 all of the data for this work was “lost” by the CDC!!! Go to Safeminds. org to read the FOIA material on the Verstraten studies. Evidence of Harm

DANISH STUDY 1. In USA rate is 1/166 or 60/10, 000! 2. Outpatients added in 1995. 3. Large Copenhagen Clinic added in 1992. 4. Autism classification changed in 1994. 5. Thimerosal removed from vaccine. Conclusion; exposure to a potent neurotoxin, thimerosal, prevents autism!!! Nonsense!

Ties between epidemiology studies and vaccine manufacturers Study Vaccine Manufacturer Tie Verstraeten et al. 2003 Verstraeten employed by GSK starting 7/16/01 Study CDC funded Madsen et al. 2003 2 coauthors employed by the SSI Study CDC funded Hviid et al. 2003 All 4 coauthors employed by the SSI Study CDC funded Stehr-Green et al. 2003 Stellfeld (coauthor) was Head of the Department of Medicine, SSI Miller et al. 2004 Miller has served as an expert witness for GSK, Aventis Pasteur and Merck (all named as defendants in ongoing civil litigation) Study WHO funded through CDC

Dr. Mark and David Geier Studies

CONCLUSIONS • Denmark and Sweden did not appear to have an autism epidemic. They have always vaccinated later in life and used lower doses of mercury than the USA. Using their database to study the autism epidemic is like doing epidemiology on the effect of mosquitos on malaria and doing the study in Alaska instead of Panama. • For analysis of the CDC funded “Verstraten studies” visit the various autism websites that present the FOIA obtained documents that discuss the problems with this study. • A recent NIH committee evaluated the 2004 IOM report and called the epidemiological studies used to call thimerosal not involved as seriously flawed. • The NIH committee recommended that “new methodologies be used to evaluate thimerosal involvement.

• Other Considerations In England, between 1970 -1980, about 14. 7% of children were not vaccinated as suggested. Yet a parental autism group there report (Tony Bateson), on the internet, only two cases of autism in non-vaccinated children were found in their search of autistics born during this time frame. • The recent UPI series on autism by Dan Olmstead finds: 1. Very little, if any, autism in the Amish as reported by. 2. Healthfirst, a Chicago Clinic that does not vaccinate in the first year of birth reports no autistic children born since 1985 from a population of about 35, 000 children. • The dramatic increase in autism in China following the end of the cold war and the importation of Western vaccines (Evidence of Harm by David Kirby).

Conclusions 1. 2. 3. 4. It is my opinion that the CDC and FDA do not always have the interests of the USA citizens at the top of their list. Make legislation at the state level that is more difficult to control by big pharmaceutical companies. Note that the FDA has ruled that their approval over ruled any state level legislation! If you know a child with autism spectrum disorder inform the parents that autism is treatable. In many cases it is a form of early infant mercury toxicity. Dr. Robert Nataf’s laboratory reports that many children diagnosed as “autistic” have prophyrin profiles indicating mercury toxicity. This is based on the research of Dr. James Wood of the University of Washington. 5. Get a urinary porphyrin profile done on your child to determine the possibility of mercury toxicity. Send the results to A-CHAMP. org. The compilation of these results will be presented to legislators throughout the USA.

THE SMOKING GUN STUDY • Done in Paris, France (since the 2004 IOM committee recommended NIH not fund thimerosal studies) in a large autism clinic. • Investigated porphyrin profiles in autistic versus normal children because these profiles are the best indicator for heavy metal toxicity, especially mercury toxicity. • Found porphyrin profiles that indicated 53% of autistic children surveyed were mercury toxic. • Reversed toxic porphyrin profiles by treating autistics with a mercury chelator. Therefore, the cause was not genetic, but mercury toxicity. • Supporting data from Norway has been reported. • Dr. Robert Natal and Dr. Richard Lathe were the lead researchers in this work published in the International J. Toxicology 2006.

PORPHYRIN BIOCHEMISTRY RATES OF EACH ENZYME RX IS DIFFERENTIALLY AFFECTED BY Hg 2+ VERSUS OTHER

WHAT ARE PORPHYRINS? • Porphyrins are a class of compounds that lead to the synthesis of heme, the iron binding red compound of hemoglobin that binds oxygen and aids in delivery to cells, where it is used in the mitochondria to help make energy (ATP). Lack of heme or hemoglobin leads to a very pale complexion (ever notice the complexion of autistic children? ) • Heme has other biological uses. It is in the electron transport pathway that makes ATP. A shortage of heme would prevent adequate energy production. • Heme is needed for active P 450 enzymes, the enzymes that modify organic toxins and aid in removing them from the body. • Heme is needed to remove amyloid protein from human brain to prevent production of amyloid or senile plaques as identified with Alzheimer’s diseased brain.

DATA FROM THE USA • I HAVE RECEEIVED OVER 20 URINARY PORPHYRIN PROFILE RESULTS OF AUTISTIC CHILDREN IN THE USA. ALL INDICATED MERCURY TOXICITY WITH VALUES IN THE ABOVE 500 NANOMOLES/GRAM CREATININE. • THE REFERENCE VALUE IS 50 TO 90.

A “CROSSOVER POINT”

Effects of Antibiotics, Diet and other Metals on Hg Excretion: Found in Published Literature • Rats exposed to antibiotics were severely impaired in their ability to excrete mercury. • Rats on milk versus high protein diets were much less able to excrete mercury. • The great enhancement of synergistic toxicity with Hg and other heavy metals (e. g. lead) is well documented in the literature. We have many children with other heavy metals in their bodies. • The above confounders have rarely been considered by those who write articles supporting the safety of thimerosal or dental amalgams.

Hg & THIMEROSAL DISPLAY ADDITIVE TOXICITIES.

RAPID BLOOD TO BRAIN MOVEMENT OF [203 Hg]THIMEROSAL: Gasset et al. Tetratogenicities of Opthalmic Drugs. Arch. Opthalomology 93, 52 -55, 1975. • Pregnant rabbits were injected subcutaneous with [203 Hg]thimerosal. • From hour 1 post injection to hour 6 the cpm of 203 Hg in the blood decreased from 100, 000 to less than 25, 000 cpm, or over 75%. • From hour 2 post injection to hour 6 there was increased cpm of 203 Hg in the fetal brain (2 fold), liver (4 fold) and kidney (3 fold). • Yet the IOM/CDC/AAP states that the rapid loss of mercury from thimerosal from the blood makes it unlikely to be toxic enough to cause autism. Pichichero et al. Lancet 360: 1737, 2002

IATROGENIC EXPOSURE TO MERCURY AFTER HEPATITIS B VACCINATION: Stajich et al. J. Pediatrics 136; 5; 679, 2000, • • • Vaccine Hg = 0. 5 ml of 125, 000 nanomolar, or 12. 5 mcg Hg. Normal babies were about 3. 5 kg or about 7 lbs with 0. 04 mcg Hg/liter blood. At 16% blood volume estimate then 0. 560 kg was blood or 0. 560 liters. (125, 000 nmoles/liter)(0. 0005 liter) = (0. 56 liter)(infant blood Hg nmoles/liter) Calculated infant blood Hg level = 112 nanomoles/liter if all Hg was to be in the blood. • Blood Hg levels were determined pre (0. 04 mcg/liter)and 48 to 72 hours post injection (2. 24 mcg/liter). • 2. 24 mcg/liter divided by 200 g/mole for Hg = 11. 2 nanomoles/liter of Hg in “actual testing”, or 10 fold less was found in the blood. Therefore, they concluded that blood levels were to low to be toxic! • 11. 2 nanomoles/liter is about 10% of 112 nanomoles/liter, where did 90% of the Hg go? (Of 62. 5 nanomoles injected, 6. 27 nanomoles were found in the blood. ) • This 11. 2 mcg (90%) of mercury that is not accounted for after 2 -3 days and is either somewhere else in the body or has been excreted. Which is it? REQUIRES A MATERIALS BALANCE SHEET TO DETERMINE THIS.

Infant Hg Excretion Rates Based on Pichichero et al. Lancet 360: 1737, 2002 Data and a 4 kg Infant Stool Hg concentration Daily Hg excretion Days to excrete X mcg 12. 5 mcg 187. 5 mcg (ng/g) (mcg/day) (days) Minimum: 23 0. 09 -0. 28 44. 6 -139 670 -2, 083 Mean: 82 0. 33 -0. 98 12. 7 -37. 9 191 -568 Maximum: 140 0. 56 -1. 68 7. 44 -22. 3 112 -335 As previously calculated, 11. 4 mcg of Hg were not accounted for after a single vaccination and had to be excreted or retained by body tissues within 48 -72 hours, or 2 -3 days. As can be seen in the table above, it takes as a minimum about 7. 4 days to excrete 12. 5 mcg Hg using the maximum fecal Hg excretion rate. This includes an assumption of 1 -3 g feces/day/kg body weight. Using the mean max daily excretion (0. 98) then 1. 96 -2. 94 mcg of Hg of the 12. 5 mcg injected would be excreted leaving 10. 54 -9. 56 mcg or 84 to 76% Hg unaccounted for at days 2 -3 respectively.

Results of Hg Materials Balance • Initially injected thimerosal rapidly leaves the blood and goes into the cells of the body and CNS. • Measuring the Hg levels in the blood hours or days after thimerosal exposure cannot be used to determine the extent of mercury exposure or the amount retained in the body. • Fecal excretion of thimerosal delivered mercury is to slow to account for the drop in blood Hg levels. Also, this drop does not represent removal from the body, even for normal children. Autistic children would be even less able to excrete Hg via the feces. • Even normal children had a 6 -fold difference in excretion rates according to Pichichero et al. • How much lower would the autistic children be in rates of Hg excretion?

RESULT • The often used argument by pro-thimerosal advocates that the ethylmercury clears the blood to quickly to be toxic displays a lack of complete and intelligent analysis of the data. Don’t accept this argument! • Early blood mercury loss is most likely a measure of partitioning of Hg into the body and not excretion.

BODY RESULTS OF ETHYL-Hg EXPOSUE • INFANTS INJECTED WITH THIMEROSAL AND UNABLE TO EXCRETE IT MAY ARE THE EQUIVALENT OF A BIOCHEMICAL TRAINWRECK----AND ALL TRAINWRECKS ARE NOT ALIKE. • NOTE THE SIMILARITY OF AUSTISM SYMPTOMS TO ORGANIC-MERCURY POISONING AS SEEN IN THE MINAMATA BAY DISASTER. CONSIDER AGE OF EXPOSURE—INFANTS RESPOND DIFFERENTLY TO Hg THAN ADULTS.

Deposition of Body Mercury into the Kidney • Chelators that bind mercury weakly may increase the mercury levels in the kidney by removing it from the blood and other tissues and depositing it in the kidney. This was the explanation for the increased kidney toxicity on treating mice with DMPS and Nacetylcysteine after exposing them to mercury by injection. Brandao, R. et al. “DMPS and Nacetylcysteine Induced Renal Toxicity in Mice Exposed to Mercury. ” Biometals (2006) 19: 389 -398

WHY DMSA/DMPS ARE NOT REAL CHELATORS EVEN THOUGH THEY DO REMOVE MERCURY. /-S-Hg-S-/ Hg 2+ DMSA & DMPS WERE DEVELOPED IN THE 1940 s! x MUST BE LINEAR AND IT CANNOT BE WITH –SH GROUPS ON ADJACENT CARBONS. WHAT FORMS IS THE EQUIVALENT OF DMSA-S-Hg-S-DMSA OR DMSA-Hg-CYSTEINE TYPE LINKAGES.

New Chelator Concept • Most available chelators are water soluble because they carry ionic charges. This includes DMPS, DMSA, glutathione, and Se 2 -. Therefore, they are not efficient at removing Hg 2+ that is located in fatty (hydrophobic) environments in cells. Additionally, water soluble chelators are rapidly excreted in the urine. • Most mercury in the body is not available to DMPS, etc. for binding as they are intracellular or in hydrophobic locations. • A new chelator releases a very effective mercury chelator that enters hydrophobic areas where most mercury may be located. Because of its hydrophobic nature it stays in the body for a much longer time. • P-450 enzymes may help in the excretion of this type of chelator with mercury bound.

First Generation Mercury Chelating Agents Base compound binds Hg 2+ tighter and more selectively than anything known to date. R-S-Hg-S-R linear Benzene bis-amido bis-thiol Pyridine bis-amido bis-thiol Water insoluble, but lipid soluble

Glutathione derivative of Functionalized Mercury Chelating Agents Making them Water Soluble Note: Molecule would be charged and water soluble at p. H 7. 4. Glutathione Very water soluble Glutathione

Toxicity Study of Chelator • • • Group Test 1 Test 2 Test 3 Total A 0 0 • Test 4 0 B 100 200 300 600 - C D 200 300 400 500 900 1, 200 1, 500 • Procedure: Rats were injected under the skin in the stomach area with chelator to the amount in μMoles/kg body weight. Three days pause was between each treatment. • Result: No toxicity or weight loss was observed.

Protection Against Mercury Toxicity • Rats given a lethal dose of Hg 2+ died within 3 days exhibiting head tremors and convulsions before death. • Rats similarly injected with Hg 2+ but 20 minutes later given the new chelator did not show any signs of toxicity except to drink more water and appear slightly less energetic for two days. They fully recovered and are now quite active after 3 months. No signs of tumors, etc. yet. • Last month, a commercial toxicology laboratory has confirmed that the new chelator is not toxic at 5 grams/kg body weight, the highest testing level! Two thirds of the way through a 28 day trial giving 1. 0 g/kg body weight has yet to uncover any toxic effects on mice. • This research is being done for the purpose of obtaining FDA approval for these new chelators.

Acknowledgements • Mr. H. B. Wallace, Scottsdale, AZ (deceased) • Mr. John and Carol James, Hertfordshire, GB