Transcript Italian Talk - Life Sciences Research Organization

MERCURY TOXICITY: Dependence on
Dose, Diet and Genetic Susceptibility
DR. BOYD E. HALEY
PROFESSOR AND CHAIR
DEPARTMENT OF CHEMISTRY
UNIVERSITY OF KENTUCKY
VISUALIZATION OF MERCURY
EMITTING FROM A DENTAL AMALGAM


From: www.
uninformed
consent.com
David
Kennedy’s
IAOMT tape
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 H2O.
H2O was changed daily and Hg level measured by a
mercury cold vapor analyzer.
 Without brushing the amalgams released 7.54g
Hg/cm2/day.
 With two 30 second brushings with a medium
toothbrush the amalgams released 45.5g
Hg/cm2/day, a six-fold increase.
 This confirms the earlier observation of Dr. Stock in
1926 in Zeitschr. Angew. Chem. 39, 461-466.

Neuron Survival (% Initial Number)
120
Hg & THIMEROSAL DISPLAY ADDITIVE TOXICITIES
TO NEURONS IN CULTURE.
100
80
Control
50 nM thimerosal
60
50 nM thimerosal
+10 nM HgCl2
50 nM thimerosal
+ 25 nM HgCl2
40
10 nM HgCl2
25 nM HgCl2
20
0
5
10
15
20
Time (hr) After Treatment
25
30
MERCURY BIRTH HAIR LEVELS VS.
AMALGAM FILLINGS IN AUTISTIC AND
CONTROL GROUPS
14
Control
12
10
Hair Hg level
(mcg/g)
8
Data from A. Holmes,
M. Blaxill & B. Haley,
Int. J. of Toxicology
v22, in press, 2003
6
4
2
0
Number of amalgams:
Control: autistic ratio:
N:
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
ACTUAL VERSUS PREDICTED BIRTH HAIR
MERCURY LEVELS
Hair Hg level= (5.60)+0.04(amalgam volume)+1.15(fish consumption)+0.03(vaccine):
R2= 0.79
20
Predicted hair
Hg level
(mcg/g)
15
Controls
Autistics
10
5
0
-5
0
Data from A. Holmes,
M. Blaxill & B. Haley,
Int. J. Toxicology, in
press, 2003
5
10
15
Actual hair Hg level (ng/g)
20
BIRTH-HAIR MERCURY BY
SEVERITY OF AUTISM
1.4
Female
Male
1.2
Hair Hg
level
(ppm)
Data from Amy
Holmes, Mark
Blaxill & Boyd
Haley, Int. J.
Tocicology v22, in
press, 2003.
1
0.8
0.6
0.4
0.2
0
Mild
Mean=0.71
n=27
Moderate
Mean=0.46
n=43
Severe
Mean=0.21
n=24
CONTRAST BETWEEN BIRH HAIR Hg
LEVELS AND BODY Hg LEVELS
 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.
 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.
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.
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.
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?
ELEVATED MERCURY IN IDIOPATHIC
DILATED CARDIOMYOPATHY (IDCM).
WHERE DOES Hg COME FROM?
LEVELS ng/g
Hg
Sb
Controls
8.0
1.5
IDCM
178,400
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.
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.”
 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.

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.


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.
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).



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!
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.
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.
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).”
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.
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”
“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.”
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.
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.”
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.
 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.”

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.
 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.

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.
SYNERGISTIC TOXICITIES on NEURONS IN CULTURE
Neuron Survival (% Initial Number)
120
Control
Al:NEOMYCIN:TESTERONE
EFFECTS
100
50 nM thimerosal
500 nM Al(OH)3
1.75 µg Neomycin/ml
50 nM Thimerosal
500 nM Al(OH)3
80
50 nM Thimerosal
1.75 µg Neomycin/ml
50 NANOMOLAR
60
50 nM Thimerosal
500 nM Al(OH)3
1.75 µg Neomycin/ml
40
DR. MARK
LOVELL
20
COLLABORATOR
+ TESTOSTERONE
0
0
5
10
15
20
Time (hr) After Treatment
25
30


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.
Axonal Transport - A Process
Essential for the Survival of Neurons
Dendrite
Axon
Membrane
Bound
Organelle
Dynien
Kinesin
Microtubule,
made of
polymerized
tubulin dimers.
Autoradiogram of [32P]8N3GTP Photolabeled
Control & AD Brain Hippocampus Homogenates
EDTA Prevents Cd, Cu & Zn, But Not Hg,
Inhibition of [32P]8N3GTP Photolabeling of
Brain ß-Tubulin
Partitioning of ß-Tubulin is Aberrant
in Alzheimer’s Diseased Brain
Homogenates
NORMALS
NORMALS
NORMALS
ALZHEIMERS
ANTIBODY DETECTION OF BETA-TUBULIN
MERCURY AND ALZHEIMER’S DISEASE

Exposure of neuroblastoma cells to 10-9 molar mercury
increases Tau phosphorylation and secretion of betaamyloid. 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
Therefore, Hg exposure can create these diagnostic
hallmarks of Alzheimer’s Disease!
HgEDTA Induces Aberrant [32P]8N3GTPß-Tubulin Interactions Indicative of AD
EFFECT OF SEQUENTIAL AMALGAM EXTRACTION
SOLUTIONS ON THE VIABILITY OF BRAIN TUBULIN
120
100
80
60
40
20
Hours of Amalgam Soak
to
12
0
96
96
to
72
72
to
48
48
to
24
24
to
12
to
12
8
8
4
to
4
2
to
2
1
to
1
to
0
nt
ro
l
0
co
%
Active
PROTEINS INVOLVED IN Hg EXCRETION




GLUTATHIONE (carries Hg through bilary transport,
rapidly oxidized by excess Hg2+. Note: Hg2+ 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.
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.
Biochemical Differences Between the Three Most
Common Apolipoprotein E Isoforms is Reflected
in Hg Binding Capacity
2 Mercury binding
sites
LOW RISK FOR EARLY AD
1 Mercury binding
site
No Mercury binding
sites
HIGH RISK FOR EARLY AD
RELATIONSHIP TO NUMBER OF APO-E –
SH GROUPS AND AGE OF AD ONSET
APO-E

2
2
3
4
4 (>90)
3 (>90)
2 (80-90)
3
3 (>90)
2 (80-90)
1 (70-80)
4
2 (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
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.
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.
The 3/3 homozygote was found at 54.8 and 51.4 in the
symptomatic and blood donor groups, respectively.
Effect of Removal of Amalgams & Antioxidant Therapy on
Patient Symptoms and Blood Hg Levels.
Controls matched with test groups in age, gender & socio-economic conditions. They
were subjectively healthy persons not suffering from any disease.
U. Lindh et al., Neuroendocrinology Letters, v23, 459, 2002.
CONCLUSIONS WITH REGARD TO AD




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 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!
CONCLUSION REGARDING
AMALGAM FILLINGS


DUE TO THE ENHANCEMENT OF Hg TOXICITY
AND RETENTION BY FACTORS FOUND IN THE
DIET, ANTIBIOTICS, OTHER TOXICANTS SUCH
AS Pb2+ & Cd2+, 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.