Monday, March 26, 2007

Abstract of low NO hypothesis of ASDs

Nitric oxide is important in many physiological processes which are known to be disrupted in autism. Regulation of neural proliferation requires NO. The formation and refinement of neural connections requires NO. Low NO decreases the number and extent of neuronal connectivity. The sensitivity of a neural network to change depends exponentially on connectivity in the near percolation region. Increased sensitivity to seizures, savant-type abilities, and deranged attachment are consistent with decreased connectivity and increased closeness to the percolation threshold in the CNS. Gut disturbances, obstructive sleep apnea, enuresis, decreased heart rate variability are consistent with decreased connectivity and increased closeness to the percolation threshold in the ANS. Below the percolation threshold, the functionality of a neural network would collapse.

A biofilm of autotrophic ammonia oxidizing bacteria in vivo oxidizes ammonia from sweat into nitrite and produces NO, some of which is absorbed into the skin. Modern bathing practices will remove a biofilm of autotrophic ammonia oxidizing bacteria and prevent its reestablishment. All NO mediated pathways are sensitive to the background NO level. A perturbation to the background level of NO will affect all NO mediated pathways with no threshold.

In the “wild”, before modern bathing practices, it would be impossible for humans to not develop a biofilm of these bacteria which are universally present in all soils and all natural water sources. Loss of those bacteria will affect the background NO/nitrite level, which will perturb physiology with no threshold. Low NO from the loss of these bacteria will perturb physiology in the direction of low NO. Normally, low NO is a “stress” state, a state invoked in response to “stress”. Adaptable in the short term, when metabolic resources need to be mobilized to deal with the cause of the “stress”, bad in the long term because “low priority” stuff gets put off. How much will physiology be affected? A good question, the answer of which will require an understanding of the thousands of coupled non-linear pathways regulated by NO, many of which are no doubt idiosyncratic.

Application of autotrophic ammonia oxidizing bacteria is a natural and non-invasive method for increasing the NO level in humans that is under normal physiological regulation via sweating. Increasing the background NO level in ASD children and adults should affect all NO mediated pathways in a normalizing direction and may improve or ameliorate some autism symptomology.

Introduction to the low NO cause of ASDs

Individuals with Autism Spectrum Disorders (ASDIs) are characterized by difficulties in communication, social interaction and by repetitive behaviors. The cause(s) of ASDs remains unknown. The complex genetic disorder hypothesis posits ASDs are emergent disorders of multiple genes. In ASDs, there is a high (but not absolute) concordance between monozygous twins,[i] moderate concordance between dizygous twins,[ii] [iii] and lesser concordance between siblings. The clustering of ASDs in families, suggests genetics, but the lack of complete concordance in monozygous twins shows environmental factors are important too. The increased concordance in dizygous twins over full siblings shows that the in utero environment is important. The incidence of monoamniotic twins is quite low, perhaps insufficient for statistical information on autism rates, but monoamniotic twins can be discordant for anencephaly[iv] implying that a shared amniotic sack or placenta is not necessary or sufficient to prevent discordance in major neurological development disorders. With no generally accepted environmental cause, ASDs are thought to be primarily genetic in origin with associations of perhaps 135 genes[v]. No doubt the complex effects on brain structure and behavior observed in ASDs are not mediated via a single pathway, but calls to abandon a search for a single explanation are premature[vi]. Nitric oxide is a pleiotropic signaling molecule used in thousands of metabolic pathways where it regulates, ATP supply, O2 consumption, steroid physiology, transcription, axon targeting, epigenetic programming and many other aspects of physiology and neurodevelopment. Stress is a low NO state.[vii] I suggest that low NO in utero, brought about by maternal stress leads to the ASD phenotype in affected individuals, and the genotype that leads to the ASD phenotype was adaptive under conditions where humans evolved, in the “wild”, but is perhaps now non-adaptive due to environmental change(s). What possible advantages could the ASD phenotype hold? The most characteristic feature of ASDs is a larger brain[viii], with smaller and more numerous minicolumns.[ix]

I suggest that the diagnostic criteria for autism are the dysfunctional extreme end of the characteristic spectrum that makes humans tool making and tool using. People with less extreme ASD symptoms have Asperger’s, or can pass for “normal”, or can even be “normal” or neurologically typical individuals (NTIs). I suggest that the division between individuals who are “autistic” and “non-autistic”, is just as arbitrary as a division between individuals who are “short” and “tall”. Just as arbitrary, but vastly more complex. Being too short, or too tall, are not considered disorders, however increasing height through pharmacological (dietary supplements, growth hormone), or sartorial (high heels or lifts) mechanisms, is commonly practiced because of the adverse effect that short stature has on success in relationships with other humans. ASDs affect relationships with other humans also, and many parents of ASDIs would do “anything”, to “fix” the ASD in their child.

The ASD characteristics: difficulties in communication, social interaction and repetitive behaviors, I hypothesize are features essential during the evolution of tool making and tool using behaviors and abilities. The repetitive behavior is necessary to develop the de novo skill to manufacture stone tools. Disrupted communication is necessary to achieve sufficient social isolation, necessary for ASD individuals to ignore the “conventional wisdom” (which may be wrong) as practiced by other members of his clan or tribe. This is the essence of what Thomas Kuhn spoke of in the Structure of Scientific Revolutions[x]. To make new paradigms, one must be able to work beyond and outside of the old paradigms. This is something that “normal science” does very poorly. Peer review is good for reviewing “normal” science; it is terrible for paradigm breaking science. “Conventional wisdom” is good if the environment has remained static since the “conventional wisdom” was first adopted. Obviously a changing environment or changes in human behavior can obsolete “conventional wisdom” requiring change, otherwise we would still be living in caves, wearing skins and using stone tools, or perhaps still living in trees, or in primordial ooze. We will never know when the first ancestor(s) of humans abandoned the “conventional wisdom” of his/her group and did something new which was successful. That early human ancestor(s) did, and by doing so flourished, is beyond doubt.

[i] Kates WR, Burnette CP, Eliez S, Strunge LA, Kaplan D, Landa R, Reiss AL, Pearlson GD. Neuroanatomic Variation in Monozygotic Twin Pairs Discordant for the Narrow Phenotype for Autism. Am J Psychiatry. 2004 Mar;161(3):539-46.

[ii] Greenberg DA, Hodge SE, Sowinski J, Nicoll D. Excess of twins among affected sibling pairs with autism: implications for the etiology of autism. Am J Hum Genet. 2001 Nov;69(5):1062-7.

[iii] Visscher PM. Increased Rate of Twins among Affected Sib Pairs. Am J Hum Genet. 2002 Oct;71(4):995-6; author reply 996-9.

[iv] Lim KI, Dy C, Pugash D, Williams KP. Monoamniotic twins discordant for anencephaly managed conservatively with good outcomes: two case reports and a review of the literature. Ultrasound Obstet Gynecol. 2005 Aug;26(2):188-93.

[v] Herbert MR, Russo JP, Yang S, Roohi J, Blaxill M, Kahler SG, Cremer L, Hatchwell E. Autism and environmental genomics. Neurotoxicology 2006 Sep;27(5):671-84.

[vi] Happe F, Ronald A, Plomin R. Time to give up on a single explanation for autism. Nat Neurosci. 2006 Oct;9(10):1218-20.

[vii] Esch T, Stefano GB, Fricchione GL, Benson H. Stress-related diseases – a potential role for nitric oxide. Med Sci Monit. 2002 Jun;8(6):RA103-18.

[viii] Herbert MR. Large Brains in Autism: The Challenge of Pervasive Abnormality. Neuroscientist. 2005 Oct;11(5):417-40.

[ix] Casanova MF, Buxhoeveden DP, Switala AE, Roy E. Minicolumnar pathology in autism. Neurology. 2002 Feb 12;58(3):428-32.

[x] Thomas S. Kuhn. The Structure of Scientific Revolutions. 3d edition. University of Chicago Press, 1996.

Discussion of the false "mercury causes autism" idea

I include this section solely because of the difficulty I have had in getting anyone to pay attention to my low NO hypothesis sufficiently to understand it, while paying great attention to ideas which are obviously and demonstrably wrong. I am particularly annoyed with the “mercury causes autism” idea, which is completely wrong, and for which there is no physiological explanation, or any actual data supporting it and which contradicts much of which is well known about ASDs and mercury physiology. I call it an “idea”, because it does not meet minimal standards for “hypothesis”. It is not consistent with much that is well known. A “hypothesis” that is inconsistent with much that is well known is rejected (by scientists) as a failed hypothesis (as the “mercury causes autism” idea should be). People with ASDs do not have the most important and diagnostic symptom of mercury poisoning, an elevated level of mercury. We are now some 6 years beyond the suggestion that autism is caused by exposure to mercury[i], and there has not been a single publication demonstrating any association between elevated mercury exposure, or elevated mercury levels and autism or severity of ASD symptoms. Mercury is easy to measure, cheaply and with high precision in minimally invasive specimens, blood, urine, hair. There have been multiple industrial human exposures to mercury, which have been accurately measured and toxicity symptoms correlated with accurately measured levels in hundreds or thousands of individuals. There have been publications showing no association between mercury and autism.[ii] The proponents of the “mercury causes autism” idea have continued to press their case in the popular press, and in the courts, and at least one child has been killed by chelation treatments[iii]. The complete absence of data in the scientific literature is telling.

There has been no offered explanation as to how a toxin (mercury), could cause the most characteristic symptom of ASDs, larger brains, with a larger number of neurons. There is little similarity between symptoms of mercury toxicity diagnosed by measurement of elevated mercury levels and the symptoms of ASDs.[iv]

The paper that completely disproves the “mercury causes autism” idea is actually one by proponents of the idea[v]. This paper shows a very strong inverse association of mercury with ASD symptomology. That is, people with the most severe ASD symptoms had significantly (p < .0000004) lower mercury levels in their first haircut baby hair. The highest Hg level measured was in one of the normal controls, which was ~100 times higher than the average of the most severely affected individuals (19 ppm (n=1) vs. 0.21 ppm (n=24)). There were >10 most severely affected individuals with Hg levels less than 1/100th that of the highest control (by eye from their fig. 2.). The ad hoc “explanation” that is given, that people with ASDs are “non-excreters” of mercury is completely non-physiologic, and is unsupported by data in that paper, or published before or since. If true, it would contradict much that is well known about mercury physiology.

In the body, Hg and CH3Hg are mostly transported as conjugated thiol and disulfide complexes. That includes complexes with cysteine, homocysteine and glutathione[vi]. These Hg complexes are then ported into cells either with the large neutral amino acid transporter (LAT), or the organic anion transporter (OAT)[vii]. One reason the kidney is a target organ is that much of homocysteine metabolism occurs there and Hg is transported as a homocysteine disulfide.[viii],[ix]

Once the Hg is in the kidney, it causes the expression of metallothionein which sequesters Hg. Typically, Hg is taken up rapidly by the liver, but over the course of several days, after the kidney expresses metallothionein, the liver loses Hg and the kidney takes it up.[x] , [xi] CH3Hg in vivo is slowly oxidized to Hg(0).[xii] The major excretion pathway is via the stool. Bile salt anions and thiols (glutathione GSH) are co-excreted in the bile.[xiii] I suspect that thiol mercury complexes are excreted in the bile via the OAT1, which excretes bile anions and also mercapturic acids[xiv], and then enteric bacteria break the complex, then demethylate it with the Hg(0) being bound to something (porphyrin?, sulfides?) or not reabsorbed. In any case, bile excretion is coupled to GSH excretion. Conjugation of GSH to xenobiotic (and normal metabolites) is an important pathway for detoxification and excretion. If this pathway were disrupted by several orders of magnitude (necessary for the “non-excreter” idea), no doubt there would be effects (jaundice, fat malabsorption, liver failure, gall stones?).

In utero (guinea pig), Hg is primarily located in the fetal liver bound to metallothionein. This Hg then redistributes in neonates.[xv] Metallothionein is normally used to store Zn, with Zn being the most abundant transition metal in proteins (~300 Zn containing proteins plus 900 Zn finger transcription factors) after Fe. Interestingly, NO is involved in the mobilization of Zn from metallothionein and its association with the right protein.[xvi] Selenium is important in coupling the redox states of metallothionein and GSH with Zn release and thiol oxidation state[xvii].[xviii] Hg level of hair grown in utero may have a different relationship of Hg(hair) to Hg(total) to Hg(serum) than in the neonate or adult due to the different levels of metallothionein in the fetal liver and different fetal redox state. Metallothionein holds Hg much more strongly than it does anything else. It is likely that metallothionein is the most stable Hg species in the body. Metal ions in MT are coordinated to multiple cysteine residues (4 for Hg, Cd, Zn and 3 for Cu). The stability of metallothionein metal complexes is Hg> Cu > Cd > Zn > Ni.[xix] The absolute binding constants for these metals are: l019 to 1017 for copper, 1017 to 1015 for cadmium, and 1014 to 1011 for zinc.[xx] The binding constant for mercury is likely >1020, but how much greater is uncertain.

Metallothionein expression is increased under conditions of low folate,[xxi] under conditions of oxidative stress,[xxii] and under conditions of heavy metal exposure.[xxiii] Different organs require different levels of different metals to induce metallothionein. Basal expression of MT RNA is higher in the brain, but in the brain isn’t much changed by exposure to metal (Cd). In contrast that in the kidney is somewhat lower initially, but can be 10x higher than brain post Cd exposure.

Hair is primarily crystalline keratin fibers dispersed in an amorphous matrix.[xxiv] The keratin is cross linked by ionic and disulfide bonds. Keratin has a high cysteine content. Hg is trapped both as CH3Hg and as Hg2+. There has been significant work on the speciation of Hg in hair that has reliable demonstrated this. Interestingly, there is a significant correlation with hair color, with dark hair tending to have a significantly higher Hg content than light hair. I haven’t seen a good explanation for this, but there is also a tendency for hair Hg levels to be higher at higher latitudes. This may be due to photochemical reduction of Hg in the hair, followed by its release after deposition.

Hair can pick up Hg from the environment, but once there it is likely pretty stable, so long as the hair is not treated too “harshly”. Treating hair with commercial artificial waving agent (probably derivatives of thioglycolic or thiolactic acid) removed significant (~30%) Hg with each treatment (total of 3 treatments tested).[xxv] Artificial waving agents break the RS-SR double bonds, either via raising the pH, or by using thioacids. A high pH shampoo with EDTA will likely extract some Hg from hair once the R-S-Hg-S-R bonds have been broken. It is doubtful that much Hg would be extracted from crystalline keratin, but it might be from the amorphous matrix. The two different pools likely have the same Hg content when formed (derived from the R-S-Hg-S-R ported in via amino acid transporters as substrate to make keratin. The stability constant for Hg-EDTA (3x1021) is greater than for Hg-cysteine (2.5x1014). It is unlikely that first haircut baby hair was subjected to thiol waving agents or harsh shampoos with high EDTA, so low Hg content reflects low blood Hg while the hair grew. There have been many studies showing moderate to high levels of Hg in hair, 5, 10, 20 PPM among people who eat lots of fish. There is excellent correlation between Hg levels in cerebellum vs. hair (R=0.853) and cerebellum vs. liver (R=0.873), n=39.[xxvi]

Much of the “evidence” supporting a “mercury causes autism” idea is the anecdotal observation by some parents, that their child’s autism was promptly “triggered” by a vaccination, and presumably the vaccine contained thimerosal. However mercury poisoning is almost never acute, unless there is a very large dose (the dose of thimerosal from a single vaccination is not large, and is expected to have essentially non measurable physiological effects). In most of the industrial mercury poisoning episodes, there is significant exposure to mercury (as measured retrospectively in hair) long before there are any symptoms.[xxvii] In the exposure of Iraqis to CH3Hg in grain made into bread, latencies of weeks to months were observed. In the case of a heavy metal researcher exposed to a lethal dose of dimethyl mercury, she exhibited no symptoms for 5 months, however hair growing during those 5 months had extreme levels of mercury, at times exceeding 1000 ppm[xxviii] (over 5,000 times greater than what was observed in the “most affected” ASD group in the first hair cut baby hair study), yet she exhibited no symptoms. It was estimated that she was exposed to about 1344 mg Hg as 0.44 mL dimethyl mercury. The quantity of Hg in vaccines due to thimerosal is variable, about 12.5 to 25 μg per dose. The woman who experienced no symptoms for 5 months was exposed to about 100,000 times more. The promptness of any purported symptoms is strong evidence that those symptoms are not due to mercury. A more plausible “cause”, is immune system activation, the eliciting of which is the reason that vaccines are administered, and which is very prompt. Immune system stimulation alone can have prompt fatal consequences through anaphylaxis, which was an early complication of vaccine usage before modern Good Manufacturing Practices (GMP) and antibacterial preservatives such as thimerosal.[xxix]

I suspect that the lower Hg levels in baby hair of those with most severe ASD symptoms reflect greater fetal and neonatal liver metallothionein due to increased oxidative stress in utero. This might even be reflected in a reduced cord blood Hg concentration. In sheep, in utero, the Hg concentration of liver and kidney due to elemental exposure (amalgam) is ~10x that of brain.[xxx] It looks like a “steady state” Hg uptake was reached. I consider it very unlikely that there is some unknown protein in the brain that is sequestering Hg better than metallothionein (no such protein has ever been shown to exist), and that the sequestering of Hg adversely affects the function of that hypothetical protein such that it causes autism. Metallothionein binds Hg via coordination to 4 sulfur atoms. What type of coordination would a hypothetical protein require to achieve greater stability? Mercury doesn’t usually coordinate to more than 4 atoms. A protein where mercury coordinates to selenium? To multiple selenium atoms? Proteins are only synthesized under the direction of RNA. Presumably children inherit DNA to make such a protein from their parents. If there was maternal expression of such a protein, the Hg would never make it to the placenta. Presumably such a hypothetical protein couldn’t be on the Y chromosome because females get ASDs. If such a hypothetical protein were important in ASDs, it would have been identified in the many genetic studies that have been done.

For ASD individuals to be “non-excreters” of mercury there would need to be a disruption in cysteine and GSH metabolism sufficient to allow 3 or 4 orders of magnitude difference in the blood/brain/hair partitioning for ASD individuals vs. non-ASD individuals. No individual with a correlation several orders of magnitude necessary for the “non-excreter” idea has ever been reported in humans, or animals. Such an extreme disruption in essential amino acid physiology with no other apparent effects seems extremely unlikely. Amino acid transport is extremely well conserved. There are multiple and redundant pathways for amino acids to be transported in and out of cells. Hair is a minor Hg excretion pathway. If hair cells lacked amino acid transporters for cysteine, hair could not grow. If the ASD individual lacked such transporters, such a defect would either be fatal in utero, or would preclude the placenta (which is 100% fetal derived), from absorbing Hg from the mother.

In any case, if mercury were confined to the brain such that it is non-measurable in other tissue compartments such as blood, it could not cause ASD symptoms in other organs.

There is a significant correlation in the baby hair study between maternal anti-D prophylaxis and autism. Anti-D prophylaxis is used to reduce hemolysis of fetal blood cells due to rho D antibodies which pass the placenta. Despite use of anti-D prophylaxis, there is still increased hemolysis as indicated by hyperbilirubinemia[xxxi]. Hemolysis causes release of hemoglobin into the plasma, where it is ~650 times more effective at destroying NO[xxxii] than the equivalent Hb level in RBCs. Hemolysis during the first trimester during neurulation and other early neurodevelopment events could perturb NO physiology without hyperbilirubinemia being present at birth. The correlation may relate to the correlation between Rh incompatibility and oxidative stress (low NO), rather than to the minimal amount of mercury present in the anti-D prophylaxis vaccine.

Jaundice at birth is significantly more common among autistic children than controls, as is RH incompatibility[xxxiii]. I suspect that “subclinical” hyperbilirubinemia may be important too. The relevant development time may be the first trimester. Hemolysis then might reduce NO levels at critical development times, and then be unapparent at term. The low NO causes oxidative stress, which increases expression of metallothionein and greater sequestering of Hg in the liver and kidney. This might be the cause of the decreases in levels of Fe, Zn, and Cu which have been anecdotally reported. However, the level that is important is the level in the target organs, not necessarily the blood. For Fe, blood is a significant “target organ” where hemoglobin contains most of the Fe. But mitochondria also contain lots of Fe. NO has been shown to be important in Fe, Zn and Cu metabolism.[xxxiv],[xxxv] This may be the mechanism for anemia in autism[xxxvi], a consequence of oxidative stress induced metallothionein upregulation. Correcting the low Fe, Cu, and Zn levels therapeutically should be considered carefully. If the underlying oxidative stress is not corrected (by raising the NO level), then increased transition metals may make it worse.

The one study that purports to show elevated mercury levels in ASDIs, actually does not.[xxxvii] This “study”, was a report of children some with ASDs, some without, whose parents presented them to the International Child Development Resource Center for chelation. The “cases” were children diagnosed with ASDs, the “controls” were children with no known ASDs, and no known exposure to mercury who were presented for chelation by their parents (who presumably were willing to pay out of pocket). Testing of blood levels of mercury was not done, the only “testing”, was on urine after the 9th dose of meso-2,3-dimercaptosuccinic acid. The only “data” that is presented, is from a statistical analysis of the results. For the cases (221), the low, mean, high and standard deviation were 0, 4.06, 58.65 and 8.59 μg/g creatinine. The standard deviation is considerably higher than the mean, showing a highly skewed distribution. For the 18 “controls”, the levels were 0, 1.29, 6.2 and 1.54. Still a highly skewed distribution.

I requested the raw mercury levels from the corresponding author so I could do my own statistical analysis and my request was never acknowledged or complied with.

For the mean Hg level to be 4.06 μg/g, and with a high of 58.65 there had to be many (actually most) with levels below 4.06. What change to a body burden does that actually represent? If we assume the average child is 20 kg, normal creatinine excretion is 15 to 25 mg/kg/day.[xxxviii] That would make about 400 mg creatinine/day for a 20 kg child. The two levels of mercury excretion per day are then 1.62 and 0.52 micrograms/day. The difference is 1.1 microgram. Over 3 days, that is 3.3 micrograms. Remember, the average excretion was less than that, and some had no measurable excretion, that is 0.

It is interesting that the mercury excretions discussed in this publication were considered important enough by the authors to publish, yet there was no mention of the resolution or improvement of any ASD symptoms in this, or in any other publication before or since. Demonstrating resolution of ASD symptoms would of course be much more important than demonstration of (a clinically unimportant) increased excretion of a single microgram of mercury, the average amount of mercury in 10 grams of canned light tuna (n=347)[xxxix] (or 1 ounce of tuna over 3 days). Their use of the term “significant”, may accurately denote statistical significance is misleading in that it can easily be misinterpreted to mean clinically significant which it is not.

In short there has been no data in the literature to support the idea that “mercury causes autism”, and abundant data to show it has no merit. It is time the proponents of it stopped subjecting children to chelation, to “remove” mercury which isn’t present, before another child is uselessly killed.

[i] Bernard S, Enayati A, Redwood L, Roger H, Binstock T. Autism: a novel
form of mercury poisoning. Med Hypotheses. 2001;56:462–471.

[ii] Ip P, Wong V, Ho M, Lee J, Wong W. Mercury exposure in children with autistic spectrum disorder: case-control study. J Child Neurol. 2004 Jun;19(6):431-4.

[iii] Sinha Y, Silove N, Williams K. Chelation therapy and autism.
BMJ. 2006 Oct 7;333(7571):756.

[iv] Nelson KB, Bauman ML. Thimerosal and autism? Pediatrics. 2003 Mar;111(3):674-9.

[v] Amy S. Holmes, Mark F. Blaxill, and Boyd E. Haley. Reduced Levels of Mercury in First Baby Haircuts of Autistic Children. International Journal of Toxicology, 22:277–285, 2003.

[vi] Tracey A. SIMMONS-WILLIS, Albert S. KOH, Thomas W. CLARKSON and Nazzareno BALLATORI. Transport of a neurotoxicant by molecular mimicry: the methylmercury–L-cysteine complex is a substrate for human L-type large neutral amino acid transporter (LAT) 1 and LAT2. Biochem. J. (2002) 367, 239-246.

[vii] Rudolfs K. Zalups, Amy G. Aslamkhan, and Sarfaraz Ahmad. Human organic anion transporter 1 mediates cellular uptake of cysteine-S conjugates of inorganic mercury. Kidney International, vol. 66 (2004), pp 251-261.

[viii] Christy C. Bridges and Rudolfs K. Zalups. Homocysteine, System b0,+ and the Renal Epithelial Transport and Toxicity of Inorganic Mercury. (Am J Pathol 2004, 165:1385–1394.

[ix] VERNON T. CANNON, RUDOLFS K. ZALUPS, and DELON W. BARFUSS. Amino Acid Transporters Involved in Luminal Transport of Mercuric Conjugates of Cysteine in Rabbit Proximal Tubule. JPET 298:780–789, 2001.

[x] RUDOLFS K. ZALUPS and JAMES KOROPATNICK. Temporal Changes in Metallothionein Gene Transcription in Rat Kidney and Liver: Relationship to Content of Mercury and Metallothionein Protein. JPET 295:74–82, 2000.

[xi] RUDOLFS K. ZALUPS. Molecular Interactions with Mercury in the Kidney. PHARMACOLOGICAL REVIEWS Vol. 52, No. 1. p.114.

[xii] JOHN C. SMITH AND FRED F. FARRIS. Methyl Mercury Pharmacokinetics in Man: A Reevaluation. TOXICOLOGY AND APPLIED PHARMACOLOGY 137, 245–252 (1996).

[xiii] Bouchard G, Tuchweber B, Yousef IM. Bile salt independent flow during bile salt-induced choleresis and cholestasis in the rat: role of biliary thiol secretion. Liver. 2000 Feb;20(1):27-37.

[xiv] Pombrio JM, Giangreco A, Li L, Wempe MF, Anders MW, Sweet DH, Pritchard JB, Ballatori N. Mercapturic acids (N-acetylcysteine S-conjugates) as endogenous substrates for the renal organic anion transporter-1. Mol Pharmacol. 2001 Nov;60(5):1091-9.

[xv] Minoru Yoshida. Placental to fetal transfer of mercury and fetotoxicity. Tohoku J. Exp. Med., 2002, 196, 79-88.

[xvi] Linda L. Pearce, Karla Wasserloos, Claudette M. St. Croix, Robin Gandley, Edwin S. Levitan and Bruce R. Pitt. Metallothionein, Nitric Oxide and Zinc Homeostasis in Vascular Endothelial Cells. J. Nutr. 130: 1467S—1470S, 2000.

[xvii] Yu Chen and Wolfgang Maret. Catalytic selenols couple the redox cycles of metallothionein and
glutathione. Eur. J. Biochem. 268, 3346±3353 (2001).

[xviii] WOLFGANG MARET AND BERT L. VALLEE. Thiolate ligands in metallothionein confer redox activity on zinc clusters. Proc. Natl. Acad. Sci. USA Vol. 95, pp. 3478–3482, March 1998.

[xix] Kirk B. Nielson and Dennis R. WingeS. Order of Metal Binding in Metallothionein. THE JOURNAL OF BLOLOGICAL CHEMISTRY Vol. 258, No. 21, Issue of November 10, pp. 13063-13069, 1983.

[xx] Dean H. Hamer. METALLOTHIONEIN. Ann, Rev, Biochem. 1986. 55:913-51

[xxi] Wei-Yong Zhu and Peter W. Melera. Metallothionein Is Overexpressed by Hamster Fibroblasts Selected for Growth in 15 pM Folinic Acid and Provides a Growth Advantage in Low Folate1. CANCER RESEARCH 59, 4194–4199, September 1, 1999.

[xxii] Masao Sato and Masuo Kondoh. Recent studies on metallothionein: protection against toxicity of heavy metals and oxygen free radicals. Tohoku J. Exp. Med., 2002, 196, 9-22.

[xxiii] Diane M. Durnam and Richard D. Palmiter. Transcriptional regulation of the mouse metallothionein-I gene by heavy metals. J Bio Chem, 256 (11) 5712-5716, 1981.

[xxiv] LESLIE N. JONES, Hair Structure Anatomy and Comparative Anatomy. Clinics in Dermatology Y 2001;19:95–103.

[xxv] Akira Yasutake, Miyuki Matsumoto, Masako Yamaguchi and Noriyuki Hachiya. Current hair mercury levels in Japanese: Survey in five districts. Tohoku J. Exp. Med., 2003, 199, 161-169.
[xxvi] E. Haca, M. Krzyzanowskib, J. Krechniaka. Total mercury in human renal cortex, liver, cerebellum and hair. The Science of the Total Environment 248 2000 37-43.

[xxvii] Weiss B, Clarkson TW, Simon W. Silent latency periods in methylmercury poisoning and in neurodegenerative disease. Environ Health Perspect. 2002 Oct;110 Suppl 5:851-4. Review.

[xxviii] Nierenberg DW, Nordgren RE, Chang MB, Siegler RW, Blayney MB, Hochberg F, Toribara TY, Cernichiari E, Clarkson T. Delayed cerebellar disease and death after accidental exposure to dimethylmercury. N Engl J Med. 1998 Jun 4;338(23):1672-6.

[xxix] Wilson GS. The Hazards of Immunization. New York, NY: The Athlone Press; 1967:75-84. (cited in Thimerosal in Vaccines,

[xxx] M. J. VIMY, Y. TAKAHASHI, AND F. L. LORSCHEIDER. Maternal-fetal distribution of mercury (203Hg) released from dental amalgam fillings. Am. J. Physiol. 258 (Regulatory Integrative Comp. Physiol. 27): R939-R945,1990.

[xxxi] A Maayan-Metzger, T Schwartz, J Sulkes, P Merlob. Maternal anti-D prophylaxis during pregnancy
does not cause neonatal haemolysis. Arch Dis Child Fetal Neonatal Ed 2001;84:F60–F62.

[xxxii] Xiaoping Liu, Mark J. S. Miller, Mahesh S. Joshi, Halina Sadowska-Krowicka, David A. Clark, and Jack R. Lancaster, Jr. Diffusion-limited Reaction of Free Nitric Oxide with Erythrocytes. THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 273, No. 30, Issue of July 24, pp. 18709–18713, 1998.

[xxxiii] Naya Juul-Dam, Jeanne Townsend and Eric Courchesne. Prenatal, Perinatal, and Neonatal Factors in Autism, Pervasive Developmental Disorder-Not Otherwise Specified, and the General Population. Pediatrics, 2001;107;63.

[xxxiv] Ralph N. Watts and Des R. Richardson. The mechanism of nitrogen monoxide (NO)-mediated iron mobilization from cells. Eur.J.Biochem. 269, 3383–3392 (2002), FEBS 2002.

[xxxv] Masaru Shinyashiki, Kenneth T. Chiang, Christopher H. Switzer, Edith B. Gralla, Joan S. Valentine, Dennis J. Thiele, and Jon M. Fukuto. The interaction of nitric oxide (NO) with the yeast transcription factor Ace1: A model system for NO-protein thiol interactions with implications to metal metabolism. PNAS, March 14, 2000, vol. 97, no. 6, 2491–2496.

[xxxvi] Latif, A. Heinz, P. Cook, R. Iron deficiency in autism and Asperger syndrome. Autism 2002, Mar, 6(1):103-14. (abstract)

[xxxvii] Jeff Bradstreet, David A. Geier, Jerold J. Kartzinel, James B. Adams, Mark R. Geier. A Case-Control Study of Mercury Burden in Children with Autistic Spectrum Disorders. Journal of American Physicians and Surgeons Volume 8 Number 3 Summer 2003 76-79.

[xxxviii] Normal Reference Laboratory Values. N Engl J Med, 1998; 339: 1063-1072.

[xxxix] US FDA Mercury Levels in Commercial Fish and Shellfish ( (accessed 03/26/07)

The focus of my blogs for a while

I will be posting about my low NO hypothesis of ASDs for a while. I am in the process of writing it up, and have been for several years. It is a work in progress, and it is not small. It is the combination of a couple things, a paper discussing the ongoing symtoms associated with ASDs as being caused and exacerbated by low NO which I prepared for Ferid Murad, and a paper discussing how ASDs are an evolved "feature", that invokes the "tool making and using phenotype", the classic nerd with Asperger's (ie me). It is about 32,000 words now, and I estimate it will be over 50k by the time I finish. It is a work in progress because I am still learning and formulating my understanding of this.

Some may consider this to be "woo-like". It is not.

The objective of this paper is not to “prove” that low NO causes autism, but rather to demonstrate that considerable ASD symptomology is consistent with low basal NO, and that there are plausible mechanisms by which low NO could lead to the neuroanatomy and other symptoms characteristic of ASDs. My purpose is to stimulate the interest of ASD adults, and people working with ASD children and adults sufficiently to consider first steps leading to a clinical trial (where the only real proof lies) aimed at increasing that basal NO level. While all treatments have “side effects” which must be matched against any therapeutic effect, I consider the reestablishment of a normal commensal relationship with these bacteria akin to other aspects of “normal” health, a nutritious diet, moderate exercise, sufficient sleep. While there are “side effects” associated with a nutritious diet, moderate exercise and sufficient sleep, there are no known adverse side effects, and it is likely that there are none. Even a modest improvement or amelioration of even a few symptoms associated with ASDs would be sufficient to balance a treatment of negligible risk. I think the improvement may be considerably greater than that.

This document represents several years of work, as I have added to my knowledge of NO physiology and of ASDs. I apologize for it being so long and cumbersome, but NO physiology is not simple, neither are the developmental effects that lead to ASDIs.

Personal note: I have found that increasing my own basal NO levels through the application of AAOB to my skin, has lessened the severity of some symptoms associated with ASDs. My Asperger’s has gotten “better”. In particular my ability to “multi-task” and to work in the presence of distracting stimuli has diminished. I used to be able to read, think, and work productively at high levels while listening to National Public Radio. I was surprised when it became too distracting. However, there has been a marked increase in my ability to absorb, process, integrate and use information and I can now think “bigger” and more complex thoughts and understand things in ways I did not before. My reading speed has increased remarkably (it was never slow). I understand a great deal more of the body language of individuals around me (but as yet find my own body language to be unsatisfactory). I think I am more creative than I was before. This is an unsatisfactory way to describe the changes that I have observed in my thinking process. There are unquestionably marked improvements, although they are subjective. However acquaintances have noticed and commented upon positive changes in my affect and ability to interact socially and my psychiatrist of ~15 years has noted “decreased anxiety, depression and increased emotional connectedness”[i]. In retrospect Asperger’s explains a great deal of my life history, extreme proficiency at science, honors graduate from MIT, coupled with considerable anxiety, social awkwardness, and vulnerability to being picked on. I consider a “placebo” effect unlikely because I was unaware that I had Asperger’s when I started my NO experiment and I knew very little about autism until after I had noticed changes (the reduced ability to multi-task) and started reading about ASDs. As someone with Asperger’s I don’t have the motivation or the ability to be duplicitous. While I do want to be “right”, it is not to satisfy my ego, it is simply so that my conceptualization of reality corresponds with the reality that actually exists. I am perfectly able to accept that I may be wrong. I don’t think I am. As far as I have been able to tell, everything here is completely consistent with everything that is in the literature (once it is filtered for content that is wrong or mistaken). I do not want notoriety; I have agoraphobia, and don’t like crowds. Being the center of attention is extremely stressful. While this discovery will likely bring me fabulous wealth, making the world a better place for my children (and all children) is more important to me, my “legacy”.

I think the changes have to do with percolation. The brain is a neural network in the near percolation threshold. The sensitivity of the network to change is exponentially realated to proximity to the percolation threshold. The percolation threshold is a true critical point, changes are truly exponential, extremely non-linear.

More important than preventing or treating ASDs, is preventing the low NO and low ATP psychosis that accompanies severe stress, which I think induces the state called going “berserk”. I fear world leaders with control of sufficient nuclear weapons to extinguish the human race made delusional and psychotic by low ATP and “stress” more than anything else. I am 52 years old. If I can experience dramatic changes (though they did take several years, and are still ongoing), I anticipate that children on the autistic spectrum will experience even greater changes, perhaps sufficient normalization for some to constitute what some might call a “cure”.

The notion of a “cure” in ASDIs is problematic because of the fear that people who are “normal” would impose their standards of “normality” on ASDIs, and erase characteristics which those ASDIs feel are important aspects of their personalities. While I feel that fear is well grounded (in that erasing abnormal traits (and individuals) has been a very common aspect of human history), raising a person’s NO level will not have that effect. If the “disease” state is an arbitrarily defined one (as I feel ASDs are), crossing the arbitrary threshold constitutes a “cure”. Just as if 5’3” was defined as “short”, becoming 5’4” would make one “tall”. The changes have not made me “normal”, but they have greatly improved my functionality. While an n of 1 does not constitute “proof” it is “data”, (albeit of low statistical power). While my functionality as I perceive it has increased greatly, I suspect that NTIs would still consider me abnormal and in need of “something”. The inability of some NTIs to accept ASDIs is about NTIs, not about ASDIs.

As someone with Asperger’s, the degree of difficulty in getting my ideas even considered by others in the scientific community has been astounding to me. It is astounding because the difficulty has had nothing to do with facts or logic, but rather by the inability of NTIs to understand my ideas. That difficulty has been important in formulating my understanding of the importance of social isolation in the ability to recognize which scientific and cultural paradigms are “wrong” and so be able to go beyond them. Everyone who has rejected my low NO hypothesis has done so without understanding it, no one who has rejected it has (so far) been able to (or willing to) point out any flaws, or inconsistencies, or where it is incompatible with data in the literature. There have been some senior scientists who have accepted my ideas because they have taken the time to understand them (at least in part). When I have (rarely) submitted manuscripts they have been rejected as “uninteresting”.

I suspect that the difficulty NTIs (those without a significant level of ASDs), have in working and acting outside existing cultural and scientific paradigms relates to mirror neurons and the compulsion that NTIs have to be an accepted and acceptable member of society. I suspect that a large part of the antipathy and malevolence that many NTIs feel toward ASDIs (though they may deny such feelings), relates to the absence of a compatible mirror neuron mediated cultural/communication paradigm in the ASDI. I suspect that lack of compatibility leads to an invocation of the “xenophobia subroutine” directed toward the culturally incompatible individual. When a parent feels xenophobia toward their own child, the result is dissonance which at times is resolved by displacing the malevolent feelings onto any perceived “cause”, the evil pharmaceutical industry, anti-chelation skeptics, or in times past witches. Because ASDs don’t invoke mirror neuron mediated recognition, and they don’t pass the subconscious “Turing test”, they don’t appear as “fully human”, and so extreme measures are acceptable to “correct” this (and so make them "human"). These “treatments include chelation, Applied Behavioral Analysis (ABA), and providing negative “consequents” (inflicting pain via electric shocks, which in any other context would be called torture). Alan Turing was very likely on the ASD spectrum, and his brilliance in mathematics was essential to the Allies winning WWII. I find it ironic (and very sad), that Turing himself in effect “failed” the “Turing test” with respect to the British authorities, who then subjected him to hormone injections to “cure” his sexual orientation. It didn’t “work”, and he killed himself.

For every group that has been discriminated against, ethnic groups, women, gays, Jews, cultists, atheists, and now ASDIs, that discrimination has been “justified” by the feelings of the perpetrators that such bad treatment was appropriate because individuals in the group were “different”, and so not fully “human”, and worthy of the natural respect that “humans” are naturally endowed with. In the US Declaration of Independence, the writers state “We hold these truths to be self-evident, that all men are created equal, that they are endowed by their Creator with certain unalienable Rights, that among these are Life, Liberty and the pursuit of Happiness”. What they actually “meant” was that all white land-owners were created equal. The decision to exclude women, blacks, native Americans, and others doesn’t appear to have been conscious, otherwise they would have explicitly done so in their language. The writers of the US Declaration of Independence were not stupid, or ignorant. They were educated and literate. Presumably it was something they simply didn’t have the capacity to understand, much like Louis Armstrong’s statement about Jazz, “Man, if you have to ask what it is, you'll never know.” How many of our own “self-evident” truths are only “self-evident” because of our blindness that there are alternatives?

[i] #### #######, MD., PhD. Director Psychotherapy training Boston Psychoanalytic Society and Institute (personal communication).

Saturday, March 24, 2007

Al Gore and the fever analogy

A number of bloggers have called Al Gore to task for using the metaphor of a baby having a fever to suggest appropriate action to global warming.

I admit that “fever” is a poor analogy, because far too many fevers are of benign origin. A more appropriate analogy would be hyperpyrexia, or malignant hyperthermia, a condition that is virtually always fatal if left untreated. Like global warming, malignant hyperthermia is not well understood, and can occur suddenly, and the response of those ignorant of it will be to repeat the GWB mantra “no one could have predicted…” (I think malignant hyperthermia relates to insufficient basal NO, and insufficient mitochondria leading to (in the case of acute respiratory distress syndrome (for example), excessive expression of iNOS) inhibition of mitochondria leading to excessive mitochondrial potential and expression of uncoupling protein and collapse of ATP levels with the mitochondrial potential being uselessly dissipated as heat, but that is a story for another blog.)

The “problem”, is that by calling global warming a “fever” (a misdiagnosis by a lay person (Al Gore)), other lay people (non-atmospheric scientists and those who don’t understand global warming) think it is benign, like most "fevers". Al Gore should have likened global warming to malignant hyperthermia. A condition that at one time was nearly always fatal in a hospital setting. Now, with better understanding and sufficiently prompt and vigorous supportive treatments, malignant hyperthermia is only about 5% fatal.

Perhaps if we act vigorously and in the proper way(s), global warming will only have a 5% fatality rate. Unfortunately, if we proceed as if “no one has predicted” any bad consequences, by the time those consequences become obvious, it will be too late.

CO2 in the atmosphere right now, is higher than it has been in the last 10 million years. At which time Greenland had very little ice. Under present conditions, Greenland is unstable and will melt. The only question is when, not if. When Greenland melts, sea level will go up 7 meters.

Current “models” of how fast Greenland is going to melt are woefully inadequate. Sort of like the “models” of hyperthermia 100 years ago. That was before the understanding of how medications lowered fevers:

Before the understanding that sometimes fevers accompanied heart failure:

When ice sheets melt, they do so catastrophically. They don’t just sit there and slowly melt like an ice cube (the only model that is understood, and so is the only model that can be used).

I have been doing a lot of research on global warming and have looked into ice melting dynamics.

The models are extremely conservative. What that means is that they only "model" stuff they understand well. If it isn't "understood", it simply gets left out of the model.

What is left out of the ice melting models are the non-linear things that can greatly accelerate the movement of ice into the ocean. These are simply ignored. Not because they are thought to be unimportant, but because it is not known how to model them.

I will discuss several of them. One of the biggies comes from the fact that water is denser than ice. Ice sheets are only stable because they are below freezing from the top, all the way to the bedrock that they sit on. The center of the Earth is hot, and heat only flows down a temperature gradient. So there is a heat flux at the bottom of the ice, from the bedrock into the ice. That heat can only go somewhere if the somewhere it is going is colder than where it is. If the heat can't go anywhere, it accumulates, and the temperature goes up until the ice melts.

When ice melts, the water it becomes is denser, so it sinks to the bottom, and the ice floats in it. If the water layer at the bottom of the ice gets thick enough, you don't have an ice sheet any more, you have a river covered with ice. The ice may be 3,000 meters thick but if the water layer is "deep enough", the whole shebang will flow to the sea.

What keeps the ice frozen, is the heat flux during the winter when the top is very cold, -40 or so. The heat from the Earth flows down the temperature gradient and is radiated off into space. So at the base of the ice, the temperature is near freezing or below, and it gets progressively colder as you go up, until at the very top there are seasonal fluctuations.

What happens when there is melting water at the top? Small amounts can run off. If there is more, what happens? Because water is denser, the pressure at the bottom of a column of water is higher than ice of the same thickness. The ice deforms, the water flows down. All the way down, until it meets ice that is colder than the freezing point of water, then the water freezes, depositing its heat of melting and raising the temperature of that ice to the freezing point.

But that disrupts the temperature gradient that is transporting heat from the base to the surface. If there is no gradient, geothermal heat can’t go any where. So the temperature of the ice sheet goes up. As ice gets closer to the melting point, its mechanical strength goes down. It is less able to withstand shear and stay as a solid mass.

So what happens is that the ice sheet gets warmer, but doesn’t change much in appearance. Until the warming reaches a critical point where the ice isn’t strong enough to support its own weight. Then it starts to move. When the ice is moved by gravity, its gravitational energy is dissipated as heat at the base where there is relative motion. This is why ice is slippery. Frictional energy dissipated at the surface causes melting and a film of water that lubricates the interface. When the base is at the melting point, any additional energy causes melting, and there is no temperature gradient for the heat to flow away to, for the water to freeze again.

It is likely that the melting of Greenland will occur in catastrophic events. It won’t really “melt”, what will happen is that large masses of ice will flow into the sea. That will raise sea level abruptly. How abruptly? Good question. What is going to slow it down? Once it starts to flow, and has a sufficient film of water to lubricate the flow, not much. We could see large chunks of Greenland slip into the sea, 10’s of percent of the Greenland ice mass. 10% of Greenland would raise global sea level by about 2 feet. If that ice flowed into the Atlantic in a week, the rise in the Atlantic would be more than that, and then it would subside. Would there be big waves? I don’t know, I don’t think so, but a transient of 10 feet wouldn’t surprise me. It might be like a 10 foot storm surge that lasts for a month or two. Flooding everything on the coast of the Atlantic. The ice would likely make the Atlantic impassible until it melted. Then the light fresh melt water could well shut down the ocean circulation.

If CO2 continues to be added to the atmosphere, it could get to levels that haven’t been seen since dinosaurs roamed the Earth. There was no ice at the poles then. When the West Antarctic ice sheet melts, that will also add about 7 meters to sea level. When the East Antarctic ice sheet melts, that will add about 60.

Something like 1/3 of humans live below 100 meters altitude. When sea level goes up, where are they going to move to?

Al Gore is trying to tread a very fine line, portray what might happen in a “sound bite” that is short enough that the media will put it on the news, simple enough that some people will understand it, and yet somehow connects to global warming and the need to do something.

Wednesday, March 21, 2007

Introduction, motivation,

The title comes from a quote by (I think), Arthur C. Clarke, something to the effect that the Universe is not only stranger than you imagine, it is stranger than you can imagine.

I consider that my creativity and my imagination are my strongest features, and as an inventor of Mythic proportion, they are of Mythic proportion too.

As I mentioned, this blog will mostly be about my 2 projects to save the world. I anticipate that most people will have difficulty following what I write about, and will think that I am "mad", or grandiose, or both. I am neither. To paraphrase Orac, a statement of fact is never grandiose.

I have been thinking about what most people think about when they think of a "mad" scientist, such as Dr. Frankenstein, someone who had sufficient scientific mojo to raise inanimate objects from the dead. That is not a trivial undertaking. It is a task of extreme difficulty, even beyond my own capabilities. In that kind of scientific undertaking there is no room for self-delusion, no room for mistakes, no room for any error at all. Dr. Frankenstein's conceptualization of reality must have been absolutely accurate and precise, down to the last detail. The characterization of him as "mad", is the common characterization by those who don't understand, of those who cannot understand, those who will not even try to understand. People who are afraid of, and so demonize everything that is outside of their experience.

I think that the key to this world view involves mirror neurons. If your mirror neurons are too robust, the thoughts you can think are greatly limited by them, and only with great difficulty can you escape their stultifying influence.

I close with a quote.

Our knowledge can only be finite, while our ignorance must necessarily be infinite.
-- Karl Popper