Chronic Fatigue Syndrome May Bea Consequence Of

Chronic Fatigue Syndrome, And A Range of Other Illnesses May Be A Consequence Of Persistent Environmental Pollution By Hexachlorobenzene David Grant B.Sc, M.Sc., Ph.D. Ashbank, New Deer, Turriff, AB53 6UX UK SUMMARY The action of heavy metal and organic persistent environmental pollutants (POPs) which include polybrominated and polychlorinated aromatic substances have been long suspected to have a role in triggering of a range of illnesses in susceptible individuals. An extensive literature survey suggests that the major Persistant Organic Pollutant (POP) which should be of major current concern for potential harm to human and animal health is hexachlorobenzene (HCB), a potential pre-dioxin and intrinsic dioxin-like substance which, with its metabolites, has been reported to induce potential multiple organ intoxication effects, including thyroid dependent immune system dysregulation which may be relevant to the aetiology of Chronic Fatigue Syndrome (CFS), and related illnesses as well as a range of other illnesses including foetal neurological development alteration, childhood obesity, diabetes and breast cancer in women. An increase in poorly understood CFS-like illnesses seemed to have started with the 1934 Los Angeles CFS-like illness epidemic and thereafter the tendency for such illnesses to increase can be suggested to suggested to be due to increased global industrialisation leading to high emission of HCB, characteristically formed in chemical plant operations, especially those of the metallurgical and chor-alkali industries and plants producing chlorinated organic solvents, dielectrics, insecticides etc. and also emitted from waste matter incinerator plants. HCB is both a direct and and indirect by-product of the continued industrial production of chlorinated organic molecules. HCB had previously been thought to have been introduced into the environment principally as a result of the use of this substance as a pesticide. [Some of the trade names employed for HCB were Amaticin, Anticane, Bunt-cure, Granox, Sanocide, and Sniedodox]. This and other chlorinated pesticides (including the now largely withdrawn 1,1/(2,2,2-trichloroetylidene) bis(4-chlorobenzene) [DDT] and γ hexachlorobenzene [Lindane], and other highly chlorinated agrochemicals [Aldrin, Dialdrin, Chlordane, Mirex and Heptachlor] were suspected to cause immune system dysfunction and related autoimmune illnesses (including the chronic proinflammatory conditions which include CFS-like illnesses). While almost all of the specific substances which has been identified as Persitant Organic Pollutants (POPs) and the manufacture and use of which have been curbed by international agreement are now decreasing in the air environment, but this decrease seems to be much less for HCB which is present in air and in animal lipids especially in polar environments (cf., e.g., UN Environment Programme Chemicals Antarctic Regional Report December 2002). It has often been suggested that the reason for the continued or increasing presence of HCB in the atmosphere is due to the existence of an as yet discovered reservoir (thought e.g. perhaps to be humic

matter in soil). While this may to some extent be the case it is more likely that the current continued environmental pollution by HCB is more likely to arise from the use of municipal, industrial and medial waste incineration. The use of incinerator plants which produce HCB seems to contravene the conditions of the Stockholm convention of 2004. However the authors of this report did not fully take account of the scientific situation relating to a facile, not-deliberate route of HCB production which differs from other POPs in that it cannot be specifically curbed at the designed manufacturing chemical plant stage. The continued expansion of the use of incineration as a method of disposal of municipal waste inevitably creates HCB as a potential by-product together with chlorinated-dioxins, furans and other dioxin-like molecules in fly ash. It is a basic scientific principle that under incineration conditions self-assembly of HCB will occur via the random sorting of atoms in a related manner to the self-assembly processes which gives rise to chlorinated dioxins and dioxin-like molecules. This is a major part of the mainstream scientifically-based concern for public health which arises from pyrolysis of everyday used organic waste materials. Unlike the other former industrial and agricultural poly-halogenated aromatic POP substances which in most countries are now prohibited from use, and therefore are no longer legally manufactured, HCB continues to be produced in large amounts de novo during the incineration of common domestic, medical and industrial products. While most of this production can in principle be removed (especially when incineration is conducted in industrial sized units) by electrostatic precipitation and bag filtration, the fact that HCB, dioxin and dioxin like molecules are still inevitably always produced during the incineration of municipal wastes. This phenomenon needs to be addressed since the public health and safety aspect of such plants should be noted to be critically on the need for a continuous an exceptionally high standard of plant maintenance and management during long periods of continuous operation during which the feedstock may change in unpredictable ways. While the greatest amounts of HCB and dioxins undoubtedly arise from back yard fires and barrel incinerations, the large-scale production potential of municipal incinerators should also be considered as potential sources, especially as the fly ashes almost always contain HCB and a wide range of other highly toxic chlorinated organic including chlorinated dioxins, furans and biphenyls. With increasing use of incineration of domestic waste this source could be now a major global source of HCB which undergoes global distribution followed by enrichment up the food chain. HCB accumulates in human and animal adipose tissues HCB, it should be noted, is only slowly eliminated form the animal organisms. A major problem exists regarding the acquisition and dissemenation of research results and related information between the scientific community and regulatory authorities for the proper evaluation and establishing safe operational frameworks for the safeguarding of public health relating to HCB intoxication because the necessary expertise required for this requires cooperation between numerous sub-groups of academic and applied scientists pure chemists, environmental scientists, chemical engineers, biomedical scientists and public health experts as well as legislators at local, national and international level. The necessary framework for this critically requires to combine input from environmental, ecological and atmosphere specialists, bench chemists, theoretical chemists, chemical engineers and others and who have hands-on experience of the

how the environment can be affected by various classes of chemical substances and knowledge of the mathematical models describing pyrolysis behaviour during (including pre and after burn) temperature conditions present during the incineration of municipal wastes. It would require national environmental agencies to have hands on knowledge of such matters and optimally to operate experimental incineration plants. This is in practice difficult to achieve. The suggested unique environmental hazard posed by HCB is suggested to be reminiscent of the global intoxication of the biosphere by lead from gasoline. HCB will be difficult to remove from the environment and eradicate from the human body to which ends further research is required, perhaps this should include investigation of the ability of anionic polysaccharides especially glycosaminoglycans, to assist with the elimination of HCB stores. INTRODUCTION HCB has unique physical and chemical properties including high hydrophobicity leading to low water but high fatty tissue solubility; the unique thermodynamic and kinetic properties of HCB require that this substance is always formed in large amounts during the pyrolysis of chlorocarbons (1a) chlorinated organic molecules (including polyvinylchloride) as well as non-chlorinated organic wastes e.g. hydrocarbons pyrolysed in the presence of any chlorine atom source (including inorganic materials). It is formed when domestic and industrial waste materials (1b) are burned under common conditions where chlorine can be generated in situ by the Deacon process (1c). Although some well-designed incinerators are believed to be capable of almost total elimination of HCB and other chlorinated and as well as analogous brominated aromatic substances, abundant HCB formation still occurs during waste incineration in such plant some of which is likely to escape (e.g. being adsorbed on the surface of very small soot (carbon) or mineral particles which can escape post-combustion clean-up trains) moss especially when combustion is conducted under suboptimal incineration conditions. HCB can be transported intercontinentally in the troposphere and become enriched in the animal food chain. Although HCB is sparingly soluble in water, it is highly lipid soluble, allowing it to become accumulated in plasma membranes (accumulating in human adipose tissue over many years) and to occur in the fat content of meat and milk. HCB is now a ubiquitous intoxicant in humans (2a,b). HCB apparently promotes a general chronic state of oxidative stress in animals and humans (3). This is thought to lead, in addition to the promotion of CFS and autistic-spectrum disorders, to a range of other illnesses have also been putatively linked to HCB intoxication, viz., alteration in foetal development, diabetes, asthma, osteoporosis, some forms of cancer and multiple sclerosis and has been proposed to be a major causative factor in the current global epidemic of obesity, a key factor in the aetiology of diabetes as well as in thyroid atrophy, liver disease and prostate cancer. HCB seems especially to negatively affect the central and peripheral nervous systems, parathyroid and kidney function and to perturb the general metabolic processes.

Many diseases which are promoted by HCB intoxication seem to arise as a consequence of immune system dysfunctions especially a disruption of the endocrine system and the induction of autoimmune reactions via the effects which HCB exerts on thryroid factors, cytokine and chemokine activities. These interactions, however, seem likely to demonstrate a wide variation in species and individual susceptibility. The effect of HCB on liver intoxication includes the induction of damaging Cytochrome-450 isoenzymes and the disruption of heme biosynthesis which is associated with the promotion of porphyria which can further promote Fenton reaction free-radical promotion and the associated lipid peroxidation damage to tisues. A known adrenal intoxication effect of chlorinated aromatics (3c) might include similar effects of HCB metabolites which could affect the formation of C-11 –OH formation in cortisol which has been associated with osteoporosis. HCB intoxication has also been indicated to negatively affect animal reproductive organs and damage foetal development (which leads to a increased prenatal and postnatal mortality). Although some animal studies and occupational intoxication studies have suggested highly variable rates of risk, many of which would scarcely support the notion of a serious health risk arising from the environmental contamination by HCB, nevertheless the present wide-ranging assessment of the large amount of available data in the scientific literature tends strongly to indicates that the contrary view is more likely to be correct, namely that the a prolonged modest intoxication by HCB could provoke a major systemic, if often subtle intoxication effect which is inter alia responsible for common CFS-like illnesses. Although most animal studies employed ingestion of much greater amounts of HCB than seems appropriate for modelling the general human environmental intoxication situation, damage to primate ovaries has been reported to occur with a dietary intake of 0.1µ g/kg/day which is believed to be the approximate average human ingestion value. HCB seems to be more closely associated than are other potential environmental pollutants with both foetal in infant neurological development. This is thought to be of especial relevant to the aetiology of autism, asthma, chemical hypersensitity, chronic bacterial and viral infections as well as CFS and related diseases. CONCLUSIONS While currently available evidence form human subject is relatively sparse for HCB being the primary causative agent of ME/CFS, it is also evident that there is growing evidence that HCB intoxication could be directly linked as the primarily causative agent to a number of health problems which include the global epidemic of obesity, diabetic disorders, cancers, infant mortality, fertility and neurological damage. Further work is of course needed to confirm the available evidence that HCB is responsible for a range of illnesses the aetiologies of which have previously been uncertain. The current information suggests that this possibility should be addressed specifically. The previous studies of HCB intoxication seem to have been concerned principally with accidental or industrial exposure and the accidental global contamination scenario has been discovered more or less by accident.

Further studies of the role of HCB formation from the incineration of domestic and industrial waste material urgently warrants further study.

SELECTED REFERENCESPHYSIOLOGICAL STUDIES OF HCB INTOXICATION Genetic Dependence of HCB Toxicity Studies of HCB intoxication in the rat support the notion that the observed effects in man arise from the induction by HCB of a chronic pro-inflammatory condition. It was found that rats differ markedly in their susceptibility to HCB intoxication and a situation may also applies to human intoxication by HCB and could suggest a high individual susceptibility variation in human genetic predisposition to ME/CFS arising from HCB intoxication. HCB-like intoxication symptoms can apparently in some circumstances can also be produced by pentachlorobenzene (PCB) which has been widely used as a wood preservative. It seems that pentachlorbenzene is also formed in vivo from HCB. Cf. Van Ommen B Van Bladeren PJ Temmink JHM Muller F (1985) Formation of pentachlorophenol as the major product of microsomal oxidation of hexachlorobenzene Biochem Biophys Res Commun. 126 (1): 25-32 De Meirlier (2000) suggested that CFS arose 10-20 years following exposure to PCB which seemed likely in vivo to slowly create a range of the more toxic dioxin-like metabolites. Carcinogenic Properties of HCB An effect of HCB intoxication on gap junction has been reported. This may be relevant to the gender-specific promotion of hepatic tumours by HCB (Plante et al., 2002). The association of HCB intoxication with human cander includes the study (Charleir et al. 2003) which seems to link blood serum organochlorine residues with breast cancer in women (where the amounts of HCB present in fatty tissues of the breast were correlated with the severity of this cancer). Recent research (Arrebola et al. 2009) also suggests that women are more susceptible to the cancer promoting effects of HCB than are men. A large body of toxicological data for HCB is available from animal experimentation Including a recent study of mild HCB intoxication in the rat (4) (4) Ezendam et al 2004 Numerous studies of HCB intoxication in laboratory animals originating from interest in a human populations which had been exposed to HCB. This began with the large human poisoning incident from 1955-1959 in Turkey which was caused by an

accidental contamination of human food by HCB (which was at that time commonly used as an antifungal agent in agricultural seed). A pertinent finding from these studies was that there seemed to be some additional source of HCB (e.g. from waste incineration?) which seemed also to affect the control subjects. The blood contents of HCB in the Turkish studies seemed accurately to predict the incidence of spontaneous abortion in women of childbearing age. This included both the fungicide-HCB intoxicated group as well as the general population HCBintoxicated group. The very high level of infant mortality in Turkey at the time of the 1955-1959 intoxication seemed to be associated with the high level of HCB intoxication in that country at that time. There are also hints that intoxication by HCB and other chlorinated organic molecules is related to demand for in vitro fertilization (as suggested by a study of Canadian women (2a)). A sub-set of CFS patients (with defined Hughes syndrome) was also associated with a high spontaneous abortion rate but this was apparently alleviated by heparin therapy. It is conceivable, from the known ability of such heparin to upregulate heparan sulphate biosynthesis, that such heparin administration could counter the negative effects of HCB and other dioxin and dioxin-like molecule intoxication on heparan sulphate biosynthesis. An alleviation of the symptoms of CFS by heparin therapy seems to greatly benefit a sub-set of patients (this applies, e.g. to ca. 25% of such patients cf. Andrews 2003 [personal communication]; it should be noted the possibility that HCB intoxication was involved does not seem to have been considered in these cases, however). The damaging effect of HCB intoxication on early embryo development was confirmed by the work of Sharpe & Irvine et al., 2004 (3); cf. also Duanmer et al. 2001). Dioxin-like Activity of HCB in Human Milk While being much less active for binding to the aromatic receptor (AhR) than true dioxins, the much higher amounts of HCB apparently present in the environment makes HCB a likely major contribution to the dioxin-like activity of human milk in agreement with assay results (e.g., Van Bergelen et al. 1998 (4)). It is thought that foetuses and infants may be more at risk of adverse effects of HCB intoxication than are adults. (4) Van Birgilen AP (1998) Hexachlorobenzene is a possible major contributor to the dioxin activity of human milk Environ Health Perspect 106:683-688 Synergistic Interactions Between HCB and Dioxin Toxicity An important synergistic interaction between HCB and dioxin intoxication mechanisms has been indicated, whereby HCB can

dramatically increase dioxin toxicity effects (especially those which lead to thyroid atrophy). The mechanism of this effect is unknown. There are major environmental implications of these researches which, however, seem not to have been continued with. Li SMA Denomme MA Leece B Towner R Safe S (1989)

Hexachlorobenzene: biochemical effects and synergistic toxic interactions with 2,3,7,8-tetrachlorodibenzo-p-dioxin Toxicol Environ Chem. 22: 215-227 Further work is clearly warranted to determine the detailed reaction mechanisms involved which seem not to be known. [Although written some time ago, the introduction to this paper noted that in 1989 it had already been established that HCB had been detected as “a by-product in the production of numerous organochlorine industrial compounds and formed during the combustion of organic waste containing a source of chlorine. HCB, in common with other halogenated aryl hydrocarbons, is highly stable and resistant to chemical, biological and physical degradation. HCB is also highly lipophilic and under conditions of adequate ventilation, HCB will also slowly volatilize. Not surprisingly, HCB has been identified as a contaminant in almost every component of the global ecosystem including fish, wildlife, human adipose tissue, blood and milk. HCB elicits a broad spectrum of species-dependent toxic and biologic responses with include neurotoxicity, body weight loss, porphyria, cutaneous lesion, hepatomegaly and liver damage, nephrotoxicity, splenomegaly, reproductive toxicity, carcinogenesis and immuno-toxicity. HCB also induces several drug metabolizing enzymes including the cytochrome P-450-dependent monooxygenases.” Since 1989 the above details relating to HCB have been further confirmed; in addition HCB toxicity is now thought to promote an even greater range of illnesses than was suspected in 1989, e.g., HCB seems to be a major factor in the induction of breast cancer in women and in the global epidemic of overweight children. -----------------------------------------------------------------------------------------------HUMAN CANCER AND HCB INTOXICATION Higher Risk of HCB Intoxication of Women than Men Arreobola JP et al., (2009) 35 (1): 27-32 Prediction of concentrations of HCB in human adipose tissue : a multivariate analysis by gender in Southern Spain [HCB was measured and quantified in 90.7% in adipose tissues of 387 subjects. It was found that the women in this survey had three fold more HCB than the men. Further work is required to establish the reason for this.] HCB and Breast Cancer in Women Charlier C et al., Occup Environ Med. 2003 60: 348-351 Breast cancer and serum organochlorine residues [HCB appears to occur at higher concentrations in breast adipose tissue of women suffering from breast cancer than control tissues. (There was no correlation between the HCB concentration and oestrogen receptor status). Although these data do not prove that HCB causes breast cancer the authors had noted that previous work had shown that HCB can demonstrate a tumour promoting activity in human breast epithelial cells.] Gender-Specific HCB Induction of Rat Hepatocarcinoma Generation Plante I et al., Toxicol Sci 2005 88 (2): 346-357 Involvement of integrin-linked kinase pathway in HCB-induced gender-specific rat hepatocarcinogenesis [Rats exposed to HCB showed a decreased liver gap junction intercellular communication; this may be part of the reason why HCB is an epigenetic carcinogen]

Intoxication of male industrial workers in Flix, Spain seemed, indeed, to suggest a lesser impact of HCB intoxication on human health than had been suggested by the Turkish studies. (Herreroied et al. 1999 (3e) however later demonstrated in a similar population group, an effect on steroid hormone regulation which might be related to the induction of prostate cancer (Ralph et al 2003 (3c) who also reported that low HCB intoxication levels were associated with an augmented prostate activity while very high HCB intoxication levels caused overt prostate damage. There may have be unknown environmental intoxication differences between individual human populations of other toxic cofactors which could conceivably act to augment HCB toxicity (e.g. less medical intervention or agricultural application introduced mercury intoxication in Spain than in Turkey?). [Cf. also the literature reviews conducted by Jarrell & Gocmen , 2000 (2a) and the US Department of Health and Human Services Agency (2b)]. Promotion by HCB of Fenton Reaction Induced Free Radical Tissue Destruction HCB has specific non-dioxin toxic effects which include the dysregulation of heme biochemistry and release of Fenton Reaction inducing iron ions which can stimulate destructive fee-radical generation as well as protein nitration. These processes can, at least to some extent, be inhibited by certain antioxidant molecules and also, it is believed by a putative additional system of anti-nitrant protection which combats dysregulation of nitric oxide biochemistry which includes actions of glycosaminoglycans. A mechanism by which HCB intoxication induces neurological damage in man and animals seems likely to be via the effects of tyrosine nitration. An example, selected almost at random from scenarios of potential relevance to the aetiology of CFS indicates that an induced adaptive resistance (IAR) to nitric oxide in the central nervous system is related to the heat shock protein HO-1 inhibition of protein nitration; motor neurons from HO-1 null mice showed elevated levels nitrated proteins (Amey Bishop Research group site (5a)). (5a) http://www/uah/colleges/science/biology.amy.amy.htm Tyrosine nitration is also believed to negatively impact on the immune system Iron dyshomeostasis resulting from porphyria is a possible promoter of systemic reactive nitrogen reactive species generation (e.g. through promotion of Fenton-type free radical formation chemistry. Vesely et al (1998) Free hemin: a major source of iron for Fenton reaction induced H-O -1 Am J Cell Physiol. 275: C 1087-C1094 Cf stinulation of liver H-O -1 induced hepatic porphyria Arch Toxicology 1998 72 (6) 355-361 cf. Thomas et al. 2002(6a, cf. 6b). A further nitric oxide imbalance effect will likely be associated with N-sulphated glucosamine-based polysaccharide depolymerisation catalysed

by the activities (e.g. non-enzymic de-N-sulphonation) unlignaded, redox acitve iron ions, the presence of which indicated by blood ferritin levels have bee associated with cardiovascular dysfunction related mortality in Finnish men. Boost of specific anti-nitrants (e.g. by dietary means) might be predicted to show beneficial effects in CFS patients. Chelation therapy for excess redox metals is predicted to diminish the damaging effects of Fenton reaction iron and copper ion augmentation effects. REPORTED DIRECT INVOLVEMENT OF HCB IN THE AETIOLOGY OF Myalgic Encephaolmyelitis/Chronic Fatigue Syndrome (ME/CFS) Environmental pollution by organic pesticides which are known to act inter alia as endocrine disrupting agents in animals and humans of the persistent provides a rational scenario for consideration as a trigger of ME/CFS malaise, since these illnesses appear to include endocrine system dysfunction. The highly persistent chlorinated pesticides include DDT, its metabolite DDE (1,1-dichlor-2,2-bis(4-chlorophenyl)ethane), chlorpyrifos (o,o-diethyl O-3,5,6 trichloro-2-pyridyl phosphorothiolate) or polybrominated flame retardants. However currently more widely distributed and persistent endocrine disrupting environmental pollutant which is now suggested to more likely be the principal inducer of ME/CFS is HCB which seems fairly certain to be currently produced in an uncontrolled manner By the incineration of medial and municipal waste as well as its formation during the manufacture of polyvinyl chloride. HCB, as well as its metabolic precursor Lindane (Gamexene) a benzene hexachloride isomer used in the past (and also continued to be used in some countries) as an insecticide and its metabolite pentachlorophenol have been directly implicated as suggested by several studies, most notably Dunstan et al 1995, 1999, in the aetiology of CFS, Although other toxins and pathological organisms also seem to be contributory factors to these illnesses, currently categorised as Chronic Fatigue Syndromes, the general intoxication of the human population by HCB must, it can be suggested, also contribute to a widespread pro-inflammatory background situation which will inevitably cause, in genetically predisposed populations, a range of illnesses which often might only be slight malaises in many persons but can also progress to major lifestyle diminishing clinically-defined ME/CFS diseases in others. The health effect caused by some non-agricultural source of HCB intoxication in Australia which was indicated by the reports of Dunstan et al. which although on a modest scale, seemed to link CFS directly with HCB intoxication and highlighted the unique role played by HCB in the aetiology of CFS, including the role of the accumulation of HCB with age (which significantly correlated with the blood serum concentrations of HCB in a CFS affected group). [The HCB contamination value of >2ppb was found for 45% in a CFS population group compared with 21% in a non-CFS group. The CFS group overall had a HCB + DDE of 15.9ppb compared with 6.3ppb in the control group. There was a very strong correlated with age and HCB contamination in the CFS group which was not observed in the control groups. This suggests a specific link between HCB intoxication and CFS].

The presence of HCB in the general population who had not been exposed occupationally also could suggest some major overlooked source of HCB. This is likely, as was indicated by the major Turkish intoxication episode (vide supra) to include a major input arising from the facile formation of HCB during burning of common PVC-containing industrial and household waste. Extracellular Matrix & HCB Amongst the tissue protective mechanisms are those provided by the glycosaminoglycans which occur in the extracellular matrices and cell surfaces of adherent cells of all animal species. An important member of the polysaccharide families is the polysaccharide family of heparan sulphates(the heparanome), a major systemic high system manger function provider the established functions of which include tissue protection achieved through the regulation of growth factor, anticoagulant activity, immune responses and antioxidant activities. It is further predicted that relatively poorly understood but probably, heparan sulphate is subject to perturbation by HCB intoxication which includes perturbation by HCB of steroid hormone systems. Since the actions of heparan sulphate biochemistry for regulation of growth factor activities are known to be highly involved in the development processes and the fact that HCB other dioxin-like and dioxin molecules are also known to disrupt the early development of the embryo the mechanism by which HCB impacts on these process is consistent with a perturbation by HCB of heparan sulphate biosynthesis and signalling. The known disruption by HCB of sulphotrasferase activities could suggest that HCB alters the degree of sulphation of heparan sulphate. Heparan sulphate is known to modulate integrin activities which are also known to be involved in gender-specific rat hepatocarcinogensis (cf., Arreboal et al., 2009) Alteration induced in the extracellular matrix component activities as a result of HCB intoxication could be central to the modus operandi of HCB induced carcinogeneisis which could indicate possible therapeutic intervention strategies, e.g., Loose et al. (1981) (cited in 2b) found that HCB, polychlorinated biphenyl and dieldrin significantly diminished fibronectin synthesis. Thyroid dysfunction including that promoted by HCB may be the origin of fibromyalgia (FM) an illness with symptoms which overlap with those of CFS and characterised by the presence of large amounts of the glycosaminoglycan hyaluronan fragment in the blood serum (a phenomenon which has been suggested to be a useful diagnostic test for FM). HCB and dioxins have been found to increase gene transcription of sulphotransferase enzymes which result in diminished steroid hormones (due to an increased rate of their excretion thereby induced). On the other hand the HCB metabolite pentachlorophenol apparently inhibits sulphotranferase activity. C Rimington a key early investigator of HCB toxicity seemed to suggest [in a 1984 letter to the author] by extrapolating from his work on the effect of organochlorines (especially carbon tetrachloride) on liver that glycosaminoglycans might be protect against such intoxication.

Another carbohydrate-based detoxification method applicable to dioxin-like molecules (especially to 2,3,7,8-tetrachlorodibenzo-p-dioxin-induced oxidation stress in mice) seems to be afforded by chitosan (cf. Shon et al. (2002)). Other Detoxification Pathways It is believed that there are two major detoxification pathways of HCB in the liver which are by the mercapturic pathway by glutathione conjugation and urinary production of N-acetyl-S-(pentachlorophenyl)-cysteine and degradation by Cytochrome P450 catalysed oxidative dehalogenation to form pentachlorophenol and 1,4-tetrachlorohydrocquinone (cf. the review by Michielson et al. (1999)) Urinary Metabolites of HCB Include major amounts of pentachlorophenol, 2,4,5 and 2,4,6 trichlorophenol. Cytochrome 450 Specific cytochrome 450 expression is thought to occur via the AhR aromatic receptor with possible systemic side effects, induction of liver microsomal enzymes, altered liver functions (including gluconeogeneis), dysregulation of tryptophan metabolism and induction of (altered Krebs cycle effect) malic acid gene transcription by a thryroid hormone response element mechanism (7). Toxogenomics of HCB in the Rat A large body of toxicological data from HCB is available from animal experimentation studies including a recent toxicogenomics study of mild HCB intoxication in the rate (Ezendam et al. 2004(3a)) which indicates that the systemic effects of mild HCB intoxication are greater than previously thought and highlights the induction of systemic inflammatory responses including gene expression of annexin-1 (an indicator of chronic oxidative stress) which blocks leukocyte migration and induces apoptosis in inflammatory cells, thymus, liver and kidney as well as causes neurological dysregulation and the acetylcholine receptor. A large number of previous studies had shown that HCB is sequestered in membrane lipids and exerts (sometimes subtle but relevant) systemic effects including induction of general oxidative stress as well as specific HCB-induced erythrocyte dysfunction (inhibition of key metabolic enzymes and associated alteration in erythrocyte structural and functional damage). Note a The Van Wazer Structural Reorganization Theory & the Aubrey-Van Wazer Equilibrium A fundamental chemical driving force (hitherto well-known only as an esoteric subject of interest to a small number of pure chemistry researchers) is now suggested to be the scientific reason why an efficient HCB synthesis occurs during common incineration processes when molecules containing carbon and chlorine atoms are incinerated. Where pyrolysis of chlorine free organic matter and also common pure hydrocarbon feedstock can lead to graphite char, a similar pyrolysis of compounds containing C

and Cl leads inevitably to the formation of CCl4, C2Cl6 and C6Cl6 in amounts determined by the ratio Cl/C. Mixed H, Cl and C systems are predicted to loose H/C char (solid separated phase formation) in such a way as to increase the Cl/C ratio in the gas phase and lead to formation of abundant C6Cl6 even when the starting materials consist of mixtures containing lightly chlorinated organic compounds. C6Cl6 is a very stable chlorocarbon due to the stability of the free radial derived from it (intrinsic antioxidant effect) and low water solubility; -(CH2)-n as in high density polyetheylene or in humic polymers is similary highly stable in the environment for the same reasonf for reason. Such compounds demonstrate an inability to form the types of free radicals which engage in rapid oxidative degradation. On the other hand the hypothetical polymer polytetracholorethylene -(CCl2)-n is highly unstable with respect to cyclic rearrangement and dechlorination to form HCB. A further aspect of numerous pyrolysis systems apart form the formation of unusual an unusual major end product in the case of chlorocarbons there frequently occur a background scrambling process to form a wide range of products which for the O/C/H/Cl system includes a wide range of dioxins, furans, chlorinated benzenes, chlorinated biphenyl and numerous other chlorinated aromatic compounds. A wide range of systems of families of chemical compounds were found experimentally to exhibit this kind of thousands of individual molecules. This phenomenon could however be dissected in terms of systematic reversible assemblydisassembly of molecular parts e.g. ortho (isolated) groups, end groups, middle groups, one-way branch groups, two-way branch groups and overall ring/chain system balances. These processes seemed to be more characterised by thermodynamically directly rather than kinetically determined reactions which lead to a Monte Carlo creation of random structure reassemblies. The HCB formation during chorocarbon pyrolysis is a well-defined example of the ‘no mechanism’ (really severe multiple mechanism) chemistry where the amounts of products formed were found to be accurately predicted by the sort of logic mentioned in the previous paragraph for which appropriate software was developed originally found to describe the occurrence of random rearrangements which describe the inorganic chemistry of condensed on phosphorus(V) oxyacid-based oligomers and polymers. The same mathematical model was also found to be applicable to the “pyrolysis” of a range of inorganic polymers including polysulphide, polysulphate and many other types of inorganic and organometallic polymer systems (studied in the laboratories of the Monsanto Company in St Louis) (cf. Van Wazer (1962); Aubrey & Van Wazer, (1962), and numerous studies of Van Wazer with co-workers who included Dungan, Grant, Groeghweghe, Rausch, Mark, Matula and Moedritzer. The chlorocarbon pyrolysis system was further studied and the original Aubrey-Van Wazer process confirmed to be the dominant one by Grant working at I.C.I. Ltd U.K. (Grant (1974) loc. cit. ). More recent research (cf. Taylor & Lenoid, 2001 review) confirmed that the AubreyVan Wazer HCB-process is applicable to a wider range of starting elemental composition mixture than purely carbon chlorine systems and can be applied to the fuller comprehension of how a very large number of individual molecular species (cf. the ca 1000+ range of chlorinated aromatic molecules, including chlorinated dioxins,

furans and biphenyls, which are found to occur in association with fly ash from municipal waste incinerators). The sub-systems which control these events can however be assigned as groups of simpler molecular part exchange equilibria. The Cl/C System Subgroup (A dominant structural reorganization process has been identified as occurring during pyrolysis chlorocarbons in the temperature range including 300-1110oC [the Aubrey-Van Wazer equilibrium] 12 CCl4

+

Aliphatic ‘ortho’ monomer

C6Cl6 [HCB] Middle groups (in predominant most stable ring)

=

9C2Cl6 Aliphatic chains end group

The H/C System Subgroup A structural reorganization occurs similarly to the above aliphatic/aromatic equilibration but this is perturbed by a major dominant phase change process leading from the aromatic ring to char or ‘soot’ formation. The incinerator modelling woks show that HCB is always formed during the pyrolysis of any chlorinated organic material such as polyvinyl chloride, chlorinated humic material or even non-chlorinated organic substances incinerated under conditions where hydrochloric acid can also formed in the presence of transition metal catalysts (especially CuCl2). Although modern incinerators can in principle be designed to avoid major emissions of HCB, HCB will continue to be formed by use of older technology. A problem with the commercial motif for incineration for electricity generation is negated by the high cost of the technology needed to reduce HCB emissions to acceptable levels. This mitigates against using the best available technology. Current pratice seems to ignore routine online analysis for HCB and dioxins instead attempting to conduct incineration ot optimal traditional avoidance of poor combustion indicators (carbon monoxide, carbon particle formation). There have been attempts to use surrogate molecules to justify claims that emissions are safe. The scientific basis of such practices seem to be intrinsically unsound. Although in the use of incineration for the disposal of municipal waste the formation and potential emission into the atmosphere of HCB containing flue gases and deposition of HCB on fly ashes, together with a range of hundreds types of highly toxic dioxins, furans and chlorinated biphenyls and related chlorinated organic molecules is well known to be an intrinsic risk to public health as indicated by the publication of numerous modifications of the design of incinerators in an attempt to limit the release of these substances to the environment. Strategies currently under consideration include e.g. by their recycling to the combustion chamber. Since under slightly suboptimal conditions there is a putatively risk posed to health by emissions of HCB strongly attached (adsorbed) to soot (carbon) nanoparticales (a phenomenon

requiring more research) or deposited on SiO2/Al2O3-rich inorganic particles. For incinerator operation and on-going use of toxic waste dumps to dispose of HCB and dioxin contaminated fly ash also requires further research (e.g. to so stabilise the ash to prevent the leaching from it of dioxins as well as a range of toxic heavy metals that it may be safely used as an additive to Portland Cement or asphalt for road construction) . The emission of unacceptable levels of HCB occur most likely to an unacceptable extent with currently employed older medical municipal or industrial incinerators, back yard and farm barrel incineration, forest fires or as a result of volcanic activity. This could make the eradication of HCB from the environment seems a virtually impossible task. Selected References Jarrell J Gocmen A (2000) A review of human and sub-human primate toxicity of hexachlorobenzene Pure Appl Chem. 72 (6) 1015-1021 Cf. Rimington C et al. (1961) Nervous and biochemical disturbances following hexachlorobenzene intoxication Nature 191: 363-366 (1a) Aubrey NE Van Wazer JR (1964) J Amer Chem Soc. 86: 4380 Cf. Grant D (1974) The pyrolysis of chlorocarbons J Appl Chem Biotechnol. 24: 49-58 [The formation of HCB was confirmed to occur during the pyrolysis of a range of starting material and other intermediate reaction products. The kinetics of the scrambling processes responsible for HCB formation were elucidated. Although the reactions involved in the scrambling processes the outcome of which was determined by the overall Cl/C ratio in the pyrolysis chamber, the reversible processes involved in the Monto Carlo scrambling seemed to occur by different routes in the forward and reverse directions in the empirically derived control equation 12CCl4 + C6Cl6 = 9C2Cl6, hence it seems not to be a true thermodynamic equilibrium but is more accurately described as a pseudo-equilibrium process; the catalytic effects of the attainment of this process of a range of inorganic ions was established]. It is considered that the above references, although they do not deal with incineration of waste as such, deal with the fundamental properties of reorganization of carbon and chlorine and give the greatest insight into the intrinsic problems facing the as yet not successful attempts to safely incinerate industrial and municipal waste.

The reoganization of a wide range of types of chemical substances was studied by the team (at Monsanto St Louis) including organic organometalloid, and organometallic as well as inorganic families of compounds. Later studies (cf. the following references) use different intellectual approaches to rationalize the nature of the reaction processes occurring during incineration. The results presented in these later studies however, support of the Van Wazer stochastic reorganization theory . Taylor PH Lenoid D (2001) Chloroaromatic formation in incinerator processes Sci Total Environ. 260: 1-24 Lenoir D Wehrmeier A Sidhu SS Taylor PH (2001) Formation and inhibition of chloroaromatic micropollutants formed in incineration processes Chemosphere 43: 107-114 Choudhry GG Hutzinger O (1983) Mechanistic Aspects of the Thermal Formation of Halogenated Organic Compounds Including Polychlorinated Dibenzo-p-Dioxins Vol 4 of Current Topics in Environmental and Toxicological Chemistry Gordon and breach Science Publishers [This comprehensive review of the formation of dioxin-like substances (including HCB) begins with the statement “The disposal of industrial and municipal wastes and resides without contaminating the environment is one of the major problems of today”… it goes on to note that “The most promising method for the disposal of this refuse is incineration or burning” and “The gaseous products consist of nitrogen oxides, sulphur oxides, carbon monoxide, carbon dioxide, water, oxygen, ammonia, hydrogen chloride and some hydrocarbons. Fly ash contains 70-95% inorganic matter, While trace amounts of elements such as chromium, nickel, copper, zinc, silver, lead, sulphur, phosphorus and boron have been detected, the predominant elements in fly ahs are silicon, iron, aluminium, calcium , magnesium, potassium and sodium. Polychlroinated dibenzeo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and polychlorinated phenols have beenidentified by Hitziner and his coworkers in the flue gas and fly ash from some municipal incinerators in The Netherlands, Busner et al showed similar identification in samples from municipal incinerators in Switzerland. Recently Karasel and his associates have carried out the anlysis of fly ahs collected from municipal incinerators located in major urban centers in Japan, The Netherlands and Canada. They could detect at trace levels approximatlye 100 organic compounds which include PCDDs, PDDFs polychlorinated phenols, polychlorinated benzenes, polycyclic aromatic hydrocarbons, phthalate esters and aliphatic hydrocarbons. …..”The unexpected findings of PCDFs and PCDDs in the effluents from incinerators raise a few questions. Are PCDDs and relted compounds produced thermally in the incinerators. What are the mechanisms for the thermochemical formation of these undesired chloropollutants? What is the source of the chlorine?” There is still a problem with understanding this chemistry

These issues have not, at the present time, been resolved completely. Cf. Shibamo T et al., (2007) Rev Environ Contam Toxicol. 190: 1-41 who note that “more detailed investigation of many individual factors influencing dioxin formation is needed to find ways to reduce their formation in industrial and municipal incinerators”. It is suggested that the key substance to be considered in the evaluation of whether the incineration of municipal waste is a worthwhile activity, is the formation of HCB which controls the other less abundant dioxin and other products present. Another fairly recent review of HCB which includes information on health problems caused by this substances as well its formation in incinerators, by Barber J et al., (sponsored by Euro-Chlor available at http;//www.eurochlor.org/upload/documents/document 187.pdf (representing the chlor alkali industry which has historically been responsible for environmental pollution by similar products to those produced in municipal waste incinerators). This review is not overtly critical of the circumstances by which HCB and dioxins are formed as might be expected from an industrially-sponsored database, but nevertheless consideration of the data presented here should engender caution in inappropriate use of incineration of industrial and municipal waste as a convenient solution to the problem of disposal of industrial and municipal waste matter. (2b) US Dept of Health and Human Services, Agency for Toxic Substances and Disease Registry (Atlanta) Toxicological Profile for Hexachorobenzene United States Environmental Protection Agency at http://www.atsdr.cdc.gov/toxprofiles/tp90-c9.pdf (comprehensive list of references) Companion to http://www.atsdr.cdc.gov/toxprofiles/tp90.pdf This document attempts to provide a comprehensive review of toxicological data for HCB (but omits mention of CFS research); it is poorly edited but is a valuable source of information for suggesting future researches which may be of value for unravelling the cause of CFS and other illnesses. A wide species and individual difference in toxicological effects seems to be indicated. It should be noted that animal studies may be misleading for human health assessment. The low water solubility of HCB can result in port uptake from gavage. Data are lacking for trans-dermal uptake. More effective uptake seems to be achieved when administered orally with oil. Uptake, it should be noted is generally poor form airborne particulate matter, e.g. form occupational exposure. Blood and urine analysis may not provide adequate indications of HCB intoxication. There is also some evident of the occurrence of profound neurological symptoms (e.g. tremors) in subjects 30 years after ingestion of HCB. There was a high incidence of abortion and mortality in affected infants in the late 1950s Turkish intoxication incident where ingestion was estimated to be 0.7-2.9mg/kg/day for a 70kg person. The most sensitive target in adults for HCB intoxication and accumulation seems to be the ovaries.

A 90-day assay in adult female Cynomolgus monkeys showed ovarian toxicity occurred at 0.0001mg/kg/day (the lowest dose tested, cf. Bourque et al, 1995). HCB induced hyperparathyroidism and osteoproposid in male Fishcer 344 rate following gavage administration of 1mg.kg.day for 5 weeks ( Andrews et al., More general information on organochlorine pesticides is provided by http://www.cdc.gov/exposurereport/2nd/pdf/hexachlorobenzne.pdf and the internet posted document (at http://www.epa.gov/ncea/dioxin/part1/volume2/chap2.pdf “Mechanism of Formation of Dioxin-Like Compounds During Combustion of Organic Materials” [While this US government agency document was generally available on the internet it was provided with a request that it not be cited viz. each page is headed with the words “DRAFT-DO NOT QUOTE OR CITE”; this is presumably related to the possible legal status of this document in disputed origins of personal injury arising from the operation of municipal incinerators]. This report, however, gives a fairly comprehensive survey of the problem of generation of toxic substances during the incineration of wastes. The researches reviewed here seem, however, to have chose not to include older, but still highly relevant basic scientific research in the industrially sponsored researches in the field of “Structural Reorganization Throughout the Periodic Table” which defines from an academic scientific perspective the behaviour of molecules under pyrolysis conditions which might have seemed to be outwith the immediate field of incinerator science but, it can be suggested, is also to be highly relevant to the proper evaluation of that science. Cf. also “Bearing the Burden , Health Implications of Environmental Pollutants in Our Bodies A companion report to CDC’s 2003 Exposure Report ORGANOCHLORINE PESTICIDES http://www.cdc.gov/exposurereport/2nd/pdf/hexachlorobenzene Cf. Masuda Y (2003) Health effects of polychlorinated biphenyl and related compounds J Health Sci. 49 95) 333-336 And Taralunga M et al. (2004) Catalytic oxidation of chlorobenzene, a model compound for dioxin over Pt/zeolite catalyst Organohalogen Compounds 66: 1160-1166 (2c) Sharpe RM Irvine DS (2004) How strong is the link between environmental chemicals and adverse effects on human reproductive health? BMJ 328: 447-451 (3)

Toxicogenomics of HCB Intoxication Ezendam J Staedtler F Pennings J Vandebriel RJ Pieters R Harteman JH Vas JG (2004) Subchronic hexachlorobenzene exposure in brown Norway rats Environ Health Prespect Toxicogenomics 112 (7) 730-803 Autoimmune Activity of HCB (in the rat) Oral exposure to HCB showed stimulatory effects on spleen and lymph node weight and histological effects consistent with autoimmune activity. Increased IgM was observed in the Wistar rat. In the Lewis rat, HCB differentially modultaed autoimmunity Michielsen CCPPC, Van Voveren H Vos JG (1999) DATA (3b) Combined Effects of Mercury and HCB Intoxication Lecavalier PR et al. (1994) J Environ Sci Health B29 (5) 951-956 Human Industrial HCB Intoxication Data Queiros MLS Bincoletti C Perlington RCR Souza CR Toledo H (1997) Defective neutrophil function in workers occupationally exposed to hexachlorobenzene Hum Exp Toxicol 16: 322-326 Cf. [Same group] (1998) Immuunological levels in workers exposed to hexachlorobenzene And: Polymorphonuclear phagocytosis in workers occupationally exposed to hexachlorobenzene Immunopharmacol Immunotoxicol. 20 (3): 447-454 Ibid. 17: 172-175 Sala M Sunyer J herrero C To-Figueras J Grimault J (2001) Association between serum concentrations of hexachlorobenzene and polychlorobiphenyls with thyroid hormone and liver enzymes in a sample of the general population Occup Environ Med, 58 (3) 172-177 Prostate Cancer and HCB Intoxication Ralph et al. (2003) suggested that a possible correlation exists between HCB intoxication and steroid hormone regulation which could implicate high levels of HCB intoxication in the promotion of prostate cancer. Whereas low levels of HCB were associated with an augmented prostate activity, very high HCB intoxication levels were associated with overt carcinogenic damage. Disruption of Androgen Regulation Ralph JR Orgebin-Crist M-C Lareyre J-J Nelson CC (2003) Disruption of androgen regulation in the prostate by the environmental contaminant hexachlorobenzene Environ Health Perspect. 111: 461-433 Serum Organochorine Pesticide Intoxication and Risk of Testicular Germ Cell Carcinoma

Biggs ML et al., (2008) Serum organochlorine presticide residues and risk of testicular germ cell carcinoma: a population-based case-controlled study Cancer Epidem Biomarker Prev. 17 (8) 2012-2018 [This study did not support the hypothesis that exposure to organochlorine pesticides promotes testicular cancer]; Hardell L et al. (2004) Concentration of polychlorinated biphenyls in blood and the risk of testicular cancer Int J Androl. 27 (5) 282-290 [A previous study by this group had found a correlation between the occurrence of this disease and the POP (which included HCB) burden of case mothers (this cancer is believed to be induced in the foetus); a follow-on study identified PCB congeners as being of major concern for the induction of this disease]. Cf., Exposure to HCB During Pregnancy Induces Obesity in Children Smink A et al., (2008) Exposure to hexachlorobenzene during pregnancy increases the risk of overweight in children aged 6 years Acta Paed. 97 (10) 1465-1469 Dominant Role of HCB in Environmental Pollutants in Promoting the Global Obesity Epidemic P F Baillie-Hamilton (2002) Chemical toxins: a hypothesis to explain the global obesity epidemic J Alternative Comp Med. 8 (2): 185-192 [This wide-ranging review article discussed how mis-abstracting of scientific information could have obscured abundant data which suggests that low level intoxication by chemical molecules which are present as worldwide environmental pollutants is the origin the global epidemic of obesity as well as other illness. {Personal communication with the author confirmed that this review had identified eight previous reports which had implicated HCB intoxication as a possible cause of obesity. While other persistent environmental toxins including chlorinated pesticides may also be involved e.g. as synergistic factors, it can be rationally suggested on the basis of present evidence that the intoxication effects of HCB is the dominant factor which requires to be addressed}]. Cf., Association of HCB Intoxication with the Incidence of Diabetes Mellitus in Pregnant Women A Swedish study of chlorinated organic pollutants, in which the HCB, PCBs, dioxins and chlorinated insecticides using data which were corrected for regional variation, age and body weight indices of pregnant women showed up a highly significant correlation between HCB intoxication and the occurrence of diabetes. N.b. HCB intoxication alone and not intoxication by the other chlorinated environmental pollutants which were studied seems to be a putative major cause of diabetes. However a previous survey of a much later number of pregnant women in England had shown up a highly significant correlation between the intoxication by summed chlorinated biphenyls and diabetes. Glynn et al (2003) Environ Health Perspect. 222 (3) DATA,

[An age, body mass index and geographic corrected HCB level was strongly associated with diabetes mellitus a small survey of (7) pregnant women in Sweden. Diabetes was correlated with the presence of HCB but not with other chlorinated substances including PCBs and dioxins]. Association of HCB with the Incidence of Breast Cancer in Women DATA HCB in Th1 and Th2 Responses Different strains of rats respond differently to immunotoxic substances inclkudig the known immontoxin HCB which was used to probe Th1 and Th2 cytokine markers. Exposoure of Wistar rats to HCB had been known to alter Th1 IL-2 and IFNγ but not the supposed Th2 marker IL-4; a better Th2 marker, however, was suggested to be IL-10 which was decreased (both in production and expression) by HCB intoxication in Lewis rats. HCB had previously been reported to increase IgE levels in Brown Norway but not in (Th1 prone) Lewis rats, in which it aggravated experimental encephalomyelitis. Vanderbriel RJ et al (2000) Scand J Immunol 52 (5): 519-524 HCB in Serum and Urine of Airborne HCB Exposed Population To-Figueras J Sala M Otero R Barro C et al., (1997) Enviorn Health Perspect. 105 (1) 78-83 A study of the serum and urine from 100 subjects of a general population highly exposed to airborne HCB showed the presence of HCB in all serum samples (varying in content from 1.1-9.53ng/ml and its major known metabolite was present at 0.58-1.39µ g/24h (n.b. HCB is now indicated to be ubiquitously present in the global environment); a S-containing derivative of pentachlorobenzene was also detected, this apparently being the principal metabolite of HCB. Herrieoied et al. (1999) studies a groups of workers exposed chronically to HCB and reported no statistically relevant adverse health effects Nevertheless highly specific foetal development alteration (altered baby body lengths) were could be attributed to HCB intoxication of residents near Flix (Spain). A Japanese study failed to find any correlation between repeated miscarriages in women and their level of intoxication by multiple chlorinated organic substance pollutants which included HCB, A Canadian study, however, suggest that chlorinated organic pollutants were implicated in the promotion of female infertility. An epidemiological study of mid-aged and elderly women in Sweden showed up an unexpected significant correlation between HCB intoxication and the incidence of diabetes (however the population number was small). Herreroied C et al. (1999) Urinary porphyryia in urinary excretion in a human population highly exposed to hexachlorobenzene Arch Dermatol. 135: 400-404 Thomas DD Espey MG Vitek M Miranda KM Wink DA (2002) Protein nitration is mediated by heme and free metals through

Fenton-type chemistry: An alternative to the NO/.O2- reaction Proc Natl Acad Sci USA 99 (20): 12691-12696 (4) The comprehensive list of references given I the CDC Report (1) includes Iron Dyshomeostasis/ Lipid Peroxidation Allerman MA Kosater JF Wilson JHP et al.,(1985) The involvement of iron and lipid peroxidation in the pathogenesis of benzenenehexachloride induced porphyria [Also of relevance to HCB intoxication] Biochem Pharmacol. 34 (2): 161-166 Calcium Dyshomeostais Andrews JE et al., (1989) Impairment of calcium homeostasis by hexachlorobenzne exposure in Fischer 344 rats J Toxicol Environ Health 23: 311-320 Cf., Induction of Hyperparathyroidism and Osteosclerosis Andrews JE et al., (1990) Fundam Appl Toxicol. 12: 242-251 (5) Pentachlorophenol ex HCB Intoxication (N.b. pentachlorophenol is a major metabolite of HCB) To-Figueras J Salea M Otera R Barrot C et al. (1997) Enviorn Health Perspect. 105 (1); 78-83 Serum and urine from a general population highly exposed to airborne HCB showed HCB to be present at 1.0-9.5 ng/ml in serum etc., together with the major HCB metabolite pentachlorophenol (5a) Cf. De Meirlier K (2001) Flemish Parliament Discussion on ME/CFS Hearing of 5 March 2001 URL:http://jsp.vlaamsparlement.be/stukken/2000-2001/g740-1-BITC.pdf http://www.immunesupport,com/library.cfs/ID/3475/e/1/5 Dealt with pentachlorophenol intoxication in (a subgroup of) ME/CFS patients. This was one of seven suggested triggers. Others included heavy metals.)The possible role of household waste incinerators was broached) Immune dysregulation led to chronic infection which diminished the immune system of the host thereby augmenting chronic mycoplasmal and viral infections which were further believed to perturb the RNAseL antiviral pathway a side effect of which was the blockage of ion channels. K De Meirlier (Brussels) (considered one of the foremost medical experts on ME/CFS) was a principal invited speaker at a Royal Society of Edinburgh Symposium Held in Dundee (organised by Vance Spence) in 2003 in order to suggest novel therapeutic approaches to these illnesses. De Meirlier seemed to be especially concerned with promoting the drug Ampligen which is believed to correct post-viral infection dysregulation of an endogenous RNAse antiviral pathway.

(5b) RH Dunstan Group Identification of HCB Intoxication with CFS J Nutritional Environmental Medicine (Abingdon) June 1999 Suggested that low levels of exposure to organochlorines was associated with subtle yet significant changes in blood parameters which may not show up by standard procedures. Such parameters different between CFS and controls. Cf., http://implants.clic.net/tony/corner/A/0006.html Biochemcial and microbiochemical anomalies in chronic fatigue syndrome AHNF: 98 Conference: Dunstan http://www/ahmf.org/98dunstan2.html Chlorinated pesticides and chronic fatigue-related illness Cf. also Med J Aust 1995; 163: 294-297 A preliminary investigation of chlorinated hydrocarbons and chronic fatigue syndrome And Biochem Molec Med 58: DATA Bioaccumulated chlorinated hydrocarbons and red/white cell parameters Persons exposed to immune disrupter effects of chlorinated organic substances Could not be distinguished from CFS patients. This could even suggest that CFS could be a specific substance (most likely candidate HCB) induced disease. The long range transportation of HCB suggests that it has a uniquely general ability to cause a general population intoxication, which is due to HCB together with DDE and DDT which comprised more than 90% of the total amounts of the 24 chlorinated suspected immune disrupting organic agrochemicals. Dunstan et al. noted that HCB and the other chlorinated organics used as agrochemicals were much more accumulative in humans than were organophosphate agrochemicals. (6) Effect of Chitosan Oligosaccharides on 2,3,7,8-tetrachlrobenzene-p-dioxin – induced stress in mice Biol Pharm Bull. 25 (9) 1161-1164 Cf. also Massage Sauna Hydrocarbon Oil Ingestion These are amongst the number of alternative medical procedures which have been reported to show promise to diminish HCB in adipose tissue stores. Chitosan oligosaccharide treatment is believed, inter alia, to diminish oxidative damage to lipids caused by HCB Cf. Using chitosan to execute dioxin from human’s body http://lists.essential.org/dioxin-1/msg00820.html

Shon Y-H Nam Y-S (Dongguk University Korea) Effect of chitosan oligosaccharides on dioxin-induced CYP1A1 activity and lipid peroxidation http://www.chitosan.or.kr/intro1.01-6-3-3.html Shon YH et al., (2002) Biol Pharm Bull 25 (9): 1161-1164 Cf. also Kim TY Jeong HG et al., (2004) Induction of iNOS and pro-inflammatory cytokines by o,pI-DDT in macrophages Toxicol Lett 146 (3): 261-269 (6a) Guar Gum Nakashima Y et al., (1998) Masking of guar gum induced acceleration of hexachlorobenzene excretion by its rapid excretion through lactation in adult female rats J Agric Food Chem. 46 (6) 2241-2247 (6b) Fish Oil Umegaki et al., (1998) Feeding fish oil to rats accelerates the metabolism of hexachlorobenzene J Nutr Sci Vitaminol. 44: 301-311 Suppression of γ IFN by HCB Daniel V et al. (2001) Environ Health Perspec 109 (2): 173-178 [This work reported the association of blood levels of PCB, HCH and HCB With numbers of lymphocyte subpopulations, etc.] HCB Stimulation of Liver Heme Oxygenase (HO-1) (an antioxidant heat shock protein) Stonaard MD et al. (1998) Arch Toxicol 72 (6) 355-361 [Cf Amy Bishop Research http://www.uah.edu/colleges/science /biology/amy/amy.htm Induced adaptive resisitance (IAR) to NO in the CNS. IAR is dependent on the hemem metabolising enzyme heme oxygenase-1 (HO-1) which limits protein nitration associated with neuronal cell death] Malic Acid Gene Transcription (Krebs Cycle Shunt) Loazia-Perez AI Seisdedos M-T Kleiman de Pisarev DL Sancovich HGA Randi AS Feerramole de Sancovich AM Santisteban P (1999) Hexachlorobenzene, a dioxin-type compound, increases malic enzyme gene transcription through a mechanism involving the thyroid response element Endocrinology 140: 4142-4151 [This study was conducted with the rat]

Tryptophan Metabolism Lambian EB et al. DATA α− Lipoic Acid Vilas GI et al. (1999) Effect of α lipoic acid on ehxachlorobenzen prophyria Biocnem Med Biol Int. 47 (5): 815-823 γ -Glutamyl Transferase Adjarov D et al., (1982) Toxicology 23: 73-77 [This provides a sensitive marker for experimental hexachlorobenzene intoxication] Resistance to Thyroid Tolerance. Hypothesis of FM etc. Garnace RL Bredding PC (2000) A metabolic basis fro fibromyalgia and realtaed disorders, the possible role of resistance to thyroid tolerance Med Hypoth. 61 (2) 132-139 Serotonin Turnover and HCB DATA

Potential Therapeutics Vegan Diet and FM Aartinum K Lammi K Hypen M Nenonen M Hauina O Kauma M (2000) Vegan diet alleviates fibromyalgia symptoms Scand J Rheumatol. 29 (5): 308-313 Cf also (Kaartinen et al., (2000) Addendum Adrenal bioactivation and toxicity of 3-MeSO2-DDE, o.pl-DDD and DMBA Lindhe O (2001) Acta Universitatis Upsaliensis Dissertations from the Faculty of Science & Technology 695 49pp investigated in tissue slice culture Cf. Suppression of sulfotransferase activity by 2,3,7,8 tetrachlorodibenzo-pdioxin Dusanmer Z et al. (2001) Drug Metab Deposition 29: 1130-1135 Hyaluronan in Blood Serum (e.g. in FM)) Yaron I Buskela D Shirazi I Neuman L Elkayam O Paran D Yar M (1997) J Rheumatol. 24 (11): 2221-2224 Cf.. ibid., (1999) 26 (9) 2063 and (2002) 28 (11) 2561

HCB Contamination of Food and Dietary Treatment for Putative HCB-Intoxication Related Diseases. Since HCB is highly resistance to degradation in the environment it becomes an ubiquitous contaminant of milk, meat and fish and human adiopose tissue. The human daily intake of HCB is estimated as 0.13µ g/person. Obviously, avoidance of HCB-contaminated foods needs to be considered as a strategy to counter HCB-led illnesses. This idea is supported by the beneficial effect of a meat, fish and milk-free diet which seemed to produce a marked diminution of the symptoms of a CFS-related illness (fibromyalgia (FM)) in treated compared with control groups. Are Thyroid Disrupting Agents (e.g.HCB) Responsible for Fibromyalgia (FM)? The serum of women with fibromyalgia showed a large increase in serum hyaluronan fragments. This finding was suggested to be a useful diagnostic test for FM. (Yaron et al. 1997; Burkhuizer & Bennett 1999). These findings have prompted the thyroiditis hypothesis of FM and ME/CFS. (Cf. Garnace & Breeding 2000). Disagreement however exists between the possible effect of an additional cofactor apparently present in an Israeli group which seemed to be absent in a Danish group later studied which seemed not to confirm the hyaluronan data obtained from an Israeli group. It can be rationally suggested that the primary insult of HCB intoxication and resultant pro-oxidant stress leads to hyaluronan fragmentation as well as thyroid dysregulation (such dysregulation had been previously empirically associated with the amounts of hyaluronan in blood serum). Hyaluronan was also found to be increased but by a much lesser amount in the serum of patients with rheumatoid arthritis. α− Lipoic acid dietary supplementation has also been reported to suppress HCBinduced porphyria (5). (Other) antioxidant supplementation seems warranted to inhibit the general pro-oxidant effects of HCB intoxication. It has been found that the antioxidant quercetin, ingested in the pure chemical form, can give relief from the symptoms of CFS. (N.b. this antioxidant is also present in fruit e.g. in apple skins the ingestion of which may be of value for combating HCB intoxication). Dundee ME/CFS Research Group Findings Preliminary studies conducted on ME/CFS patients by Gail Kenendy and Vance Spence (cf., Spence (2004) and also discussed in the 2003 Dundee Royal Society Workshop on ME) apparently showed up difference between Gulf War Syndrome, ME, ME/CFS and organophosphate intoxication by using acetylcholine vasoconstrictor sensing procedures which were thought to function via the inhibition of acetylcholinesterase and related nitric oxide functions. It is suggested that these techniques could usefully be used to test the effect of HCB intoxication. Since heparan sulphate biochemical control systems are highly implicated in function and protection of the vasculature it is a reasonable hypothesis that the changes apparent in these organs in patients with ME-like diseases involve alteration in heparan sulphate microstructure (e.g. consequent on an elevation of oxidant/nitrant damaging moieties is a result of pro-oxidant intoxication arising from HCB (together with other chlorinated organic pollutants). A further extension of this hypothesis suggests that HCB may impacts on the heparan sulphate system via its action on thyroid factors which are known to be involve in

transfer of inorganic sulphate needed for sulphation of heparan sulphate in the Golgi apparatus. The heparan sulphate mimetic chitosan sulpahte has been suggested to provide protection against HCB intoxication perhaps by mimicking some of the endogenous anionic polysaccharide activities. Formation of HCB from Lindane is known to occur in vivo. HCB has also been positively associated with CD4:CD8 ratio and neutrophil count in CFS patients. The existence of HCB intoxication in CFS-like illnesses could also explain the greater incidence of bacterial infection caused by reduced phagocytosis activities of HCB intoxicated macrophages and neutrophils. Is HCB a Pre-Dioxin? Dioxins, especially 2,3,7,8 TCDD are considerable more toxic on a molar basis than is HCB, but it seems to be biochemically possible that HCB may also be transformed in vivo into 2,3,7,8TCDD/TDDF. Further research is required to establish the relative toxicity of HCB and its metabolites and to further probe the mechanism by which HCB seem greatly enhance, by an apparent synergistic mechanism, some of the toxic actions of 2,3,7,8TCDD. Other reports have indicated that synergism exists between HCB intoxication and that of mercury as well as of nicotine. This could also be of possible relevance to the aetiology of ME/CFS, and require further research as does the indication the HCB is able to increase the severity of bacterial infection and membrane damage (e.g. caused by staphylococcus toxin).