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The insidious and deadly nature of mercury's organometallic compounds is informed by two large scale poisonings due to industrial mercury pollution that occurred decades ago in Minamata and Niigata, Japan. The present study examined chemical speciation for both mercury and selenium in a historic umbilical cord sample from a child born to a mother who lived near the Agano River in Niigata. The mother had experienced mercury exposure leading to more than 50 ppm mercury measured in her hair and was symptomatic 9 years prior to the birth. We sought to determine the mercury and selenium speciation in the child's cord using Hg Lα1 and Se Kα1 high-energy resolution fluorescence detected X-ray absorption spectroscopy, the chemical speciation of mercury was found to be predominantly organometallic and coordinated to a thiolate. The selenium was found to be primarily in an organic form and at levels higher than those of mercury, with no evidence of mercury-selenium chemical species. Our results are consistent with mercury exposure at Niigata being due to exposure to organometallic mercury species., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Graham George reports financial support was provided by Natural Sciences and Engineering Research Council of Canada. Ingrid Pickering reports financial support was provided by Canada Research Chairs. Ingrid Pickering reports financial support was provided by Natural Sciences and Engineering Research Council of Canada. Graham George reports financial support was provided by Canada Research Chairs. Graham George reports financial support was provided by New Frontiers in Research Fund. Ingrid Pickering reports financial support was provided by Canada Foundation For Innovation. Gary Myers reports financial support was provided by National Institute of Environmental Health Sciences., (Copyright © 2023. Published by Elsevier B.V.)

In situ methods are valuable in all fields of research. In toxicology, the importance of dose is well known, elevating the need for in situ techniques to measure levels of toxicants and their byproducts in precise anatomically identifiable locations. More recently, additional emphasis has been placed on the value of techniques which can detect chemical form or speciation, which is equally important in the toxicology of a chemical compound. Many important but conventional methods risk losing valuable information due to extractions, digestions, or the general reliance on mobile phases. Few analytical tools possess the power and diversity of X-ray methods as in-situ methods. Here we present an overview, intended for toxicologists and pathologists, of a variety of synchrotron X-ray methods for determining in situ chemical form and distribution of heavier elements. The versatility and range of these synchrotron techniques, which are both established and emerging, is demonstrated in the context of the study of neurotoxicology of mercury, a global pollutant with the ability to harm both human health and the environment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Mercury is ubiquitous in the environment, with rising levels due to pollution and climate change being a current global concern. Many mercury compounds are notorious for their toxicity, with the potential of organometallic mercury compounds for devastating effects on the structures and functions of the central nervous system being of particular concern. Chronic exposure of human populations to low levels of methylmercury compounds occurs through consumption of fish and other seafood, although the health consequences, if any, from this exposure remain controversial. We have used high energy resolution fluorescence detected X-ray absorption spectroscopy to determine the speciation of mercury and selenium in human brain tissue. We show that the molecular fate of mercury differs dramatically between individuals who suffered acute organometallic mercury exposure (poisoning) and individuals with chronic low-level exposure from a diet rich in marine fish. For long-term low-level methylmercury exposure from fish consumption, mercury speciation in brain tissue shows methylmercury coordinated to an aliphatic thiolate, resembling the coordination environment observed in marine fish. In marked contrast, for short-term high-level exposure, we observe the presence of biologically less available mercuric selenide deposits, confirmed by X-ray fluorescence imaging, as well as mercury(II)-bis-thiolate complexes, which may be signatures of severe poisoning in humans. These differences between low-level and high-level exposures challenge the relevance of studies involving acute exposure as a proxy for low-level chronic exposure.

Hydroquinone (HQ) exposure is common as it is a natural component of plant-based foods and is used in some fingernail polishes, hair dyes, and skin lighteners. Industrially it is used as an antioxidant, polymerization inhibitor, and reducing agent. The current study was undertaken to determine whether HQ may cause DNA damage in an in vivo comet assay in F344 rats. DNA strand breaks were assessed in the duodenum as a direct tissue contact site, the testes, and the liver and kidneys, which were tumor sites in bioassays. Rats were exposed to HQ by gavage at 0, 105, 210, or 420 mg/kg/day. At all dose levels, mean % tail intensity and tail moment values for all tissues in animals given HQ were similar to the control. There were no statistically significant increases in tail intensity in any tissue following HQ treatment of male and female rat and data for all animals fell within the available historical control ranges for each tissue. There was no evidence of induction of DNA damage in cells isolated from duodenum, kidney or liver of male and female rats or in the testes of male rats following exposure to HQ at a dose levels up to 420 mg/kg/day, which caused acute renal necrosis., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: John O’Donoghue is a consultant toxicologist who prepared this manuscript for the Hydroquinone Consortium. Carol Beevers is a consultant genetic toxicologist, who assisted in the preparation of this manuscript and during her previous employment at Covance Laboratories Ltd, Harrogate, UK, was the study director responsible for the conduct of the GLP comet assay described in this manuscript. Annie Buard is an employee of Solvay (Regulatory Toxicologist in the HSE / Product Regulatory Affairs - Product Safety Pole), which is a member company of the Hydroquinone Consortium., (© 2021 Published by Elsevier B.V.)

Background: Human exposure to mercury (Hg) is widespread and both organic and inorganic Hg are routinely found in the human brain. Millions of people are exposed to methyl Hg (MeHg) due to the consumption of fish and to inorganic Hg from dental amalgams, small scale gold mining operations, use of Hg containing products, or their occupations. Neuropathology information associated with exposures to different species of Hg is primarily based on case reports of single individuals or collections of case studies involving a single species of Hg at toxic exposure levels such as occurred in Japan and Iraq., Methods/results: This study brings together information on the neuropathological findings and deposition of Hg in the central nervous system of people exposed to different species of Hg at varying concentrations. The low dose exposures were lifetime exposures while the high dose exposures were generally acute or short term by different exposure routes with survival lasting various lengths of time. Total and inorganic Hg deposits were identified in formalin-fixed, paraffin embedded tissues from both low and high exposure Hg cases. Low concentration exposures were studied in adult brains from Rochester, New York (n = 4) and the Republic of Seychelles (n = 17). Rochester specimens had mean total Hg concentrations of 16-18 ppb in the calcarine, rolandic, and cerebellar cortices. Inorganic Hg averaged between 5-6 ppb or 30-37% for the cerebral and cerebellar cortices of the Rochester subjects. Total Hg was approximately 10-fold higher in specimens from Seychelles, where consumption of ocean fish is high and consequently results in exposure to MeHg. The predominant Hg species was MeHg in both the Rochester and Seychelles brain specimens. Histologically, cerebral and cerebellar cortices from Rochester and Seychelles specimens were indistinguishable. High concentration exposures were studied in brains from four adults who were autopsied at variable time periods after exposure to organic Hg (methyl or dimethyl) or inorganic Hg (inhaled vapor or intravenous injection of metallic Hg). In contrast to the Seychellois adults, these individuals had acute or subacute exposures to lethal or significantly higher concentrations. The pattern of Hg deposition differed between subjects with high organic Hg exposure and high inorganic Hg exposure. In the organic Hg cases, glia (astrocytes and microglia) and endothelial cells accumulated more Hg than neurons and there were minimal Hg deposits in cerebellar granule and Purkinje cells, anterior horn motor neurons, and neocortical pyramidal neurons. In the inorganic Hg cases, Hg was seen predominantly in neurons, vascular walls, brainstem, and cerebellar and cerebral deep gray nuclei. The presence of inorganic Hg in neural and neural supporting cells in the four high exposure Hg cases was not closely correlated with cellular pathology; particularly in the inorganic Hg cases., Conclusions: Different Hg species are associated with differing neuropathological patterns. No neuropathological abnormalities were present in the brains of either Rochester or Seychelles residents despite substantial differences in dietary MeHg exposure. Increasing concentrations of inorganic Hg were present in the brain of relatively low exposure subjects with increasing age., Competing Interests: Declaration of Competing Interest The author declare that they have no known competing financial interests or personal relationship that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Industrial release of mercury into the local Minamata environment with consequent poisoning of local communities through contaminated fish and shellfish consumption is considered the classic case of environmental mercury poisoning. However, the mercury species in the factory effluent has proved controversial, originally suggested as inorganic, and more recently as methylmercury species. We used newly available methods to re-examine the cerebellum of historic Cat 717, which was fed factory effluent mixed with food to confirm the source. Synchrotron high-energy-resolution fluorescence detection-X-ray absorption spectroscopy revealed sulfur-bound organometallic mercury with a minor β-HgS phase. Density functional theory indicated energetic preference for α-mercuri-acetaldehyde as a waste product of aldehyde production. The consequences of this alternative species in the "classic" mercury poisoning should be re-evaluated.

Human exposure to potentially neurotoxic methylmercury species is a public-health concern for many populations worldwide. Both fish and whale are known to contain varying amounts of methylmercury species. However studies of populations that consume large quantities of fish or whale have provided no clear consensus as to the extent of the risk. The toxicological profile of an element depends strongly on its chemical form. We have used X-ray absorption spectroscopy to investigate the comparative chemical forms of mercury and selenium in fish and whale skeletal muscle. The predominant chemical form of mercury in whale is found to closely resemble that found in fish. In the samples of skeletal muscle studied, no involvement of selenium in coordination of mercury is indicated in either whale or fish, with no significant inorganic HgSe or HgS type phases being detected. The selenium speciation in fish and whale shows that similar chemical types are present in each, but in significantly different proportions. Our results suggest that for equal amounts of Hg in skeletal muscle, the direct detrimental effects arising from the mercury content from consuming skeletal muscle from whale and fish should be similar if the effects of interactions with other components in the meat are not considered.

Methylmercury is among the most potentially toxic species to which human populations are exposed, both at high levels through poisonings and at lower levels through consumption of fish and other seafood. However, the molecular mechanisms of methylmercury toxicity in humans remain poorly understood. We used synchrotron X-ray absorption spectroscopy (XAS) to study mercury chemical forms in human brain tissue. Individuals poisoned with high levels of methylmercury species showed elevated cortical selenium with significant proportions of nanoparticulate mercuric selenide plus some inorganic mercury and methylmercury bound to organic sulfur. Individuals with a lifetime of high fish consumption showed much lower levels of mercuric selenide and methylmercury cysteineate. Mercury exposure did not perturb organic selenium levels. These results elucidate a key detoxification pathway in the central nervous system and provide new insights into the appropriate methods for biological monitoring.

Hydroquinone (HQ) is a common water-soluble constituent of foods, an ingredient in skin lightening preparations, a photographic developer, and an antioxidant used in the preparation of industrial polymers. In this series of studies, aqueous solutions of HQ were given by gavage to male and female Sprague-Dawley rats to determine the acutely lethal dose, the clinical signs of behavioral toxicity associated with doses at or near a dose causing mortality, and the effects of the administration of dose levels resulting in acutely observable behavioral effects when administered 5 days/week for 13 weeks. The acute dermal toxicity of HQ in rabbits was also determined. For the acute oral toxicity study, groups of five male and five female rats were administered single oral doses of 375, 345, 315, or 285 mg/kg. At all dose levels, animals exhibited minor to moderate tremors and minor convulsions within the first hour after dosing. The acute oral LD50 value for both sexes combined was >375 mg/kg. Dermal application of 2000 mg/kg HQ to rabbits under an occlusive wrap for 24 h did not result in neurobehavioral effects or mortality. Subchronic exposure was accomplished by administration of doses of 200, 64, 20, or 0 mg/kg/day of HQ in water to groups of male and female rats study (10/sex/group). A functional observational battery (FOB) was used to detect neurobehavioral effects prior to HQ exposure and postexposure at 1, 6, and 24 h and 7, 14, 30, 60, and 91 days. Daily clinical observations were also recorded for each animal. Doses of 200 or 64 mg/kg HQ resulted in acutely observable behavioral effects including tremors and reduced activity. Tremors occurred within one hour of dosing and resolved by the 6-h examination. Brain weights were not altered by HQ administration, but mean terminal body weight was reduced approximately 7% for the 200 mg/kg males. Neuropathologic examination of the CNS and PNS, including special stains for myelin and axonal process, did not reveal any morphologic lesions associated with HQ administration or secondary to repetitive CNS stimulation by HQ. The nephrotoxic effects observed in Fischer 344 rats after HQ exposure was not observed in this study with Sprague-Dawley rats. Oral doses of >or=64/mg/kg HQ resulted in acute neurobehavioral effects indicative of CNS stimulation; however, subchronic exposure to dose levels that produced repetitive CNS stimulation by HQ did not result in an exacerbation of acute stimulatory effects over time or morphologic changes in the CNS or PNS or nephrotoxicity.

Hydroquinone (HQ) has been used since the 1950s in commercially available over-the-counter skin lightener products and since the 1960s as a commercially available medical product. It is also used in cosmetic products such as hair dyes and products for coating finger nails. Beginning in 2001, HQ is no longer authorized for use in cosmetic skin lightening formulations in European Union countries, although products containing arbutin, an analogue of HQ, and botanicals, including plants that naturally contain HQ and arbutin, continue to remain available in European countries. The potential toxicity of HQ is dependent on the route of exposure, and toxicity in rodents is highly sex-, species-, and strain-specific. Subchronic and chronic toxicity in experimental animals is primarily limited to nephrotoxicity in male F-344 rats. Dermal toxicity studies, even those conducted in the sensitive male F-344 rat, are essentially devoid of systemic toxicity. Developmental and reproductive toxicity studies with HQ in rats and rabbits have not demonstrated significant effects. Cancer bioassay data for HQ demonstrate limited effects and are not sufficient to classify HQ for human carcinogenicity. Epidemiology and occupational studies of workers with extensive exposure to HQ have not reported any evidence of adverse systemic health effects or carcinogenicity. A risk-benefit approach is recommended for assessing the available data for HQ, arbutin, and other materials in use as, or proposed for use as, skin lighteners to provide optimal therapeutic benefits to patients with pigmentary changes of the skin.

Given the emerging interest among researchers, practitioners, and policymakers in youth participation, it is important to examine and assess carefully the promise and challenges of youth engagement.

Methyl iso-butyl ketone (MiBK), a commercial solvent, was selected by the US Environmental Protection Agency (US EPA) for testing under the Multi-Substance Rule for the Testing of Neurotoxicity (US EPA, 1993) using schedule-controlled operant behavior (SCOB) to determine if subchronic exposure to MiBK vapor had the potential to alter behavior as an indicator of neurotoxicity. Food-restricted and ad libitum-fed Sprague-Dawley male rats were exposed to 0, 250, 750, or 1500 ppm MiBK for 6 h/day, 5 d/wk for 13 weeks. SCOB testing of food-restricted animals, using a multiple fixed ratio (FR)/fixed interval (FI) schedule (FR20:FI120), was conducted prior to each exposure to maintain the operant behavior; the data from Weeks -1, 4, 8, and 13 were evaluated for evidence of neurotoxicity. SCOB testing was also evaluated for two weeks following the cessation of exposures. Ad libitum-fed animals were included to assess systemic effects using routine indicators such as changes in body weight, food consumption, and organ weight. No significant differences were seen in fixed-ratio run rate, FR pause duration, fixed-interval response rate, and index of curvature values at any concentration. Animals exposed to 750 and 1500 ppm MiBK exhibited clinical signs associated with transient reduced activity levels, but only during exposure. No signs of reduced activity were observed immediately after exposure for either group. No other treatment-related abnormalities were observed during exposure. Food-restricted animals did not demonstrate any increased or decreased sensitivity to the CNS depressive effects of MiBK relative to the ad libitum-fed animals. No treatment-related body weight differences were observed within either the food-restricted groups or the ad libitum-fed groups, although body weights of the former were clearly depressed compared with those of the latter. Relative and absolute liver, and relative kidney weights were significantly greater for the 750 and 1500 ppm ad libitum-fed animals. No differences in kidney weight were observed for food-restricted animals, but absolute and/or relative liver weights were significantly higher for all the treated food-restricted groups. The results of this study indicate that repetitive exposures to high concentrations of MiBK vapors do not result in adverse effects on operant behavior in the rat.

n-Butyl acetate, a common industrial solvent, was selected by the US EPA as a chemical of concern for neurotoxicity as part of the Multisubstance Rule for the Testing of Neurotoxicity. The neurotoxic potential of n-butyl acetate was investigated in Sprague-Dawley rats using a functional observational battery, motor activity, neurohistopathology, and schedule-controlled operant behavior (SCOB) as indicators of neurotoxicity. Animals were exposed to concentrations of 0, 500, 1500, or 3000 ppm of n-butyl acetate for 6 hours per day for 65 exposures over 14 weeks. Functional observational battery and motor activity values for ad libitum-fed male and female rats were measured during Weeks -1, 4, 8, and 13. SCOB testing of food-restricted animals, using a multiple fixed ratio/fixed interval schedule, was conducted daily prior to each exposure to maintain the operant behavior; the data from Weeks -1, 4, 8, and 13 were evaluated for evidence of neurotoxicity. Transient signs of sedation and hypoactivity were observed only during exposure to the 1500 and 3000 ppm concentrations. The only signs of systemic toxicity were reduced body weights for the 3000 ppm ad libitum-fed groups and occasionally for the female 1500 ppm ad libitum-fed group. No evidence of neurotoxicity was seen during the functional observational battery examinations. Motor activity for the 3000 ppm male group was significantly (p < or = 0.05) higher than for the control group only during Week 4. No significant differences were observed among groups for Weeks 8 and 13. No significant differences in motor activity values were observed for female rats. No significant differences were seen in operant behavior at any test vapor concentration. Microscopic evaluations of sections from the brain, spinal cord (cervical and lumbar regions), dorsal and ventral spinal roots, dorsal root ganglia, sciatic nerve, and tibial nerve of animals in the control and 3000 ppm groups did not indicate any treatment-related effects. In conclusion, there was no evidence of cumulative neurotoxicity based on the functional observational battery, motor activity, neurohistopathology, and schedule-controlled operant behavior endpoints. The data presented here are relevant to the neurotoxicity risk assessment of n-butanol due to the rapid hydrolysis of n-butyl acetate in vivo.

Hydroquinone (HQ) is used in over-the-counter formulations of skin-lightening creams sold in the United States and European Union. HQ was introduced into these formulations to provide a safe and effective alternative to mercury and other less effective ingredients. Recent studies involving subchronic oral exposure of male F344 rats to HQ have shown nephrotoxicity and renal tubule cell proliferation (English et al., 1994), while chronic exposures of male F344 rats were reported to cause renal cell adenomas (NTP, 1989). Previous subchronic dermal toxicity studies (CTFA, 1986; NTP, 1989) with HQ failed to detect nephrotoxicity; however, these studies were not specifically designed to assess renal structure and function. More sensitive endpoints were used in the present subchronic study to address concerns over potential toxicity from repeated dermal exposure to HQ. Male and female F344 rats were given topical applications with 0, 2.0, 3.5, or 5.0% HQ in an oil-in-water emulsion cream for 13 wk (5 days/wk). Body weights, feed consumption and water consumption were monitored, and animals were observed for clinical signs of toxicity and dermal irritation. Blood taken at termination was analysed for haematological and clinical chemistry effects. Erythema, which abated when exposure stopped, was the only dermatological effect seen at the HQ-cream application sites. Cell proliferation in the kidneys was evaluated after 3, 6 and 13 wk of treatment using bromodeoxyuridine (BrdU) labelling, but no changes indicative of sustained cell proliferation were seen. The renal histopathological lesions noted after oral exposure to HQ were not present after dermal exposure. Thus, topical exposure to HQ does not result in the renal toxicity observed in previous studies with F344 rats given HQ orally.

The proposed Neurotoxicity Risk Assessment Guidelines (U.S. EPA, 1995c Fed. Reg. 60(192), 52032-52056) of the U.S. Environmental Protection Agency (EPA) were the subject of a workshop at the 1997 Meeting of the Society of Toxicology. The workshop considered the role of guidelines in the risk assessment process, the primary features, scientific basis, and implications of the guidelines for EPA program offices, as well as for industrial neurotoxicologists from the perspectives of both pesticides and toxic substances regulation. The U.S. National Academy of Sciences (NAS, 1983, Risk Assessment in the Federal Government: Managing the Process) established a framework for distinguishing risk management from risk assessment, the latter being the result of integrating hazard identification, hazard characterization, and exposure assessment data. The guidelines are intended to establish operating principles that will be used when examining data in a risk assessment context. The proposed neurotoxicity risk assessment guidelines provide a conceptual framework for deciding whether or not a chemically induced effect can be considered to be evidence of neurotoxicity. Topics in the proposed guidelines include structural and functional effects, dose-response and -duration considerations, and relationships between effects. Among the issues that must be considered are the multiplicity of chemical effects, the levels of biological organization in the nervous system, and the tests, measurements, and protocols used. Judgment of the adversity of an effect depends heavily on the amount and types of data available. The attribution of a chemically induced effect to an action on the nervous system depends on several factors such as the quality of the study, the nature of the outcome, dose-response and time-response relationships, and the possible involvement of nonneural factors. The guidelines will also serve as a reference for those conducting neurotoxicity testing, as well as establish a consistent approach to neurotoxicity risk assessment by regulators. Extending this approach through international harmonization would be advantageous to the development of products for a worldwide market. Thus, both risk assessors and regulated industries have a large stake in the guidelines to provide a framework that will lead to accurate risk assessment decisions.

The fundamental structures and functions of the nervous systems of animals and humans are conserved in many ways across species. These similarities provide a basis for developing common neurologic examinations for a number of species of animals and also provide a basis for developing risk assessments across species for neurologic end points. The neurologic examination requires no expensive equipment and can be conducted in the field or wherever impaired animals are identified. The proper conduct of neurologic examinations in animals assumes that the examiner has a fundamental understanding of the normal structure and function of the nervous system as well as knowledge about the spontaneous disease background of the species being studied.

Mortality in a 1942-1990 cohort of 858 men and 21 women employed in the manufacture and use of hydroquinone was evaluated through 1991. Average exposure concentrations, 1949-1990, ranged from 0.1 to 6.0 mg/m3 for hydroquinone dust and from less than 0.1 to 0.3 for quinone vapor (estimated 8-h time-weighted averages). Compared with general population and occupational referents, there were statistically significant deficits in total mortality and deaths due to cancer. No significant excesses were observed for such hypothesized causes as kidney cancer [2 observed vs 1.3 expected (both control groups), P approximately 0.39], liver cancer (0 vs 0.8, 1.3), and leukemia (0 vs 2.3, 2.7). Dose-response analyses of selected causes of death, including renal carcinoma, demonstrated no statistically significant heterogeneities or linear trends according to estimated career hydroquinone exposure (mg/m3-years) or time from first exposure.

Oral administration of hydroquinone (HQ) over 2 years to male Fischer 344 (F344) rats results in a dose-related nephropathy and an increase in the incidence of renal tubule adenomas. Female F344 rats, B6C3F1 mice, and Sprague-Dawley (SD) rats are resistant to the chronic renal toxicity of HQ, and nephrotoxicity was not seen in dogs or humans following subchronic exposure. To better characterize the early development of renal toxicity in rats, cell proliferation was quantitated within the proximal (P1, P2, and P3) and distal tubule segments of the kidney in rats given 0, 2.5, 25, or 50 mg/kg HQ by gavage. Male and female F344 rats were treated for 1, 3, or 6 weeks, and male SD rats were treated for 6 weeks. Cell proliferation was quantitated by incorporation of bromodeoxyuridine, detected immunohistochemically, into newly synthesized DNA. At 6 weeks, renal cell proliferation was increased over vehicle-controls in male F344 rats dosed at 50 mg/kg. Significant elevations (p < 0.001) occurred in the P1 segments (87%) and in the P2 segments (50%) but the elevation in the P3 segment (34%) was not statistically significant. Urinalyses revealed increases in the rate of excretion of enzymes indicative of proximal tubular damage. Histopathologic evaluation of the kidneys was consistent with a dose-related tubular degeneration in the male F344 rat. No chemical-related effects were observed in the kidneys of female F344 and male SD rats. These data parallel the findings of sex- and strain-specific kidney adenomas in the 2-year bioassays, and suggest that chemically induced cell proliferation secondary to toxicity may be important in the pathogenesis of benign renal tumors in male F344 rats treated with HQ.

Oral administration of hydroquinone (HQ) to male Fischer 344 (F344) rats results in dose-related kidney toxicity beginning with mild enzymuria by 1 week, significant cell proliferation by 6 weeks, and nephropathy and an increase in the incidence of renal tubule adenomas after 2 years. Female F344 rats, B6C3F1 mice, Sprague-Dawley rats, dogs, and humans are resistant to the renal toxicity of HQ associated with repeated exposure. To determine the potential of HQ to induce covalent DNA adducts in the kidney, male and female F344 rats were given 0, 2.5, 25, or 50 mg/kg HQ by gavage for 6 weeks, and nuclear DNA isolated from kidneys was analyzed by the 32P-postlabeling assay. At 50 mg/kg, males, but not females, showed an increase in the rate of excretion of N-acetyl-beta-D-glucosaminidase, indicative of proximal tubular damage. Analysis of nuclear DNA preparations by the postlabeling assay showed that HQ does not produce covalent DNA adducts in the kidneys of male and female rats. The assay's lower limit of detection is 1 adduct in 10(9) to 10(10) DNA nucleotides. No treatment-related increases in background radioactivity levels on the chromatograms were seen at locations corresponding to the major in vitro adducts of HQ and p-benzoquinone. HQ treatment, however, resulted in the reduction of the levels of certain endogenous adducts (I-compounds), the biological significance of which is unknown.(ABSTRACT TRUNCATED AT 250 WORDS)

Hairless guinea pigs were born in a closed colony of Hartley guinea pigs. At birth, hairless guinea pigs were smaller than littermates, had wrinkled skin and stunted vibrissae. The most striking abnormality histologically was distension of the upper portion of the pilary canal with thickening of the epidermis. Hairlessness resulted from production of abnormal hair shafts. Preliminary work indicated that in addition to hairlessness, the guinea pigs were athymic or hypothymic. Initial immunologic studies showed that the guinea pigs also were agammaglobulinemic. Animals that survived the weaning period died of infections that are usually associated with immunodeficiencies, such as systemic cytomegalovirus, systemic balantidiasis and Pneumocystis carinii pneumonia.

Groups of five male and five female rats were fed diets containing from 0% to 2.5% di(2-ethylhexyl)terephthalate (DEHT) or 1.2% di(2-ethylhexyl)phthalate (DEHP) for 21 days. Feed consumption and body weight gains were collected and, at study termination, animals were examined for alterations in body weight, differences in serum lipids, changes in the activities of certain enzymes associated with fat metabolism, and proliferation of hepatic peroxisomes. Feed consumption and weight gain were greatly decreased in DEHT-fed animals only at 2.5%. No biologically significant alterations in absolute liver weight occurred with DEHT. Relative liver weights were increased at 2.5% in both sexes and at 1.0% and 1.2% in females. The alterations were due wholly to decreased terminal body weights. Serum triglyceride and cholesterol levels were not found useful in interpreting the effects of DEHT. Cyanide-insensitive palmitoyl CoA oxidation and lauric acid 11- and 12-hydroxylation were increased in animals consuming 2.5%, but no lower levels of DEHT. Induction of hepatic peroxisomes did not occur at 1.2% DEHT. Interpretation of minimal peroxisomal effects with 2.5% DEHT was confounded by reduced feed consumption. Slight decreases in weight gain occurred in males consuming the 1.2% DEHP diet, but differences were minor relative to effects observed at 2.5% DEHT. Results with DEHP contrasted with those obtained with DEHT. Absolute and relative liver weights, activities of enzymes of lipid metabolism, and peroxisome content were all significantly increased at 1.2% DEHP. Reduction of feed intake was implicated in the effects observed at 2.5% DEHT, since the amount of DEHT consumed by 2.5% animals was only 1.4 times as much as by 1.2% animals. A possible explanation for the observed differences between DEHP and DEHT was related to the results of a metabolic fate study on DEHT. Metabolism of DEHT by the rat appears to occur via rapid hydrolysis of both ester linkages to give two moles of 2-ethylhexanol and one mole of terephthalic acid. Although 2-ethylhexanol has been shown to induce peroxisome proliferation, it appears to be less active in this respect than the monoester of DEHP. The relatively smaller amounts of monoester produced during the metabolism of DEHT may explain the differences seen in these experiments.

The involvement of the nervous system in systemic intoxication offers diagnostic and investigative challenges to the occupational health team. Often early evidence of neurotoxicity is vague, of a highly subjective nature and may be complicated by the presence of neuropathies due to other causes such as diabetes mellitus or exposure to multiple neurotoxins such as lead and solvents. The variety of agents producing neurotoxicity is wide, including heavy metals, organophosphates, acrylamide and its analogs, industrial solvents, therapeutic agents and plant alkaloids. The identification of neurotoxic substances requires well controlled epidemiologic studies of humans, including electrodiagnostic testing and well planned experimental animal studies. The latter include consideration of choice of animal species, routes of administration, duration of exposure, clinical evaluation, electrodiagnostic testing, morphologic description of the damage produced and its distribution, and the recognition of the potential for intercurrent naturally occurring neuropathologic processes. The aim of this discussion is to give an overview of these and other factors involved in the assessment of a neurotoxic potential.

Ethyl n-butyl ketone (EBK, 3-heptanone) has not been reported to produce neurotoxicity in previous studies when it was given by oral or inhalation routes. In the present study, EBK given by gavage at 2 g/kg/day, 5 days/week for 14 weeks produced a typical central-peripheral distal axonopathy characterized by "giant" axonal swelling and neurofilamentous hyperplasia. This dose approximates the single dose LD50 (2.76 g/kg) of EBK determined by Smyth et al., (J. Ind. Hyg. Toxicol. 31, 60-62, 1949). Large multiple doses (1.5 g/kg/day, 5 days/week, 14 weeks) of methyl ethyl ketone (MEK) given by gavage potentiated EBK neurotoxicity but 5-methyl-2-octanone did not. MEK modestly increased the urinary excretion of two neurotoxic gamma-diketones, 2,5-heptanedione, and 2,5-hexanedione, when MEK was given by gavage with EBK. When rats were exposed to EBK (700 ppm) and MEK (700 or 1400 ppm) in combination by inhalation exposure, serum 2,5-heptanedione levels were increased approximately 2.5 times. This effect was absent at MEK levels of 70 ppm. The serum from rats exposed to EBK or EBK/MEK combinations did not contain 2,5-hexanedione. The toxicity of EBK appears to involve the metabolism of EBK to two neurotoxic gamma-diketones, 2,5-heptanedione and 2,5-hexanedione. Combined EBK/MEK exposure modestly increased gamma-diketone levels in the serum and urine suggesting that MEK potentiates EBK neurotoxicity by stimulating the metabolism of EBK to neurotoxic metabolites. The magnitude of the doses used in the present study to produce neurotoxicity and the absence of neurotoxicity in previous subchronic studies suggest that the neurotoxic hazard of EBK is low.

3 ketone solvents (methyl ethyl ketone (MEK), methyl isobutyl ketone (MiBK), and isophorone) were tested for potential genotoxicity. The assays of MEK and MiBK included the Salmonella/microsome (Ames) assay, L5178Y/TK+/- mouse lymphoma (ML) assay, BALB/3T3 cell transformation (CT) assay, unscheduled DNA synthesis (UDS) assay, and micronucleus (MN) assay. Only the ML, UDS, and MN assays were conducted on samples of isophorone. No genotoxicity was found for MEK or isophorone. The presence of a marginal response only at the highest, cytotoxic concentration tested in the ML assay, the lack of reproducibility in the CT assay, and clearly negative results in the Ames assay, UDS and MN assays, suggest that MiBK is unlikely to be genotoxic in mammalian systems.

Tertiary butylhydroquinone (TBHQ), a phenolic antioxidant used in foods, was tested for its potential to produce lung damage in mice similar to that seen following administration of butylated hydroxytoluene (BHT). Male mice (CRL:CD-1) were given single intraperitoneal injections of 62.5, 125, 250 or 500 mg/kg TBHQ or 300, 625 or 1230 mg/kg BHT in corn oil. Survivors were killed five days after treatment and the lungs were removed, weighed, and processed for histologic examination. Lung weights of the TBHQ treated animals were comparable to the controls, while BHT produced a statistically significant increase in lung weight. Histologically, BHT produced hyperplasia of pulmonary pneumocytes as previously reported. No treatment-related lung lesions were seen in the TBHQ mice. The results of this study indicate that TBHQ does not produce pulmonary lesions in mice.

4-Chloro-3-nitroaniline was given to groups of 20 male and 20 female rats by gavage at doses of 3.6, 18, or 90 mg/kg in a 10% corn oil suspension. Doses were administered 5 days per week for 90 days. The high dose resulted in reduced body weight gain in males, reduced feed consumption and fluctuating feed consumption in both sexes, slight hemolytic anemia with Heinz bodies in both sexes, enlargement of the spleen due to congestion, hemosiderosis, and increased extramedullary hematopoiesis in both sexes, inflammatory changes in the splenic capsules of two females, hemosiderin pigmentation of the liver in both sexes, bone marrow hyperplasia in both sexes, increased liver weight in females, and testicular atrophy. The middle dose produced a fluctuating feed consumption pattern in males, Heinz bodies in both sexes, very slight anemia in females, splenic hemosiderosis in males, slightly increased splenic extramedullary hematopoiesis in females, hemosiderin pigmentation of the liver in males, and possibly increased liver weight in females, and inflammatory changes in the splenic capsule of one male. The low dose produced Heinz bodies in males and possibly females although the Heinz body counts of females were not statistically significant. The primary toxicologic damage produced by 4-chloro-3-nitroaniline is hemolytic or Heinz body anemia and testicular atrophy. Thus the toxicity of 4-chloro-3-nitroaniline is similar to that of other aromatic nitro and amino chemicals.