Abstract  An altered pattern of epigenetic modifications is central to many common human diseases, including cancer. Many studies have explored the mosaic patterns of DNA methylation and histone modification in cancer cells on a gene-by-gene basis; among their results has been the seminal finding of transcriptional silencing of tumour-suppressor genes by CpG-island-promoter hypermethylation. However, recent technological advances are now allowing cancer epigenetics to be studied genome-wide - an approach that has already begun to provide both biological insight and new avenues for translational research. It is time to 'upgrade' cancer epigenetics research and put together an ambitious plan to tackle the many unanswered questions in this field using epigenomics approaches.

Abstract  DNA methylation is a defining feature of mammalian cellular identity and is essential for normal development. Most cell types, except germ cells and pre-implantation embryos, display relatively stable DNA methylation patterns, with 70-80% of all CpGs being methylated. Despite recent advances, we still have a limited understanding of when, where and how many CpGs participate in genomic regulation. Here we report the in-depth analysis of 42 whole-genome bisulphite sequencing data sets across 30 diverse human cell and tissue types. We observe dynamic regulation for only 21.8% of autosomal CpGs within a normal developmental context, most of which are distal to transcription start sites. These dynamic CpGs co-localize with gene regulatory elements, particularly enhancers and transcription-factor-binding sites, which allow identification of key lineage-specific regulators. In addition, differentially methylated regions (DMRs) often contain single nucleotide polymorphisms associated with cell-type-related diseases as determined by genome-wide association studies. The results also highlight the general inefficiency of whole-genome bisulphite sequencing, as 70-80% of the sequencing reads across these data sets provided little or no relevant information about CpG methylation. To demonstrate further the utility of our DMR set, we use it to classify unknown samples and identify representative signature regions that recapitulate major DNA methylation dynamics. In summary, although in theory every CpG can change its methylation state, our results suggest that only a fraction does so as part of coordinated regulatory programs. Therefore, our selected DMRs can serve as a starting point to guide new, more effective reduced representation approaches to capture the most informative fraction of CpGs, as well as further pinpoint putative regulatory elements.

Abstract  Guinea-pig assays have been used extensively to detect contact sensitizers. In contrast, almost no reliable assays are available to detect the potential for low-molecular-weight drugs and chemicals to induce systemic allergic reactions in humans. Based on clinical data, and, to some extent, on recent immunological findings, it is proposed that guinea-pig assays can predict the hazard for systemic allergic reactions in man. Seventy drugs and chemicals were compared from published results in guinea-pig assays and in the clinic. A close correlation was found with 43 substances and a relatively good one with 16 substances. Conflicting results were found with 11 substances only. However, substances known to induce systemic allergic reactions in man were all detected as weak sensitizers, at least in guinea-pigs. Guinea-pig contact sensitization assays may therefore prove useful until more suitable and specific assays are available to predict the risk for systemic allergic rea

Abstract  Non-muscle-invasive bladder cancer (NMIBC) is heterogeneous, but current diagnostic and treatment strategies rely primarily on clinical parameters, lacking individualization to tumor and host genetics and biology. The heterogeneity of NMIBCs is derived from mutations, mutation signatures, chromosomal loss, and disruption of molecular pathways, which ultimately affects tumor progression, recurrence, and responsiveness to intravesical and systemic chemotherapy. Although research is still underway, advances in sequencing technology, insight into differential bacillus Calmette-Guérin responses, and new investigational treatment targets will soon offer clinicians new, precision-based tools to risk stratify and determine treatment regimens for future patients with bladder cancer.

Abstract  Increased oxidative stress and changes in DNA methylation are frequently detected in bladder cancer patients. We previously demonstrated a relationship between increased oxidative stress and hypomethylation of the transposable long-interspersed nuclear element-1 (LINE-1). Promoter hypermethylation of a tumor suppressor gene, runt-related transcription factor 3 (RUNX3), may also be associated with bladder cancer genesis. In this study, we investigated changes of DNA methylation in LINE-1 and RUNX3 promoter in a bladder cancer cell (UM-UC-3) under oxidative stress conditions, stimulated by challenge with H2O2 for 72 h. Cells were pretreated with an antioxidant, tocopheryl acetate for 1 h to attenuate oxidative stress. Methylation levels of LINE-1 and RUNX3 promoter were measured by combined bisulfite restriction analysis PCR and methylation-specific PCR, respectively. Levels of LINE-1 methylation were significantly decreased in H2O2-treated cells, and reestablished after pretreated with tocopheryl acetate. Methylation of RUNX3 promoter was significantly increased in cells exposed to H2O2. In tocopheryl acetate pretreated cells, it was markedly decreased. In conclusion, hypomethylation of LINE-1 and hypermethylation of RUNX3 promoter in bladder cancer cell line was experimentally induced by reactive oxygen species (ROS). The present findings support the hypothesis that oxidative stress promotes urothelial cell carcinogenesis through modulation of DNA methylation. Our data also imply that mechanistic pathways of ROS-induced alteration of DNA methylation in a repetitive DNA element and a gene promoter might differ.

Abstract  The rapidly expanding field of medical geology deals with the relationships between natural geological factors and health, both human and animal. It also aims to improve our understanding of the ways in which the geological environment has an impact on the geographical distribution of health problems. This new book brings together the work of geoscientists and medical/public health researchers, and addresses the health problems caused, or exacerbated, by geological materials (rocks, minerals, atmospheric dust and water) and processes (including volcanic eruptions and earthquakes). Among the environmental health problems discussed in the volume are: human and animal exposure to toxic levels of trace essential and non-essential elements such as arsenic and mercury; trace element deficiencies; exposure to natural dusts and to radioactivity; naturally occurring organic compounds in drinking water; and the effects of volcanic emissions. Examining the positive side of the equation as well as the negative, the book also deals with the many health benefits of geologic materials and processes. This wide-ranging volume covers issues in medical geology all over the world with each author covering their respective region. It provides examples from different continents as well as a state-of-the-art review of the latest developments in the discipline. The authors are all recognized geoscientific and medical experts working in the field. The book is written for a wide variety of specialists from geologists, geochemists, pathologists and medical doctors to veterinarians and biologists.

Abstract  Arsenic is a known potent risk factor for bladder cancer. Increasing evidence suggests that epigenetic alterations, e.g., DNA methylation and histones posttranslational modifications (PTMs), contribute to arsenic carcinogenesis. Our previous studies have demonstrated that exposure of human urothelial cells (UROtsa cells) to monomethylarsonous acid (MMAIII), one of arsenic active metabolites, changes the histone acetylation marks across the genome that are correlated with MMAIII-induced UROtsa cell malignant transformation. In the current study, we employed a high-resolution and high-throughput liquid chromatography tandem mass spectrometry (LC-MS/MS) to identify and quantitatively measure various PTM patterns during the MMAIII-induced malignant transformation. Our data showed that MMAIII exposure caused a time-dependent increase in histone H3 acetylation on lysine K4, K9, K14, K18, K23, and K27, but a decrease in acetylation on lysine K5, K8, K12, and K16 of histone H4. Consistent with this observation, H3K18ac was increased while H4K8ac was decreased in the leukocytes collected from people exposed to high concentrations of arsenic compared to those exposed to low concentrations. MMAIII was also able to alter histone methylation patterns: MMAIII transformed cells experienced a loss of H3K4me1, and an increase in H3K9me1 and H3K27me1. Collectively, our data shows that arsenic exposure causes dynamic changes in histone acetylation and methylation patterns during arsenic-induced cancer development. Exploring the genomic location of the altered histone marks and the resulting aberrant expression of genes will be of importance in deciphering the mechanism of arsenic-induced carcinogenesis.

Abstract  Environmental arsenic exposure, through drinking contaminated water, is a significant risk factor for developing vascular diseases and is associated with liver portal hypertension, vascular shunting, and portal fibrosis through unknown mechanisms. We found that the addition of low doses of arsenite to the drinking water of mice resulted in marked pathologic remodeling in liver sinusoidal endothelial cells (SECs), including SEC defenestration, capillarization, increased junctional PECAM-1 expression, protein nitration, and decreased liver clearance of modified albumin. Furthermore, the pathologic changes observed after in vivo exposure were recapitulated in isolated mouse SECs exposed to arsenic in culture. To investigate the role of NADPH oxidase-generated ROS in this remodeling, we examined the effect of arsenite in the drinking water of mice deficient for the p47 subunit of the NADPH oxidase and found that knockout mice were protected from arsenite-induced capillarization and protein nitration. Furthermore, ex vivo arsenic exposure increased SEC superoxide generation, and this effect was inhibited by addition of a Nox2 inhibitor and quenched by the cellpermeant superoxide scavenger. In addition, inhibiting either oxidant generation or Rac1-GTPase blocked ex vivo arsenic-stimulated SEC differentiation and dysfunction. Our data indicate that a Nox2-based oxidase is required for SEC capillarization and that it may play a central role in vessel remodeling following environmentally relevant arsenic exposures. [ABSTRACT FROM AUTHOR] Copyright of Journal of Clinical Investigation is the property of American Society for Clinical Investigation and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts)

Abstract  This laboratory has shown that both Cd(+2) and As(+3) can malignantly transform human urothelial cells. The present study examined metal resistance and the mechanism of cell death when the parental and malignantly transformed UROtsa cells were exposed to Cd(+2) and As(+3). It was shown that the malignantly transformed UROtsa cells were more resistant to the toxic effects of both metals. The assessment of the mode of cell death demonstrated that the parental UROtsa cells died by both apoptosis and necrosis when exposed to either metal. It was shown that apoptosis was the more prominent mechanism of cell death, accounting for over 50% of cell death. Apoptotic cell death was determined by the observation of fragmented nuclei using 4',6-diamidino-2-phenylindole staining, the formation of a DNA ladder, and the detection of cleaved caspase-3 and caspase-9 products in the cell lysates. Necrotic cell death was determined by measuring the release of lactate dehydrogenase into the growth medium. It was determined that the extent of apoptosis of the malignantly transformed UROtsa cells was decreased and that the extent of necrosis was increased compared to the parental UROtsa cells. These observations are consistent with in vivo studies which suggest that As(+3) can act as a tumor promoter during the regeneration of the bladder urothelium. The present in vitro studies suggest that As(+3)-induced cytotoxicity could set the stage for tissue repair due to its own inherent toxicity to normal urothelium, and then subsequently act as a tumor promoter during the regeneration process through the stimulation of the regrowth of cells that have gained increased resistance to As(+3).

Abstract  Carcinogenesis is an important chronic toxicity of metals and metalloids, although their mechanisms of action are still unclear. Comparison of gene expression patterns induced by carcinogenic metals, metalloids, and model carcinogens would give an insight into understanding of their carcinogenic mechanisms. In this study, we examined the gene expression alteration in human hepatoma cell line, HepG2, after exposing to two metals (cadmium and nickel), a metalloid (arsenic), and three model carcinogenic chemicals N-dimethylnitrosoamine (DMN), 12-O-tetradecanoylphorbol-13-acetate (TPA), and tetrachloroethylene (TCE) using DNA microarrays with 8,795 human genes. Of the genes altered by As, Cd, and Ni exposures, 31-55% were overlapped with those altered by three model carcinogenic chemical exposures in our experiments. In particular, the metals and metalloid shared certain characteristics with TPA and TCE in remarkable upregulations of the genes associated with progression of cell cycle, which might play a central role in As, Cd, and Ni carcinogenesis. This characteristic of gene expression alteration was partially counteracted by intracellular accumulation of vitamin C in As-exposed cells, whereas the number of cell-cycle associated genes was increased in Cd- and Ni-exposed cells. In our experimental conditions, ROS might have an accelerative effect on the cell proliferation mechanisms of As, but have an inhibitory effect on those of other two heavy metals. Furthermore, based on the results of Q-PCR, the oncogene PTTG1, which was upregulated by all carcinogenic chemical exposures in the array experiments, might be a useful biomarker for evaluation of the carcinogenesis of inorganic carcinogens

Abstract  Biological monitoring of occupational exposure is characterized by important variability, due both to variability in the environment and to biological differences between workers. A quantitative description and understanding of this variability is important for a dependable application of biological monitoring. This work describes this variability, using a toxicokinetic model, for a large range of chemicals for which reference biological reference values exist. A toxicokinetic compartmental model describing both the parent compound and its metabolites was used. For each chemical, compartments were given physiological meaning. Models were elaborated based on physiological, physicochemical, and biochemical data when available, and on half-lives and central compartment concentrations when not available. Fourteen chemicals were studied ( arsenic, cadmium, carbon monoxide, chromium, cobalt, ethylbenzene, ethyleneglycol monomethylether, fluorides, lead, mercury, methyl isobutyl ketone, penthachlorophenol, phenol, and toluene), representing 20 biological indicators. Occupational exposures were simulated using Monte Carlo techniques with realistic distributions of both individual physiological parameters and exposure conditions. Resulting biological indicator levels were then analyzed to identify the contribution of environmental and biological variability to total variability. Comparison of predicted biological indicator levels with biological exposure limits showed a high correlation with the model for 19 out of 20 indicators. Variability associated with changes in exposure levels ( GSD of1.5 and 2.0) is shown to be mainly influenced by the kinetics of the biological indicator. Thus, with regard to variability, we can conclude that, for the 14 chemicals modeled, biological monitoring would be preferable to air monitoring. For short half-lives ( less than 7 hr), this is very similar to the environmental variability. However, for longer half-lives, estimated variability decreased.

Abstract  BACKGROUND: Altered patterns of gene expression mediate the effects of particulate matter (PM) on human health, but mechanisms through which PM modifies gene expression are largely undetermined. MicroRNAs are highly conserved, non-coding small RNAs that regulate the expression of broad gene networks at the post-transcriptional level. OBJECTIVES: We evaluated the effects of exposure to PM and PM metal components on candidate miRNAs (miR-222, miR-21 and miR-146a) related with oxidative stress and inflammatory processes in 63 workers of an electric-furnace steel plant. METHODS: We measured miR-222, miR-21 and miR-146a expression in blood leukocyte RNA on the first day of a workweek (baseline) and after three days of work (post-exposure). Relative expression of microRNAs was measured by real-time PCR. We measured blood oxidative stress (8-hydroxyguanine) and estimated individual exposures to PM1, PM10, coarse particles and PM metal components (chromium, lead, cadmium, arsenic, nickel, manganese) between the baseline and post-exposure measurements. RESULTS: miR-222 and miR-21 expression was significantly increased in post-exposure samples (baseline=0.68+/-3.41, post-exposure=2.16+/-2.25; p=0.002 for miR-222; baseline=4.10+/-3.04, post-exposure=4.66+/-2.63,; p=0.05 for miR-21). In post-exposure samples, miR-222 expression was positively correlated with lead exposure (beta=0.41, p=0.02), while miR-21 expression was associated with blood 8-hydroxyguanine (beta=0.11, p=0.03), but not with individual PM or metals. Post-exposure expression of miR-146a was not significantly different from baseline (baseline=0.61+/-2.42, post-exposure=1.90+/-3.94; p=0.19), but it was negatively correlated with exposure to lead (beta=-0.51, p=0.011) and cadmium (beta=-0.42, p=0.04). CONCLUSIONS: Changes in miRNA expression may represent a novel mechanism mediating responses to PM and its metal components.

Abstract  VA Merit Review grants; National Institutes of Health frants HL-36045 and HL-02311; American Heart Association, Texas Affiliate. #Background Carbon monoxide (CO), like nitric oxide (NO), stimulates soluble guanylyl cyclase and thereby raises intracellular levels of cGMP. We examined the endogenous capacity of vascular smooth muscle cells (SMCs) to produce CO from heme through the activity of heme oxygenases. Methods and Results Cultured SMCs from rat aorta (RASMCs) expressed immunoreactive inducible heme oxygenase-1 (HO-1) and constitutive HO-2. Treatment of RASMCs with hemin and sodium arsenite, which are inducers of HO-1, stimulated RASMC cGMP without stimulating nitrite release or inducible NO synthase expression, and the induced elevations of cGMP were not inhibited by the NO synthase inhibitor NG-methyl-L-arginine. Induced CO from RASMCs likewise caused elevation of cGMP levels in platelets coincubated with the vascular cells. Zinc protoporphyrin IX, an inhibitor of HO, reversed the inducible increases in platelet cGMP. Conclusions These results indicate that vascular SMCs have both constitutive and inducible HO activity, and they respond to specific stimuli to generate guanylyl cyclaseûstimulatory CO in the same SMCs and in coincubated platelets.

Abstract  Female Swiss mice were exposed to sodium arsenite or sodium aresenate in the drinking water for 15 weeks at concentrations ranging from 0 to 100 micrograms/mL arsenic content. After three weeks of the 15 week exposure period, the mice were administered urethan (1.5 mg/g) intraperitoneally. Pulmonary adenoma formation was evaluated 12 weeks later. Arsenic exposure produced a protective effect with respect to tumor development. Both forms of arsenic reduced the size and number of pulmonary adenomas observed per mouse. In addition, urethan-induced sleeping times which reflect the rate of urethan metabolism or excretion remained unchanged. This suggests that arsenic exposure does not alter urethan excretion and is not a factor influencing subsequent adenoma formation of these levels of exposure.

Abstract  The transfer of 73As and 125SB to the developing mouse organism was studied in the following ways. a)A diet containing the isotopes was given from the day of the vaginal plug and the radioactivity in uterus, ovaries and different maternal organs was determined 2,4, and 6 days after initiation of feeding. b)The isotopes were injected i.p. during organogenesis at day 12, and the transfer into the fetus as well as activity in maternal organs were followed for a period of 7 days. c)The radioactive diet was supplied during the entire pregnancy and 15 days thereafter. The radioactivity in the females was followed for 150 days, that in the litter for 50 days. Components of turnover of less than a day, less than a week and several months can be discerned in the adult for both isotopes and transfer to the fetus occurs during organogenesis. Less antimony than arsenic is transferred from food during fetal life. After birth, arsenic as well as antimony are taken up by the newborn via the milk.

Abstract  Splenic macrophages are highly efficient in trapping and concentrating foreign substances carried in the blood and also the major sites where antibodies are synthesised and from where they are released into the circulation. Lead and Arsenic as environmental agents are considered to be high priority toxic substances largely due to their carcinogenic potentials in humans. However, these heavy metals as carcinogens remain an enigma because while they are definitely active in humans, carcinogenesis in the rodent model has never been convincingly demonstrated. Although macrophages are predominantly recruited to the site of inflammation during inflammatory distress as well as in immune response, nothing is known about the interaction of lead and arsenic with macrophages and their possible role in immunotoxicologic effect. In the present study it is reported that in vivo lead acetate treatment (10 mg/kg body wt) inhibits the cell adhesion property and alters the cell morphology in the splenic macrophages. Results show that there is a significant decrease in alkaline phosphatase release in lead treated macrophages (6.7 +/- 0.88 IU/100 mL) with respect to control (14.17 +/- 0.18). In vivo exposure of sodium arsenite (0.5 mg/kg body wt) also decreases phagocytic activity for ingestion and digestion of exogenous antigens, such as whole microorganism, as evident from the phagocytic index, 11555.55 +/- 62.86 (in control) to 5555.5 +/- 1571.33 in arsenic treated cells. Arsenic exposed cells release 8.15 +/- 0.05 microM nitric oxide, whereas control cells release 10.95 +/- 0.15 microM of nitric oxide, which is also identical following LPS stimulation. Results show that the functional integrity of the target cell is also decreased after arsenic exposure as obtained from the percentage of DNA fragmentation. A greater percentage of DNA fragmentation upon arsenic treatment (43.1 +/- 0.05%) with respect to control (14.9 +/- 0.34%) indicates that arsenic induces apoptosis. In immune cells which are rapidly proliferating and differentiating, inhibition of these heavy metal induced functions may result in similar degree of toxicity and lead to diseased state.

Abstract  Several studies have found elevated levels of urinary arsenic among residents living near a copper smelter in Tacoma, Washington. To assess pathways of exposure to arsenic from the smelter, biological and environmental samples were collected longitudinally from 121 households up to 8 miles from the smelter. The concentration of inorganic and methylated arsenic compounds in spot urine samples was used as the primary measure of exposure to environmental arsenic. Urinary concentration of arsenic dropped off to a constant background level within one-half mile of the smelter in contrast to environmental concentrations, which decreased more steadily with increasing distance. Among all age-sex-specific groups in all areas, only children ages 0-6 living within one-half mile of the smelter had elevated levels of arsenic in urine. A separate analysis of data for these children suggests that hand-to-mouth activity was the primary source of exposure. Inhalation of ambient air and resuspension of contaminated soil were not important sources of exposure for children or adults.

Abstract  Mining and smelting at Kellogg-Smelterville, Idaho, resulted in high concentrations of lead in Coeur d'Alene (CDA) River sediments 15-65 km downstream, where ospreys (Pandion haliaetus) nested. Adult and nestling ospreys living along the CDA River had significantly higher lead concentrations than those at Lake Coeur d'Alene (intermediate area) or Pend Oreille and Flathead Lakes (reference areas). Lead concentrations in fish collected from the sandy areas paralleled those found in ospreys. Inhibition of blood .delta.-aminolevulinic acid dehydratase (ALAD) activity and elevation of protoporphyrin concentration provided evidence of lead exposure. In adult ospreys, ALAD activity was negatively correlated with lead in blood (r = -0.57), whereas protoporphyrin was positively correlated with lead in blood (r = +0.40). Neither hemoglobin nor hematocrit was adversely affected by the relatively modest lead concentrations found in the blood. Pronounced accumulation of lead by adults or young could ultimately result in behavioral abnormalities or death, both of which would reduce productivity of the nesting osprey population. We did not observe death related to lead, behavioral abnormalities, or reduced productivity during this 1986-87 study. Despite some lead-induced biochemical changes in blood parameters, ospreys produced young at nearly identical rates in the three study areas; these rates were among the highest ever reported in the western United States. Post-fledging survival of ospreys exposed to lead early in life remains an unknown. Lead does not biomagnify in the food chain as do organochlorine pesticides and mercury and several osprey behavior traits reduce the potential for the species to accumulate critical levels of lead. Swans, which feed at a lower trophic level, continue to die from environmental lead in the region.

Abstract  The occurrence of chromosome aberrations was studied in short-term cultured lymphocytes from nine workers exposed to arsenic at the Rönnskär smeltery in northern Sweden. In the smelter workers, 87 aberrations were found in 819 mitoses. The number of aberrations varied individually from 0 to 25 aberrations per 100 cells. In a control material 13 aberrations were found in 1012 mitoses. The frequency of chromosome aberrations was significantly increased among the smelter workers, but due to the simultaneous exposure to other agents the effect of arsenic per se can not be assessed with certainty.

Abstract  An investigation on the relationship between dietary arsenic exposure and cardiovascular diseases was made. In Antofagasta Commune, northern Chile, since 1955 arsenic has polluted public drinking water. This environmental contamination is of geological origin. The concentration of arsenic in drinking water for the 1955-1970 period was 0.5980 ppm (weighted mean). In the period June 1970-March 1972, the concentration decreased to 0.0815 ppm (weighted mean), due to a Water Filtration Plant which started operating in May 1970. Greater Santiago showed 0.00 ppm of arsenic in drinking water. Amongst 10 autopsied patients (age range: 1 year 7 months to 18 years) with chronic arsenical dermatosis from Antofagasta Commune, 9 showed marked fibrous intimal thickening of the arterial wall and/or restricted lumen of the left coronary artery, 2 of these 9 also exhibiting myocardial infarction. Of the 10 patients, 7 developed cardiomegaly, which was related to chronic exposure to dietary arsenic. Two series of patients with myocardial infarction under 40 years of age, one from Antofagasta Commune (exposed to arsenic), the other from Greater Santiago (not exposed to arsenic) were compared. The Yates' corrected chi 2 value (1 d.f.) being 11.7776. The difference was statistically highly significant (P approximately equal to 0.0006018). Furthermore, in Antofagasta Commune, the number of cases (< 40 yr) which had myocardial infarction with chronic arsenical dermatosis were compared with the cases (< 40 yr) which showed myocardial infarction without chronic arsenical dermatosis. The Yates' corrected chi 2 value (1 d.f.) was 13.0395. A highly significant difference was detected (P approximately equal to 0.0003044). Children (0-15 yr)from the two cities were also compared.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract  Monomethylarsonous acid (MMAIII), a trivalent metabolite of arsenic, is highly cytotoxic and recent cell culture studies suggest that it might act as a carcinogen. The general consensus of studies indicates that the cytotoxicity of MMAIII is a result of increased levels of reactive oxygen species (ROS). A longstanding relationship between arsenic and selenium metabolism has led to the use of selenium as a supplement in arsenic exposed populations, however the impact of organic arsenicals (methylated metabolites) on selenium metabolism is still poorly understood. In this study we determined the impact of exposure to MMAIII on the regulation of expression of TrxR1 and its activity using a primary lung fibroblast line, WI-38. The promoter region of the gene encoding the selenoprotein thioredoxin reductase 1 (TrxR1) contains an antioxidant responsive element (ARE) that has been shown to be activated in the presence of electrophilic compounds. Results from radiolabeled selenoproteins indicate that exposure to low concentrations of MMAIII resulted in increased synthesis of TrxR1 in WI-38 cells, and lower incorporation of selenium into other selenoproteins. MMAIII treatment led to increased mRNA encoding TrxR1 in WI-38 cells, while lower levels of mRNA coding for cellular glutathione peroxidase (cGpx) were detected in exposed cells. Luciferase activity of TrxR1 promoter fusions increased with addition of MMAIII, as did expression of a rat quinone reductase (QR) promoter fusion construct. However, MMAIII induction of the TRX1 promoter fusion was abrogated when the ARE was mutated, suggesting that this regulation is mediated via the ARE. These results indicate that MMAIII alters the expression of selenoproteins based on a selective induction of TrxR1, and this response to exposure to organic arsenicals that requires the ARE element.

Abstract  Inorganic arsenic enhances skin tumor formation when combined with other carcinogens including ultraviolet radiation (UVR). The inhibition of DNA damage repair by arsenic has been hypothesized to contribute to the cocarcinogenic activities of arsenic observed in vivo. Cyclobutane pyrimidine dimers (CPDs) are an important mutagenic UVR photoproduct and implicated in the genesis of nonmelanoma skin cancer. The current study demonstrates that low concentrations of arsenite (As(III)) inhibit UVR-induced CPD repair in a human keratinocyte cell line via nitric oxide (NO) and inducible nitric oxide synthase (iNOS). Following As(III) treatment, NO production and iNOS expression are elevated. Little is known about regulation of iNOS by As(III) and further investigations indicated that p38 mitogen-activated protein kinase (p38 MAPK) and NF-kappaB are required for As(III) induction of iNOS expression. This As(III)-stimulated signaling cascade was involved in inhibition of UVR-induced CPD repair as disruption of p38 MAPK activity and NF-kappaB nuclear translocation counteracted the effects of As(III) on CPD repair. Selective inhibition of iNOS ameliorated As(III) inhibition of CPD repair, thereby suggesting that iNOS is a downstream mediator of As(III) activity. These findings provide evidence that an As(III)-stimulated signal transduction cascade culminating in elevated iNOS expression and NO generation is an underlying mechanism for inhibition of UVR-induced DNA damage repair by arsenic.

Abstract  ABCC5/MRP5 is an organic anion transporter that participates in tissue defense and cellular signal transduction through efflux of anticancer drugs, toxicants and a second messenger cGMP, but its physiological functions in zebrafish remain to be defined. Herein, we report the characterization, spatiotemporal expression and developmental function of zebrafish ABCC5 and its transcriptional responses to heavy metals. Zebrafish abcc5 gene is located on chromosome 18 and comprised of 28 exons. The deduced polypeptide of zebrafish ABCC5 consists of 1426 amino acids, which shares high sequence identity with those from other species. Zebrafish abcc5 is maternally expressed and its transcripts are mainly distributed in brain, lens, liver and intestine of developing embryos. In adults, zebrafish abcc5 is extensively expressed, at higher levels in testis, brain, eye, ovary, intestine and kidney, but at relatively lower levels in gill, liver, heart and muscle. Blockage of endogenous ABCC5 activity by its dominant-negative led to the developmental retardation of zebrafish embryos in which activity of p21 signaling was markedly stimulated and cellular cGMP content was significantly increased. In addition, expression of abcc5 in ZF4 cells and zebrafish embryos was significantly induced by cadmium (Cd), lead (Pb), mercury (Hg) or arsenic (As). The induced expression of ABCC5 by heavy metals was mainly detected in the liver of embryos at 96-h post-fertilization (hpf). In adult zebrafish, expression of abcc5 in brain, intestine, liver, kidney and ovary was significantly induced by one or more of these heavy metals. Furthermore, overexpression of ABCC5 attenuated the toxicity of Cd to zebrafish embryos, but did not affect the toxicity of Hg or As. Thus, ABCC5 is likely to play an active role in embryonic development and heavy metal detoxification through the export of second messenger molecules and toxicants out of cells in zebrafish.

Abstract  Data on the mechanism of antimony genotoxicity is scarce. Arsenic and antimony are proposed to share some toxicological features. Thus comparative and combined experiments with As(III) and Sb(III) were performed to gain a deeper knowledge of the mechanism of antimony genotoxicity. Trivalent arsenic proved to be five times more cytotoxic and one order of magnitude more potent in induction of micronuclei in human lymphocytes in vitro than was antimony. Significantly increased micronucleus frequencies were achieved with As(III) at a dose of 0.5 microM and with Sb(III) at a dose of 5 microM. Neither the number of micronuclei induced by As(III) nor by Sb(III) could be suppressed by co-incubation with superoxide dismutase or catalase. This suggests that induction of oxidative stress may not be a crucial step in the mechanism of DNA damage induction by arsenic and antimony. The combined genotoxicity in micronucleus test co-incubation experiments with arsenic and antimony seemed best described by simple additivity. In the single cell gel test with human lymphocytes a significant induction of DNA damage was observed with 0.01 microM As(III) and 5 microM Sb(III). In contrast to Sb(III), As(III) proved to be a very potent inducer of DNA-protein crosslinks. It may be that Sb(III) as well as As(III) causes DNA damage by inhibition of enzymes involved in DNA repair. Further investigations will have to identify the relevant sites of action.