Abstract  BIOSIS COPYRIGHT: BIOL ABS. RRM ABSTRACT HUMAN SUICIDE

Abstract  Aplastic anemia developed in a 67-year-old man after many months' use of arsenic-containing pesticides. He was treated with penicillamine and oxymetholone, which promptly restored his blood count to normal limits. Three months later he again had aplastic anemia, and the urine once more contained considerable amounts of arsenic. Treatment with pencillamine and oxymetholone was reinstituted. This resulted in excellent urinary excretion of arsenic, but his blood count did not improve. Five months after pancytopenia first developed, he had acute myelogenous leukemia, which was treated with prednisone, cytosine arabinoside, and thioguanine. This had no beneficial effects, and he died of klebsiella septicemia. Autopsy showed widespread leukemic infiltrates.

Abstract  The biotransformation, tissue retention, intracellular binding and biokinetics of arsenic were studied in rabbits exposed to [74As]arsenate (0.4 mg As/kg body wt., i.v.). Inhibition of the methyltransferase activity by injection of periodate-oxidized adenosine (PAD) caused a marked decrease of the formation of [74As]dimethylarsinic acid (DMA), which gave rise to 1.5–4 times increased tissue levels of 74As. This is almost the same as reported for rabbits given arsenite in combination with PAD and was due to a rapid reduction of the arsenate to arsenite which bound to the tissues. Only about 30% of the arsenate given was excreted unchanged in the urine, indicating that a large part was reduced to AsIII. Thus the methylation to DMA seems to be almost as important for the detoxication following exposure to arsenate as that following exposure to arsenite. In the rabbits with normal methylating capacity 50–70% of the produced AsIII was methylated to DMA. The liver was the only organ in which DMA was present 1 h after the administration, indicating that this is the main site of the methylation. The DMA was rapidly cleared from all tissues except the thyroid.

Abstract  The accumulation of arsenic with time in tissues of rats and rabbits was determined by neutron activation analysis (NAA). Rats showed a steady increase in the As-concentrations with age, whereas in rabbits it was nearly the same for adults and in young animals. The metabolism of arsenic was studied in both animal species after i.p. injection of 50/micrograms As/kg b.w. as 74As labelled arsenite. Eight tissues, as well as blood and urine, were analysed for 74As content after 16 and 48 hours. The binding of 74As to hematic and intracellular components and the chemical forms of arsenic in tissues and urine were investigated. In the plasma and the RBC-fraction of the rabbit, the As concentration decreased during the first two days, while in the rats it only disappeared from the plasma, but was retained in the RBC-fraction. Liver, kidney and lung of rabbits with the highest As concentrations at 16 and 48 hours showed a rapid clearance of As in the first 48 hours. In the corresponding tissues of the rats, the rate of decline was significantly lower, due to the higher binding of 74As to tissue constituents. Poor binding of As to plasma proteins was seen in rabbits while in rats it was totally bound to this fraction. In the RBC, liver and kidney cytosols, however, the affinity of As for intracellular proteins was higher in both animal species but characterized by a rate of binding different between the two animal species. The amount of dimethylarsinic acid (DMA) in the tissues was significantly lower in the rat than in the rabbit, reflecting the total amount of diffusible arsenic, which was also much lower in the tissues of rats than in rabbits.

Abstract  The carcinogenicity of inorganic arsenic in humans, particularly in the lung and skin, has been reasonably well established through epidemiological investigations. However, there is no substantial experimental evidence for carcinogenicity in animals to suort the human studies. Studies of metabolism and disposition of inorganic arsenic in various animal species are particularly relevant to determining the factors that might account for the lack of an animal model. Numerous studies of this type have been reported, but there do not appear to be clear qualitative or quantitative differences in the overall fate and disposition of inorganic arsenic in most animals versus humans, although little is known at the cellular and subcellular level. Sulphur chemistry, especially thiol status, is emerging as an important regulating factor in the overall fate and distribution of inorganic arsenic in the body, playing a role in the initial reduction of arsenate to arsenite and subsequent methylation, and possibly in determining tissue affinity and distribution properties. The metabolism of inorganic arsenic can be viewed as a redox cycle in which thiol compounds such as glutathione (GSH) possibly function as reducing agents and methyl donors as oxidising agents. One explanation for the possible sensitivity of certain malnourished human populations to the carcinogenic effects of inorganic arsenic may be related to the reduced availability of nonprotein sulphhydryl compounds such as GSH needed to drive the redox cycle and facilitate arsenic detoxification. Future carcinogenicity studies of inorganic arsenic in animals could be designed to address directly this aspect of the problem.

Abstract  The relative toxicity of gallium arsenide (GaAs) and its metal oxides was assessed by intratracheally instilling particulate suspensions of GaAs (100 mg/kg), equimolar gallium as Ga2O3 (65 mg/kg), or a maximally tolerated nonlethal dose of arsenic as As2O3 (17 mg/kg). Two weeks after dosing, five rats from each group were randomly selected for the biochemical determination of lung lipid, protein, DNA, and collagen (4-hydroxyproline; 4-HP) content. The pulmonary retention of gallium and/or arsenic and the concentration of these metals in blood were also determined. Lungs from the remaining rats (n = 3) were examined histopathologically. Pulmonary exposure to Ga2O3 particulates significantly (p less than 0.05) increased the total lipid content of lungs relative to that observed in the vehicle-treated control animals. This response appeared to be associated with the pulmonary retention of gallium particulates (means = 36% of the gallium dose). In contrast, As2O3 particulates were not retained in the lung. Blood arsenic concentrations were 36 ppm which represented 20% of the total arsenic administered. Treatment with As2O3 significantly elevated lung dry weight as well as protein, DNA, and 4-HP content. These data suggest that As2O3 induced an acute fibrogenic response. The intratracheal instillation of GaAs particulates produced effects similar to those observed with the individual oxides. The total lung content of lipids, protein, and DNA was significantly elevated. These biochemical changes were accompanied by significant increases in lung dry weight and lung wet weight. Lungs from rats receiving GaAs particulates retained 44% of the dose as gallium and 28% of the dose as arsenic at the end of the 14-day study. Blood arsenic concentrations were 44 ppm (7% of the arsenic dose) while gallium was not detected in blood at this time. The primary histopathological observations 14 days after the intratracheal instillation of all metal particulates were an inflammatory response and pneumonocyte hyperplasia. The biological severity of these lesions, in descending order, was GaAs greater than As2O3 much greater than Ga2O3. It must be noted, however, that As2O3 was dosed at 0.25 X moles of GaAs.

Abstract  A patient with arsenic intoxication is reported, who presented with a variety of gastrointestinal and neurologic disturbances including unilateral facial nerve palsy and acute symptomatic pancreatitis, neither of which have been previously described as sequelae of arsenic poisoning. The patient also suffered hematologic, dermatologic, and cardiopulmonary complications. A review of the literature about this interesting problem is also presented.

Abstract  To obtain the usual values of arsenic, beryllium, bismuth, cadmium, chromium, cobalt, copper, mercury, methyl mercury, manganese, molybdenum, nickel, lead, antimony, vanadium, and zinc in the normal human body, the amounts of 15 metals were determined in 15 male and 15 female Japanese cadavers (average weight, 55 kg [121 lb]). The content of metals found ranged as follows: Zn, 1,800 mg; Cu, 65 mg; Cd, 35 mg; Pb, 25 mg; Mn, 8 mg; Ni, 6 mg; Cr, 4 mg; Hg, 3 mg; Sb, 0.7 mg; MeHg, 0.4 mg. Cadmium and mercury were higher in Japanese blood than in blood of other people. Cadmium and mercury were absorbed by the metabolic tissues; Cr, Ni, and Pb showed higher concentration in tissue exposed to the environment. Concentrations of Cd, Pb, and Hg tended to be higher in females, and Cr Cu, MeHg, and Mn concentrations tended to be higher in males.

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 toxicity of Hg2+ [HgCl2], Cr6+(1) [(NH4)2CrO4], Cr6+(2) [CrO3], Cd2+ [CdCl2.2,5H2O], Pb2+ [Pb(CH3COO)2.3H2O], and As5+ [Na2HAsO4.7H2O] on the sensitivity and survival of Daphnia magna and Tubifex tubifex has been studied. All test metals were dissolved and determined under standardized conditions (dilution water, Bringmann and Kuhn, 1982) and 96 hr LC50 for T. tubifex and 48 hr LC50 for D. magna were compared in rank orders toxicity. For D. magna rank order toxicity was Hg2+ > Cr6+(2) > Cd2+ = Cr6+(1) > Pb2+ > As5+ and for T. tubifex it was Hg2+ > Cd2+ > Cr6+(2) > Cr6(1) > Pb2+ > As5+. D. magna was a more sensitive organism than T. tubifex and its LC50 values for all metals were several times lower than LC50 values for T. tubifex. Correlation between toxicity of various metals and biological subjects may be useful in predicting toxicity to various biologically important organisms connected with food chains.

Abstract  The effects of metal salts on Rauscher leukemia virus (RLV) were investigated in CD-1-mice. Mice were given intraperitoneal (ip) injections of 300 plaque forming units of virus at 6 weeks of age. Between the ages of 4 and 14 weeks, animals received drinking water containing salts of arsenic (7440382), cadmium (7440439), cobalt (7440484), mercury (7439976), nickel (7440020), lead (7439921), copper (7440508), and iron (7439896). At weeks 3 and 6 of treatment, mice were killed and spleens removed. Suspensions were prepared, and virus titrations were performed. Treatment of mice with cobalt-sulfate (10124433) and lead-acetate (301042) induced RLV splenomegalies. High titers of virus were found in larger spleens. Average spleen weight 6 weeks after lead treatment in RLV infected mice was 7 times greater than in those with virus alone. Spleens of RLV infected mice treated with mercuric-chloride (7487947) were larger than those of mice with virus alone. Spleen weights in RLV infected mice treated with cadmium, cobalt, iron, lead, and sodium-arsenite (7784465) were larger at 6 weeks than at 3 weeks and were larger than those in mice with virus alone. Spleens of lead treated RLV infected mice had high titers of virus; spleens treated with cobalt and mercury had high titers, but not as high as with lead. Spleens of mice treated with metals alone were not different from nonmetal treated, nonvirus injected controls. Nonmetal treated, virus injected mice did not have detectable virus in spleens. The author concludes that specific reasons for the interactions between metal and RLV treatment are not known.

Abstract  Epidemiological data relating arsenic ingestion and skin and internal cancers strongly suggest a sublinear or threshold relationship. Physiological saturation of methylation-based arsenic detoxification has been proposed as one explanation for a sublinear response. We have evaluated the molecular bases for sublinearity in light of new data and hypotheses regarding arsenic genotoxicity and chemical carcinogenesis. A review of the dose–response relationships observed in arsenic genotoxicity assays is presented. With the exception of sister chromatid exchanges, sublinear dose–response relationships for arsenic-induced chromosomal aberrations were observed repeatedly in different mammalian and human cell systems. Arsenic also enhanced the clastogenicity and mutagenicity of other DNA damaging agents with a sublinear dose response. Consistent with the dose response of arsenic-induced genetic alterations, arsenic also inhibited DNA ligases I and II, enzymes which play a role in DNA repair, with a sublinear dose response. In some cases, protective effects of relatively low exposures to arsenic have been observed, again consistent with sublinearity. We discuss recent theories on the mechanism of arsenic carcinogenicity and the potential implications for dose–response modeling and risk assessment. Overall, based on available arsenic genotoxicity data, we conclude that it is likely that arsenic indirectly induces genetic damage with a sublinear dose response in humans, thus providing a biological basis for a sublinear dose–response relationship for human cancer. Furthermore, these results suggest that linear dose–response modeling from populations experiencing high arsenic exposures is likely to overpredict cancer risks at relatively low arsenic levels.

Abstract  This working paper summarizes the known ultrastructural and biochemical effects of lead, mercury, cadmium, and arsenic on subcellular organelle systems following in vivo administration. Documented metal-induced alterations in nuclear, mitochondrial, microsomal, and lysosomal functions are discussed in relation to their potential impact on cellular responses to other environmental agents. Each of the above elements has been found to interfere with normal cellular replication and genetic processes. Mitochondrial swelling and depression of respiratory function are discussed in relation to known metal-specific perturbations of mitochondrial heme biosynthetic pathway enzymes. Inhibition of microsomal enzyme activities and protein synthesis by lead and mercury is compared to the apparent absence of such effects following arsenic or cadmium exposure. Lysosomal uptake of all the metals is documented, but biochemical alterations in these structures have been reported for only mercury and cadmium. It is concluded that these toxic metals are capable of interacting with, and biochemically altering major cellular systems at dose levels below those required to produce signs of overt metal toxicity. The impact of these effects on cellular response to other metals and xenobiotics in complex exposure situations is presently unknown, and further research is urgently needed in this area.

Abstract  Arsenic at a nonlethal level in drinking water consumed over a period of time has been reported to produce chronic toxicity and various types of health problems ranging from skin cancer to disturbance in memory. Neurotoxic effects have been reported in clinical cases with chronic exposure to arsenic. Physiological detoxication of arsenic occurs partially through methylation. Arsenic and its methylated derivatives are distributed in different organs and systems. The present study examined the possible interference in the neuronal development and differentiation due to the exposure to arsenic during gestation. The experiments were carried out to examine short and long term effects of arsenic on brain explants and cells grown and maintained in tissue culture system. The effects of arsenic exposure showed changes in brain cell membrane function indicated by generation and release of reactive oxygen-nitrogen intermediates. On the morphological aspect the explants' growth was reduced, ground matrix was lost and neural networking was inhibited. Cells showed signs of apoptotic changes. Arsenic toxicity may induce damage to brain cells prior to more visible clinical conditions. The deleterious effects also pass from the maternal to fetal tissue across the transplacental barrier.

Abstract  The U. S. Environmental Protection Agency’s (EPA) Office of Pesticide Programs (OPP), Office of Water (OW), and Office of Research and Development (ORD) coordinated the development of two scientific documents that address the carcinogenicity of Dimethylarsinic Acid (DMAV) and inorganic arsenic (iAs). In response to an Agency request, the Science Advisory Board (SAB) convened an expert panel to review and comment on key scientific issues presented in these two documents, including: (a) the metabolism and toxic responses of arsenic species; (b) mode(s) of carcinogenic action; (c) data selection for dose-response assessment; and (d) approaches and methods for low-dose extrapolation for DMAV and iAs. The SAB Panel supported the Agency’s conclusion that on the basis of available data, human exposure to DMAV appears to result in a narrower spectrum of active metabolites than those expected in the metabolic profile associated with exposure to iAs. Therefore, the Panel agreed with EPA that, in the absence of human data on DMAV, the bladder tumor data from DMAV rat bioassays is better suited for DMAV cancer risk assessment than is epidemiology data from iAs exposure. The Panel, however, noted that there remain significant uncertainties associated with the use of animal data for DMAV cancer risk assessment due to the observed metabolic differences between rats and humans. The Panel agreed with the Agency’s conclusion that DMAV-induced bladder cancer in rats, at high dose, is mediated by a cytotoxic mode of action, and that this MOA should be considered relevant to humans. However, the Panel concluded there are not sufficient data to support a reactive oxygenated species-mediated mode of direct genetic action for DMAV. The Panel supported the nonlinear approach for low dose extrapolation of DMAV and the use of uncertainty factors to account for interspecies differences and human variability for sensitive human populations, and concluded that presently there is no arsenic-specific information that can inform the choice of specific values. This means that, at least for now, such choices must be based on more general considerations, including EPA’s science policy judgment of the degree of precaution that it deems appropriate. EPA concluded that the mechanisms by which inorganic arsenic induces bladder cancer in humans are not yet known, but they are likely to be mediated by multiple modes of action. The Agency used a linear default approach for low dose extrapolation because it lacked a full understanding of the iAs modes of carcinogenic action. The Panel agreed that available human and animal data do not fully describe the shape of the iAs carcinogenic dose-response curve at low doses. Given the considerable uncertainties regarding low dose extrapolation, the Panel supported the use of a linear cancer risk model for iAs as recommended by the National Research Council in its 2001 report. The Panel also supported the use of the epidemiologic data on the Taiwanese population for estimating human cancer risk for iAs especially to identify the potential range of responses of human populations. However, the Panel recognized limitations to these data, and that there is some evidence on iAs from animal toxicology, pharmacokinetics, and pharmacodynamics research, that suggests other than a linear bladder cancer dose-response. The Panel urged the Agency to consider other epidemiologic studies from the U.S. and other countries, utilizing a uniformset of evaluative criteria. The Panel also recommended sensitivity analyses be conducted to account for human variability in drinking water consumption rates, dietary intake of iAs from food, and certain other assumptions currently used in EPA’s assessment. The Panel made several suggestions for improvements in the currently applied risk model’s programming and documentation conventions. Finally, the Panel believes there is a critical need for a continued research effort to strengthen EPA’s cancer risk assessment for DMAV and iAs. The scientific bases for the Panel’s conclusions and research recommendations are detailed throughout this report.

Abstract  Chronic arsenic exposure is associated with cardiovascular abnormalities. Prolongation of the QT (time between initial deflection of QRS complex to the end of T wave) interval and profound repolarization changes on electrocardiogram (ECG) have been reported in patients with acute promyelocytic leukemia treated with arsenic trioxide. This acquired form of long QT syndrome can result in life-threatening arrhythmias.

The objective of this study was to assess the cardiac effects of arsenic by investigating QT interval alterations in a human population chronically exposed to arsenic.

Residents in Ba Men, Inner Mongolia, have been chronically exposed to arsenic via consumption of water from artesian wells. A total of 313 Ba Men residents with the mean arsenic exposure of 15 years were divided into three arsenic exposure groups: low (< or = 21 microg/L), medium (100-300 microg/L), and high (430-690 microg/L). ECGs were obtained on all study subjects. The normal range for QTc (corrected QT) interval is 0.33-0.44 sec, and QTc > or = 0.45 sec was considered to be prolonged.

The prevalence rates of QT prolongation and water arsenic concentrations showed a dose-dependent relationship (p = 0.001). The prevalence rates of QTc prolongation were 3.9, 11.1, 20.6% for low, medium, and high arsenic exposure, respectively. QTc prolongation was also associated with sex (p < 0.0001) but not age (p = 0.486) or smoking (p = 0.1018). Females were more susceptible to QT prolongation than males.

We found significant association between chronic arsenic exposure and QT interval prolongation in a human population. QT interval may potentially be useful in the detection of early cardiac arsenic toxicity.

Abstract  To gain insight into plant responses to arsenic, the effect of arsenic exposure on maize (Zea mays L.) root proteome has been examined. Maize seedlings were fed hydroponically with 300 microM sodium arsenate or 250 microM sodium arsenite for 24 h, and changes in differentially displayed proteins were studied by two-dimensional electrophoresis and digital image analysis. About 10% of total detected maize root proteins (67 out of 700) were up- or down-regulated by arsenic, among which 20 were selected as being quite reproducibly affected by the metalloid. These were analyzed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry and 11 of them could be identified by comparing their peptide mass fingerprints against protein- and expressed sequence tag-databases. The set of identified maize root proteins highly responsive to arsenic exposure included a major and functionally homogeneous group of seven enzymes involved in cellular homeostasis for redox perturbation (e.g., three superoxide dismutases, two glutathione peroxidases, one peroxiredoxin, and one p-benzoquinone reductase) besides four additional, functionally heterogeneous, proteins (e.g., ATP synthase, succinyl-CoA synthetase, cytochrome P450 and guanine nucleotide-binding protein beta subunit). These findings strongly suggest that the induction of oxidative stress is a main process underlying arsenic toxicity in plants.

Abstract  Arsenic is a well-documented human carcinogen and is known to cause oxidative stress in cultured cells and animals. A hospital-based case-control study was conducted to evaluate the relationship among the levels of urinary 8-hydroxydeoxyguanosine (8-OHdG), the arsenic profile, and urothelial carcinoma (UC). Urinary 8-OHdG was measured by using high-sensitivity enzyme-linked immunosorbent assay (ELISA) kits. The urinary species of inorganic arsenic and their metabolites were analyzed by high-performance liquid chromatography (HPLC) and hydride generator-atomic absorption spectrometry (HG-AAS). This study showed that the mean urinary concentration of total arsenics was significantly higher, at 37.67±2.98 μg/g creatinine, for UC patients than for healthy controls of 21.10±0.79 μg/g creatinine (p<0.01). Urinary 8-OHdG levels correlated with urinary total arsenic concentrations (r=0.19, p<0.01). There were significantly higher 8-OHdG levels, of 7.48±0.97 ng/mg creatinine in UC patients, compared to healthy controls of 5.95±0.21 ng/mg creatinine. Furthermore, female UC patients had higher 8-OHdG levels of 9.22±0.75 than those of males at 5.76±0.25 ng/mg creatinine (p<0.01). Multiple linear regression analyses revealed that high urinary 8-OHdG levels were associated with increased total arsenic concentrations, inorganic arsenite, monomethylarsonic acid (MMA), and dimethylarsenate (DMA) as well as the primary methylation index (PMI) even after adjusting for age, gender, and UC status. The results suggest that oxidative DNA damage was associated with arsenic exposure, even at low urinary level of arsenic.

Abstract  The expression of beclin-1 in normal human bladder and in Cd(2+) and As(3+) exposed and transformed urothelial cells (UROtsa) was examined in this study. It was shown using a combination of real-time PCR, Western analysis and immunohistochemistry that beclin-1 was expressed in the urothelial cells of the normal bladder. It was also demonstrated that the parental UROtsa cell line expressed beclin-1 mRNA and protein at levels similar to that of the in situ urothelium. The level of beclin-1 expression underwent only modest alterations when the UROtsa cells were malignantly transformed by Cd(2+) or As(3+) or when the parental cells were exposed acutely to Cd(2+) or As(3+). While there were instances of significant alterations at individual time points and within cell line-to-cell line comparisons there was no evidence of a dose-response relationship or correlations to the phenotypic properties of the cell lines. Similar results were obtained for the expression of the Atg-5, Atg-7, Atg-12 and LC3B autophagy-related proteins. The findings provide initial evidence for beclin-1 expression in normal bladder and that large alterations in the expression of beclin-1 and associated proteins do not occur when human urothelial cells are malignantly transformed with, or exposed to, either Cd(2+) or As(3+.).

Abstract  Quite a few new thioarsenicals have recently been found in urine of arsenic-exposed humans and animals, and some of them have been shown to be highly toxic to cells. However, little is known about their toxic effects and metabolism in the body. In order to elucidate the toxic mechanism of thioarsenicals, we further focused on the distribution and metabolism of monomethylmonothioarsonic acid (MMMTA(V)) in rats. MMMTA(V) was synthesized chemically and injected intravenously into rats at the dose of 0.5mg As/kg, followed by speciation analysis of selected organs and body fluids at 10 min and 12h after the injection. MMMTA(V) was excreted into urine in its intact form, and approximately 35% of the dose was recovered in urine at 12h after the injection, suggesting that MMMTA(V) was taken up more effectively by organs/tissues than non-thiolated, monomethylarsonous acid (MMA(V)) previously studied. On the other hand, the liver and kidneys contained arsenic that was in a protein-binding form with free forms of DMA(V) or DMDTA(V) at 10 min, and disappeared at 12h after the injection. Moreover, these bound arsenic species in kidneys were converted back to MMA(V) after oxidation with H(2)O(2), suggesting that the arsenic bound to proteins had been reduced within the body and was in a trivalent oxidation state. In red blood cells (RBCs), most of the arsenic was in the form of DMA(III) bound to hemoglobin (Hb), and approximately 40% of the dose was recovered in RBCs at 12h after injection. These results indicate that arsenic accumulated preferentially in RBCs after being transformed to DMA(III). In addition, we have also discussed the effect of MMMTA(V) on viability of human bladder cancer T24 cells in comparison with MMA(V). Consequently, MMMTA(V) was assumed to be a more toxic arsenic metabolite than non-thiolated MMA(V).

Abstract  BACKGROUND: Inorganic arsenic is an environmental carcinogen that may act through multiple mechanisms including formation of methylated derivatives in vivo. Sodium arsenite (up to 5.0 microM) renders arsenic methylation-competent TRL1215 rat liver epithelial cells tumorigenic in nude mice at 18 weeks of exposure and arsenic methylation-deficient RWPE-1 human prostate epithelial cells tumorigenic at 30 weeks of exposure. We assessed the role of arsenic biomethylation in oxidative DNA damage (ODD) using a recently developed immuno-spin trapping method. METHODS: Immuno-spin trapping was used to measure ODD after chronic exposure of cultured TRL1215 vs RWPE-1 cells, or of methylation-competent UROtsa/F35 vs methylation-deficient UROtsa human urothelial cells, to sodium arsenite. Secreted matrix metalloproteinase (MMP)-2 and -9 activity, as analyzed by zymography, cellular invasiveness by using a transwell assay, and colony formation by using soft agar assay were compared in cells exposed to arsenite with and without selenite, an arsenic biomethylation inhibitor, to assess the role of ODD in the transition to an in vitro cancer phenotype. RESULTS: Exposure of methylation-competent TRL1215 cells to up to 1.0 microM sodium arsenite was followed by a substantial increase in ODD at 5-18 weeks (eg, at 16 weeks with 1.0 microM arsenite, 1138% of control, 95% confidence interval [CI] = 797% to 1481%), whereas exposure of methylation-deficient RWPE-1 cells to up to 5.0 microM arsenite did not increase ODD for a 30-week period. Inhibition of arsenic biomethylation with sodium selenite abolished arsenic-induced ODD and invasiveness, colony formation, and MMP-2 and -9 hypersecretion in TRL1215 cells. Arsenic induced ODD in methylation-competent UROtsa/F35 cells (eg, at 16 weeks, with 1.0 microM arsenite 225% of control, 95% CI = 188% to 262%) but not in arsenic methylation-deficient UROtsa cells, and ODD levels corresponded to the levels of increased invasiveness, colony formation, and hypersecretion of active MMP-2 and -9 seen after transformation to an in vitro cancer phenotype. CONCLUSION: Arsenic biomethylation appears to be obligatory for arsenic-induced ODD and appears linked in some cells with the accelerated transition to an in vitro cancer phenotype.

Abstract  Arsenic, a curative agent for acute promyelocytic leukemia, induces cell apoptosis and degradation of BCR-ABL in chronic myelogenous leukemia (CML). We demonstrated that ubiquitination and degradation of BCR-ABL was mediated by c-CBL, a RING-type E3 ligase that was also shown to be involved in ubiquitination for many other receptor/protein tyrosine kinases. Our data showed that c-CBL protein was considerably up-regulated by arsenic sulfide (As4S4). Interestingly, arsenic directly bound the RING finger domain of c-CBL to inhibit its self-ubiquitination/degradation without interfering with the enhancement of ubiquitination and subsequent proteolysis of its substrate BCR-ABL. Degradation of BCR-ABL due to c-CBL induction as a result of arsenic treatment was also observed in vivo in CML mice. These findings provide insight into the molecular mechanisms of arsenic and further support its therapeutic applications in CML in combination with tyrosine kinase inhibitors and potentially also in other malignancies involving aberrant receptor/protein tyrosine kinase signaling.

Abstract  Liver aquaglyceroporin AQP9 facilitates movement of trivalent inorganic arsenite (As(III)) and organic monomethylarsonous acid (MAs(III)). However, the transport pathway for the two major pentavalent arsenic cellular metabolites, MAs(V) and DMAs(V), remains unknown in mammals. These products of arsenic metabolism, in particular DMAs(V), are the major arsenicals excreted in the urine of mammals. In this study, we examined the uptake of the two pentavalent organic arsenicals by human AQP9 in Xenopus laevis oocytes. Xenopus laevis oocytes microinjected with AQP9 cRNA exhibited uptake of both MAs(V) and DMAs(V) in a pH-dependent manner. The rate of transport was much higher at acidic pH (pH5.5) than at neutral pH. Hg(II), an aquaporin inhibitor, inhibited transport of As(III), MAs(III), MAs(V) and DMAs(V) via AQP9. However, phloretin, which inhibits water and glycerol permeation via AQP9, can only inhibit transport of pentavalent MAs(V) and DMAs(V) but not trivalent As(III) and MAs(III), indicating the translocation mechanisms of these arsenic species are not exactly the same. Reagents such as FCCP, valinomycin and nigericin that dissipate transmembrane proton potential or change the transmemebrane pH gradient did not significantly inhibit all arsenic transport via AQP9, suggesting the transport of pentavalent arsenic is not proton coupled. The results suggest that in addition to the initial uptake of trivalent inorganic As(III) inside cells, AQP9 plays a dual role in the detoxification of arsenic metabolites by facilitating efflux from cells.

Abstract  There is growing evidence that chronic exposure of humans to inorganic arsenic, a potent environmental oxidative stressor, is associated with the incidence of type 2 diabetes (T2D). One critical feature of T2D is insulin resistance in peripheral tissues, especially in mature adipocytes, the hallmark of which is decreased insulin-stimulated glucose uptake (ISGU). Despite the deleterious effects of reactive oxygen species (ROS), they have been recognized as a second messenger serving an intracellular signaling role for insulin action. Nuclear factor erythroid 2-related factor 2 (NRF2) is a central transcription factor regulating cellular adaptive response to oxidative stress. This study proposes that in response to arsenic exposure, the NRF2-mediated adaptive induction of endogenous antioxidant enzymes blunts insulin-stimulated ROS signaling and thus impairs ISGU. Exposure of differentiated 3T3-L1 cells to low-level (up to 2μM) inorganic arsenite (iAs(3+)) led to decreased ISGU in a dose- and time-dependent manner. Concomitant to the impairment of ISGU, iAs(3+) exposure significantly attenuated insulin-stimulated intracellular ROS accumulation and AKT S473 phosphorylation, which could be attributed to the activation of NRF2 and induction of a battery of endogenous antioxidant enzymes. In addition, prolonged iAs(3+) exposure of 3T3-L1 adipocytes resulted in significant induction of inflammatory response genes and decreased expression of adipogenic genes and glucose transporter type 4 (GLUT4), suggesting chronic inflammation and reduction in GLUT4 expression may also be involved in arsenic-induced insulin resistance in adipocytes. Taken together our studies suggest that prolonged low-level iAs(3+) exposure activates the cellular adaptive oxidative stress response, which impairs insulin-stimulated ROS signaling that is involved in ISGU, and thus causes insulin resistance in adipocytes.