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Differential expression of molecular markers in arsenic- and non-arsenic-related urothelial cancer.

BACKGROUND: Little is known about the mechanisms of arsenic-related urothelial cancer (AsUC). The aim of this study was to reveal the differential expression of molecular markers between AsUC and non-arsenic-related UC (non-AsUC). MATERIALS AND METHODS: Tissues of AsUC (n=33), non-AsUC (n=20) and normal bladder urothelia from patients with benign diseases (n=4) were examined for multiple selected molecular markers responsible for various cellular functions, includingglutathione, GST-pi, Bcl-2, p53 and c-Fos. RESULTS: The mean cellular glutathione content of normal mucosal samples (33.4 +/- 7.2 microM/mg protein) was significantly higher than either non-AsUC (22.8 +/- 1.8, p = 0.04) or AsUC (16.4 +/- 1.6, p = 0.002). The glutathione content of non-AsUC was higher than that of AsUC (p = 0.012). The expressions of Bcl-2 and c-Fos in AsUC were significantly higher than those in non-AsUC (p = 0.004 and p = 0.02, respectively). CONCLUSION: The carcinogenic pathway for AsUC is different, in part, from that of non-AsUC. Cellular glutathione contents may be down-regulated during urothelial carcinogenesis. Bcl-2 and c-Fos may play important roles in arsenic-mediated carcinogenesis of the urothelium.

Arsenic Poisoning↗

[Arsenic poisoning through the roof. A case of arsenic poisoning in cattle].

A case of arsenic poisoning in cattle is reported, in which six out of fifteen animals died. The source of poisoning was traced to a substance in the loft of the hay-barn, which probably consisted of the remains of a pesticide which had been there for several years. This had become liquefied as the result of a leak in the roof and had found its way into the hay. The arsenic content of the substance ranged from 17 to 37 per cent, that of the hay being up to 0.3 per cent. The concentration of arsenic in the organs, blood, faeces and urine of the animals were indicative of poisoning. The main clinical findings consisted in profuse diarrhoea, loss of appetite, muscular weakness and an ataxic gait. Dehydration set in later and the animals became deeply soporous. Symptomatic treatment was mainly directed against dehydration, hypoglycaemia and hypocalcaemia. The greater part of the animals gradually recovered. Marked hyperaemia of the abomasal mucosa and, to a less extent, hyperaemia of the small and large intestines was observed on post-mortem examination.

Animal Feed↗

Enzymatic methylation of arsenic species and other new approaches to arsenic toxicity.

Arsenic metabolism has typically been studied by administering arsenate or arsenite into animals and humans and then studying the metabolites excreted in the urine. Although such studies have yielded information about the beginning and the end of the metabolic pathways for the metabolism of inorganic arsenic compounds, any statements as to the molecular mechanisms of these reactions have had to be highly speculative. Now that the rabbit and the rhesus monkey liver enzymes that transfer methyl groups from S-adenosylmethionine to arsenite and monomethlyarsonic acid have been purified and the reactions characterized, meaningful investigations of species diversity and polymorphism of these enzymes have become possible. New World animals studied thus far appear to be deficient in or totally lacking these enzymes. Old World animals, with the exception of the chimpanzee, have ample amounts of arsenite and monomethylarsonic acid methyltransferases. A hypothesis that the lack of arsenite methyltransferases may have had an evolutionary advantage for certain species is proposed.

Animals↗

Effects of administration styles of arsenic trioxide on intracellular arsenic concentration, cell differentiation and apoptosis.

We studied the effects of varying and steady-state concentrations of arsenic trioxide (As2O3) on apoptosis and differentiation of several cell lines in vitro. We also studied the same effects of fluctuating vs constant concentrations of As2O3 in vivo in patients treated with daily 3-hour fast infusions or daily slow, continuous infusions. Intracellular concentrations of arsenic and apoptosis rate were higher whereas differentiation was reduced in cells exposed to constant concentrations of As2O3.

Apoptosis↗

Controlling the oxidation state of arsenic in cyclic arsenic cations.

Reduction of AsCl3 with SnCl2, followed by treatment of the "AsCl" with a 1,4-diimine results in electron transfer and formation of an arsenic(III) salt, while treatment of this arsenic(I) reagent or AsI3 with an alpha,alpha'-diiminopyridine ligand forms an arsenic(I) salt.

Journal Article↗

Blood arsenic as a biomarker of arsenic exposure: results from a prospective study.

Exposure to arsenic (As)-contaminated drinking water affects millions of people worldwide. Arsenic exposure is associated with skin lesions, skin, lung, kidney and liver cancers, neurologic and cardiovascular effects. Past studies involving biomarkers of As exposure have typically examined urinary As (UAs) (adjusted for urinary creatinine), hair or toenail As, but not blood As (BAs) since blood concentrations are exceedingly low and are not detectable by conventional atomic absorption spectrophotometric techniques. In a case-cohort analysis of 303 newly diagnosed cases of skin lesions, and 849 subcohort members randomly selected from 8092 participants in the health effects of as longitudinal study (HEALS) in Araihazar, Bangladesh, we measured blood, urine and water As concentrations, and examined their associations with each other, and with the risk for skin lesions. BAs concentrations were highly correlated with creatinine-adjusted UAs concentrations (r=0.85) and with water As (WAs) (r=0.75). We observed consistent dose-response relationships between the risk of skin lesions and all the measures of As exposure. Rate ratios (RRs) for skin lesions by quintile of As exposure, adjusted for age and gender, revealed that the two highest quintiles were significantly related to an increased risk of skin lesions for each measure of exposure: BAs, UAs, WAs and a time-weighted water As variable. This prospective study confirms the increased risk of skin lesions in relation to As concentrations in blood, urine and water and also establishes that BAs is a useful biomarker of As exposure in this study population.

Adult↗

Determination of arsenic in urine by atomic absorption spectrophotometry for biological monitoring of occupational exposure to arsenic.

An atomic absorption spectrophotometric (AAS) method was successfully applied to analysis of urine for arsenic (As) as a measure of biological monitoring of occupational exposure to As in Vietnam. The application of the method to urine samples from 75 non-exposed control urbanites (after 2-day abstinence from sea foods) gave a reference level of 62.4 +/- 11.6 microg/l (as mean +/- S.D.), from which the upper limit of the normal value (74 microg/l as mean +/- 1 S.D.) and the acceptable limit (100 microg/l as mean +/- 3S.D.) were deduced. Further application to urine samples from 147 workers occupationally exposed to As in Bacthai Non-ferrous Metallurgic Corporation showed significantly elevated levels of As in urine, with mean +/- S.D. of 78.5 +/- 20.2 microg/l. Improvement of working conditions to reduce As exposure resulted in substantial reduction in the ratio of those with urinary As at the level in excess of the acceptable limit. The practical importance of total arsenic determination in urine after 2-day sea food abstinence is discussed in connection with current conditions in analytical laboratories in Vietnam.

Adult↗

Acute arsenic intoxication from environmental arsenic exposure.

Reports of acute arsenic poisoning arising from environmental exposure are rare. Two cases of acute arsenic intoxication resulting from ingestion of contaminated well water are described. These patients experienced a variety of problems: acute gastrointestinal symptoms, central and peripheral neurotoxicity, bone marrow suppression, hepatic toxicity, and mild mucous membrane and cutaneous changes. Although located adjacent to an abandoned mine, the well water had been tested for microorganisms only and was found to be "safe." Regulations for testing of water from private wells for fitness to drink are frequently nonexistent, or only mandate biologic tests for microorganisms. Well water, particularly in areas near mining activity, should be tested for metals.

Acute Disease↗

Urinary trivalent methylated arsenic species in a population chronically exposed to inorganic arsenic.

Chronic exposure to inorganic arsenic (iAs) has been associated with increased risk of various forms of cancer and of noncancerous diseases. Metabolic conversions of iAs that yield highly toxic and genotoxic methylarsonite (MAsIII) and dimethylarsinite (DMAsIII) may play a significant role in determining the extent and character of toxic and cancer-promoting effects of iAs exposure. In this study we examined the relationship between urinary profiles of MAsIII and DMAsIII and skin lesion markers of iAs toxicity in individuals exposed to iAs in drinking water. The study subjects were recruited among the residents of an endemic region of central Mexico. Drinking-water reservoirs in this region are heavily contaminated with iAs. Previous studies carried out in the local populations have found an increased incidence of pathologies, primarily skin lesions, that are characteristic of arseniasis. The goal of this study was to investigate the urinary profiles for the trivalent and pentavalent As metabolites in both high- and low-iAs-exposed subjects. Notably, methylated trivalent arsenicals were detected in 98% of analyzed urine samples. On average, the major metabolite, DMAsIII, represented 49% of total urinary As, followed by DMAsV (23.7%), iAsV (8.6%), iAsIII (8.5%), MAsIII (7.4%), and MAsV (2.8%). More important, the average MAsIII concentration was significantly higher in the urine of exposed individuals with skin lesions compared with those who drank iAs-contaminated water but had no skin lesions. These data suggest that urinary levels of MAsIII, the most toxic species among identified metabolites of iAs, may serve as an indicator to identify individuals with increased susceptibility to toxic and cancer-promoting effects of arseniasis.

Adolescent↗

Combined arsenic and retinoic acid treatment enhances differentiation and apoptosis in arsenic-resistant NB4 cells.

In the acute promyelocytic leukemia (APL) cell line NB4, as well as in APL patients' cells, arsenic trioxide (As2O3) leads to incomplete cell maturation, induction of apoptosis, as well as to the degradation of the oncogenic PML/RARalpha fusion protein. We have isolated an arsenic-resistant NB4 subline (NB4-AsR), which fails to undergo apoptosis, but maintains the partial differentiation response to this drug. When grown in the presence of As2O3, NB4-AsR cells degrade PML/RARalpha, slightly differentiate, and become more sensitive to serum deprivation-induced apoptosis. Similarly, in RA-resistant NB4-R1 cells, RA induced a significant PML/RARalpha degradation and yet failed to induce cell maturation. Thus, As2O3- or retinoic acid (RA)-induced PML/RARalpha degradation may be a prerequisite, but is not sufficient for the full differentiative/apoptotic response to these drugs. Strikingly, RA-triggered differentiation and apoptosis were greatly accelerated in As2O3-treated NB4-AsR cells. The synergism between these two agents in this setting could provide an experimental basis for combined or sequential RA/As2O3 therapies.

Antineoplastic Agents↗

Diastereoisomerism of thiol complexes of arsenic acids and pseudoasymmetry of arsenic: a 1H and 13C NMR study.

Diastereoisomeric complexes of methylphenylarsinic acid and (L)-glutathione could be partially separated by HPLC, but the separated compounds rapidly racemized, presumably by pyramidal inversion at the arsenic atom. Hydrolysis of the diastereoisomeric complexes yielded methylphenylarsinous acid as a pair of enantiomers revealed by a 1H NMR study with an asymmetric lanthanide shift reagent. Methylphenylarsinous acid was also synthesized as an enantiomeric pair, shown by an asymmetric shift reagent experiment, by the hydrolysis of iodomethylphenylarsine. 1H and 13C NMR spectroscopy were used to demonstrate that complexing of phenylarsonic acid with (R,S)-3-mercapto-1,2-propanediol and with (R,S)-1-mercapto-2-propanol yielded, in each case, a pair of enantiomers, PhAs[(R)-ligand)]2, PhAs[(S)-ligand)]2, in which the homomorphic ligands were diastereotopic, and a pair of diastereoisomers, PhAs[(R)-ligand][(S)-ligand], which differed from each other in the configuration about the pseudoasymmetric arsenic atom.

Arsenates↗

Metabolic fate and cardiovascular effects of arsenic in rats and rabbits chronically exposed to trivalent and pentavalent arsenic.

Male rats and female rabbits exposed for eighteen and ten months, respectively, to 50 micrograms/ml of trivalent arsenic (As III) in drinking water showed reduction of stroke volume (SV) and cardiac output (CO) and increase of vascular resistance (VR), while no cardiovascular change was observed in male rats receiving 50 micrograms/ml of pentavalent arsenic (As V) for eighteen months. The exposed animals showed specific alterations in some neurohumoral and effector mechanisms regulating cardiovascular function (CF). Elevated levels of urinary As were found in all treated groups. However, there were significant differences in the urinary levels of dimethylarsenic acid (DMA), monomethylarsenic acid (MMA) and inorganic As, which depended on both animal species and the valency state of As. Thus, As(III) was methylated to a greater extent than As(V) in rats, with less elimination of inorganic As, while rabbits had a lower capacity to methylate As and to retain As in tissues than rats. Either cardiovascular effects or metabolism of As seemed to depend on its valency state and animal species.

Animals↗

Antioxidative responses to arsenic in the arsenic-hyperaccumulator Chinese brake fern (Pteris vittata L.).

This study measured antioxidative responses of Chinese brake fern (Pteris vittata L.) upon exposure to arsenic (As) of different concentrations. Chinese brake fern was grown in an artificially-contaminated soil containing 0 to 200 mg As kg(-1) (Na2HAsO4) for 12 weeks in a greenhouse. Soil As concentrations at < or =20 mg kg(-1) enhanced plant growth, with 12-71% biomass increase compared to the control. Such beneficial effects were not observed at >20 mg As kg(-1). Plant As concentrations increased with soil As concentrations, with more As being accumulated in the fronds (aboveground biomass) than in the roots and with maximum frond As concentration being 4675 mg kg(-1). Arsenic uptake by Chinese brake enhanced uptake of nutrient elements K, P, Fe, Mn, and Zn except Ca and Mg, whose concentrations mostly decreased. The contents of non-enzymatic antioxidants (glutathione, acid-soluble thiol) followed similar trends as plant As concentrations, increasing with soil As concentrations, with greater contents in the fronds than in the roots especially when exposed to high As concentrations (>50 mg kg(-1)). The activities of enzymatic antioxidants (superoxide dismutase, catalase, ascorbate peroxidase, guaiacol peroxidase) in Chinese brake followed the same trends as plant biomass, increasing with soil As up to 20 mg kg(-1) and then decreased. The results indicated though both enzymatic and non-enzymatic antioxidants played significant roles in As detoxification and hyperaccumulation in Chinese brake, the former is more important at low As exposure (< or =20 mg kg(-1)), whereas the latter is more critical at high As exposure (50-200 mg kg(-1)).

Antioxidants↗

Effects of arsenic on concentration and distribution of nutrients in the fronds of the arsenic hyperaccumulator Pteris vittata L.

Pteris vittata was the first terrestrial plant known to hyperaccumulate arsenic (As). However, it is unclear how As hyperaccumulation influences nutrient uptake by this plant. P. vittata fern was grown in soil spiked with 0-500 mg As kg(-1) in the greenhouse for 24 weeks. The concentrations of essential macro- (P, K, Ca, and Mg) and micro- (Fe, Mn, Cu, Zn, B and Mo) elements in the fronds of different age were examined. Both macro- and micronutrients in the fronds were found to be within the normal concentration ranges for non-hyperaccumulators. However, As hyperaccumulation did influence the elemental distribution among fronds of different age of P. vittata. Arsenic-induced P and K enhancements in the fronds contributed to the As-induced growth stimulation at low As levels. The frond P/As molar ratios of 1.0 can be used as the threshold value for normal growth of P. vittata. Potassium may function as a counter-cation for As in the fronds as shown by the As-induced K increases in the fronds. The present findings not only demonstrate that P. vittata has the ability to maintain adequate concentrations of essential nutrients while hyperaccumulating As from the soil, but also have implications for soil management (fertilization in particular) of P. vittata in As phytoextraction practice.

Arsenic↗

Total arsenic, inorganic arsenic, lead and cadmium contents in edible seaweed sold in Spain.

Total arsenic, inorganic arsenic, lead and cadmium contents were determined in 112 samples of seaweed preparations sold in Spain (seaweed packed in plastic or cardboard box, seaweed in the form of tablets and concentrates, foods containing seaweed, and canned seaweed). The concentration ranges found, expressed in mg/kg, dry weight, were: total As (0.031-149), inorganic As (<0.014-117), Pb (<0.050-12.1) and Cd (<0.003-3.55). For all the contaminants there were failures to comply with legislated values. In particular, all the samples of Hizikia fusiforme exceeded the inorganic As limit established in some countries, and a considerable number of species exceeded the Cd limit set by international regulations. With respect to food safety, consumption of 3 g/day of the samples analysed could represent up to 15% of the respective Tolerable Daily Intakes (TDI) established by the WHO. The situation is especially alarming for intake of inorganic As from H. fusiforme, which can be three times the TDI established.

Arsenic↗

Metabolism and toxicity of arsenic in human urothelial cells expressing rat arsenic (+3 oxidation state)-methyltransferase.

The enzymatic methylation of inorganic As (iAs) is catalyzed by As(+3 oxidation state)-methyltransferase (AS3MT). AS3MT is expressed in rat liver and in human hepatocytes. However, AS3MT is not expressed in UROtsa, human urothelial cells that do not methylate iAs. Thus, UROtsa cells are an ideal null background in which the role of iAs methylation in modulation of toxic and cancer-promoting effects of this metalloid can be examined. A retroviral gene delivery system was used in this study to create a clonal UROtsa cell line (UROtsa/F35) that expresses rat AS3MT. Here, we characterize the metabolism and cytotoxicity of arsenite (iAs(III)) and methylated trivalent arsenicals in parental cells and clonal cells expressing AS3MT. In contrast to parental cells, UROtsa/F35 cells effectively methylated iAs(III), yielding methylarsenic (MAs) and dimethylarsenic (DMAs) containing either As(III) or As(V). When exposed to MAs(III), UROtsa/F35 cells produced DMAs(III) and DMAs(V). MAs(III) and DMAs(III) were more cytotoxic than iAs(III) in UROtsa and UROtsa/F35 cells. The greater cytotoxicity of MAs(III) or DMAs(III) than of iAs(III) was associated with greater cellular uptake and retention of each methylated trivalent arsenical. Notably, UROtsa/F35 cells were more sensitive than parental cells to the cytotoxic effects of iAs(III) but were more resistant to cytotoxicity of MAs(III). The increased sensitivity of UROtsa/F35 cells to iAs(III) was associated with inhibition of DMAs production and intracellular accumulation of MAs. The resistance of UROtsa/F35 cells to moderate concentrations of MAs(III) was linked to its rapid conversion to DMAs and efflux of DMAs. However, concentrations of MAs(III) that inhibited DMAs production by UROtsa/F35 cells were equally toxic for parental and clonal cell lines. Thus, the production and accumulation of MAs(III) is a key factor contributing to the toxicity of acute iAs exposures in methylating cells.

Animals↗

Studies on the accumulation and transformation of arsenic in freshwater organisms II. Accumulation and transformation of arsenic compounds by Tilapia mossambica.

Bioaccumulation and biotransformation of arsenic (As) compounds in freshwater Tilapia mossambica was investigated. The direct accumulation of As by T mossambica was proportional to the concentration of arsenicals in water. Small amounts of accumulated As were transformed to methylated As, including trimethylarsenic (TMA) species. Accumulation and transformation of As(III) by T. mossambica via freshwater food chain results in the transformation of As(III) to As(V) with little biomethylation of accumulated As. Approximately 90% of accumulated As was depurated to water.

Animals↗

Validated determination of total arsenic species of toxicological interest (arsenite, arsenate and their metabolites) by atomic absorption spectrometry after separation from dietary arsenic by liquid extraction: toxicological applications.

A validated method for the selective extraction of total As species of toxicological interest (arsenite, arsenate and mono- and dimethylated arsenic species) from urine, followed by atomic absorption spectrometric determination, is described. The mechanisms involved in extraction were studied and the extraction method was optimized. The urine sample was acidified with concentrated HCl and KI and sodium hypophosphite were added. Under these conditions, As species were reduced to their corresponding iodide arsines, extracted with toluene and back-extracted with 1 mmol l-1 NaOH solution. Only inorganic arsenic and its metabolites in humans (monomethylarsonic and dimethylarsinic acid) were extracted. Arsenobetaine of dietary origin was not extracted. This method can detect if any As increase in urine originates from inorganic As intoxication or only from dietary non-toxic As species such as arsenobetaine.

Arsenates↗