Chronic low-level arsenic exposure causes gender-specific alterations in locomotor activity, dopaminergic systems, and thioredoxin expression in mice.

Arsenic (As) is a toxic metalloid widely present in the environment. Human exposure to As has been associated with the development of skin and internal organ cancers and cardiovascular disorders, among other diseases. A few studies report decreases in intelligence quotient (IQ), and sensory and motor alterations after chronic As exposure in humans. On the other hand, studies of rodents exposed to high doses of As have found alterations in locomotor activity, brain neurochemistry, behavioral tasks, and oxidative stress. In the present study both male and female C57Bl/6J mice were exposed to environmentally relevant doses of As such as 0.05, 0.5, 5.0, or 50 mg As/L of drinking water for 4 months, and locomotor activity was assessed every month. Male mice presented hyperactivity in the group exposed to 0.5 mg As/L and hypoactivity in the group exposed to 50 mg As/L after 4 months of As exposure, whereas female mice exposed to 0.05, 0.5, and 5.0 mg As/L exhibited hyperactivity in every monthly test during As exposure. Furthermore, striatal and hypothalamic dopamine content was decreased only in female mice. Also decreases in tyrosine hydroxylase (TH) and cytosolic thioredoxin (Trx-1) mRNA expression in striatum and nucleus accumbens were observed in male and female mice, respectively. These results indicate that chronic As exposure leads to gender-dependent alterations in dopaminergic markers and spontaneous locomotor activity, and down-regulation of the antioxidant capacity of the brain.

Assessment of exposure to mixture pollutants in Mexican indigenous children.

The aim of the present work was to complete an exposure assessment in three Mexican indigenous communities using the community-based health risk assessment, which is the first step in the CHILD framework. We used 1-hydroxypyrene (1-OHP) as an exposure biomarker to polycyclic aromatic hydrocarbons (PAHs) and trans, trans-muconic acid (t,t-MA) as an exposure biomarker to benzene, persistent organic pollutants (POPs), lead, manganese, arsenic, and fluoride. Anthropometric measurements were also taken. In these communities, high percentages of children with chronic malnutrition were found (28 to 49 %) based on their weight and age. All communities showed a high percentage of children with detectable levels of four or more compounds (70 to 82 %). Additionally, our results showed that in indigenous communities, children are exposed to elevated levels of certain environmental pollutants, including manganese with 17.6, 16.8, and 7.3 µg/L from SMP, TOC, and CUA, respectively. Lead and HCB levels were similar in the indigenous communities (2.5, 3.1, and 4.2 µg/dL and 2.5, 3.1, and 3.7 ng/mL, respectively). 1-OHP and t,t-MA levels were higher in TOC (0.8 µmol/mol of creatinine, 476 µg/g of creatinine, respectively) when compared with SMP (0.1 µmol/mol of creatinine, 215.5 µg/g of creatinine, respectively) and CUA (0.1 µmol/mol of creatinine, 185.2 µg/g of creatinine, respectively). DDE levels were 30.7, 26.9, and 9.6 ng/mL in CUA, SMP, and TOC, respectively. The strength of this study is that it assesses exposure to pollutants with indications for the resultant risk before an intervention is made by the CHILD program to manage this risk in the indigenous communities. Considering the large number of people, especially children, exposed to multiple pollutants, it is important to design effective intervention programs that reduce exposure and the resultant risk in the numerous indigenous communities in Mexico.

Arsenic increased lipid peroxidation in rat tissues by a mechanism independent of glutathione levels.

The role of lipid peroxidation in the mechanism of arsenic toxicity was investigated in female rats pretreated with N-acetylcysteine (NAC, a glutathione [GSH] inducer) or with buthionine sulfoximine (BSO, a GSH depletor). Rats were challenged with sodium arsenite, and sacrificed 1 hr after this treatment. Results showed that arsenic decreased GSH levels and increased lipid peroxidation in liver, kidney, and heart, with a larger effect at 18.2 mg/kg than at 14.8 mg/kg for lipid peroxidation induction. In the liver of rats treated with arsenic, pretreatment with NAC increased the levels of GSH and decreased lipid peroxidation. In kidney and heart, NAC pretreatment protected the tissues against arsenic-induced depletion of GSH levels, but the same degree of protection was not found for lipid peroxidation induction. In its turn, BSO had an additive effect with arsenic in lowering the levels of GSH in the liver and kidney, but an inverse correlation between GSH levels and lipid peroxidation was found only in liver. Arsenic content in tissues of rats pretreated with NAC was lower than in rats treated only with arsenic. In rats with depleted levels of GSH (BSO-pretreated rats), a shift in arsenic tissue distribution was found, with higher levels in skin and lower levels in kidney. A clear tendency for a positive correlation between arsenic concentration and lipid peroxidation levels was found in liver, kidney, and heart.

Effects of oral exposure to mining waste on in vivo dopamine release from rat striatum.

Several single components of mining waste (arsenic, manganese, lead, cadmium) to which humans are exposed at the mining area of Villa de la Paz, Mexico, are known to provoke alterations of striatal dopaminergic parameters. In this study we used an animal model to examine neurochemical changes resulting from exposure to a metal mixture. We used microdialysis to compare in vivo dopamine release from adult rats subchronically exposed to a mining waste by oral route with those from a control group and from a sodium arsenite group (25 mg/kg/day). We found that arsenic and manganese do accumulate in rat brain after 2 weeks of oral exposure. The mining waste group showed significantly decreased basal levels of dihydroxyphenylacetic acid (DOPAC; 66.7 +/- 7.53 pg/ microl) when compared to a control group (113.7 +/- 14.3 pg/ microl). Although basal dopamine release rates were comparable among groups, when the system was challenged with a long-standing depolarization through high-potassium perfusion, animals exposed to mining waste were not able to sustain an increased dopamine release in response to depolarization (mining waste group 5.5 +/- 0.5 pg/ microl versus control group 21.7 +/- 5.8 pg/ microl). Also, DOPAC and homovanillic acid levels were significantly lower in exposed animals than in controls during stimulation with high potassium. The arsenite group showed a similar tendency to that from the mining waste group. In vivo microdialysis provides relevant data about the effects of a chemical mixture. Our results indicate that this mining waste may represent a health risk for the exposed population.

The effects of sodium arsenite exposure on behavioral parameters in the rat.

Arsenic is a metalloid widely present in the environment. It is found in well water, soil, and air, and is also released from mining residues and industrial debris, among other anthropogenic sources. It has been previously reported that the content of catecholamines in striatum, hippocampus, and other cerebral regions changes in mice and rats exposed to arsenic. Few studies have examined behavioral alterations after intoxication with arsenic, and both increased and decreased locomotor activity, as well as learning deficits, have been described. In order to characterize the behavioral alterations induced by arsenic exposure, we exposed adult male Sprague-Dawley rats to 5, 10, and 20 mg/kg of arsenic by intragastric route for 2 or 4 weeks. Exposed rats showed reduced locomotor activity, which returned to control levels at the end of the intoxication period. We also found an increase in the number of errors in an egocentric task, alterations in monoamine content in midbrain and cortex, and increases in arsenic brain concentration, which were related to time of the exposure but not dose. These results indicate that short-term arsenic exposure induces neural and behavioral changes that may reflect a neurotoxic effect, and that these alterations are correlated to dose, time of exposure, and experimental conditions.

Arsenic-cadmium interaction in rats: toxic effects in the heart and tissue metal shifts.

Previously, we had shown that arsenic interacts with cadmium in rats; our results showed that the toxicity of a mixture of arsenic + cadmium cannot be predicted by the toxic mechanisms of the individual components. In this paper, we present further evidence about the interaction of arsenic and cadmium in rats. The results were: arsenic modified the 24 h-LD50 value of cadmium more clearly than cadmium did with the one of arsenic; based on the LD50 values, the mixtures we studied were more toxic than either metal alone. With single doses (As 10 mg/kg, Cd 2.6 mg/kg, and As 10 mg/kg + Cd 2.6 mg/kg) the mixture As + Cd was more toxic than each metal. At these doses, cadmium significantly induces the levels of glutathione, metallothionein, and lipid peroxidation in heart tissue, as compared to a saline group of rats. Arsenic incremented glutathione and lipid peroxidation at higher values than those obtained with cadmium. The mixture of As + Cd behaved as arsenic in the induction of lipid peroxidation and glutathione and like cadmium in metallothionein induction. Finally, rats treated with As + Cd had less Cd in liver than animals treated only with cadmium, and more As in heart tissue than rats treated only with arsenic. Our results give further evidence about the arsenic-cadmium interaction in rats, demonstrate the utility of employing different biomarkers in the study of chemical mixtures and indicate that heart tissue is affected not only by the mixture of As + Cd, but also by either metal alone.

Arsenic-cadmium interaction in rats.

Simultaneous exposure to cadmium and arsenic is highly probable in the urban area of San Luis Potosi, Mexico due to common localization of copper and zinc smelters. Therefore, in this work, rats were intraperitoneally exposed either to cadmium or arsenic alone, or simultaneously to both metals. The effects of these treatments on three different toxicological parameters were studied. Cadmium modified the LD50 of arsenic and conversely arsenic modified the LD50 for cadmium. At the histopathological level, arsenic appeared to protect against the cadmium effects, especially on testes. This protective effect seemed to be related to the glutathione levels found in this tissue: rats exposed to both arsenic and cadmium, presented glutathione values intermediate to those observed after exposure to either metal alone; arsenic had the highest value and cadmium the lowest. In liver, rats exposed to arsenic, cadmium or arsenic and cadmium, presented glutathione values below those in the saline group, with the lowest value corresponding to the arsenic and cadmium treatment. The results appear to support the proposed interaction between arsenic and cadmium and coexposure to both metals seems to alter certain effects produced by either metal alone.

Effects of arsenite on central monoamines and plasmatic levels of adrenocorticotropic hormone (ACTH) in mice.

We studied the effects of chronic arsenic exposure on brain monoamines and plasma levels of adrenocorticotropic hormone (ACTH) of mice. After weaning, mice received arsenic (0, 20, 40, 60 or 100 ppm) in drinking water over a period of 9 weeks. Monoamine content was quantified in different brain regions, arsenic was quantified in brain tissue and ACTH levels in plasma. Brain arsenic concentrations up to 200 ng/g showed a significant correlation with exposure levels and produced slight modifications in regional monoamine levels. ACTH plasma levels were significantly associated with norepinephrine (NE) concentrations in the medulla and pons, but not with hypothalamic NE levels. ACTH levels were significantly higher in the group exposed to 20 ppm. Dopamine showed significant dose-related decreases in the hypothalamus. These results show that chronic sodium arsenite exposure produces changes in central monoamines, which are not associated on a dose-dependent basis with major alterations in plasma ACTH.

Effects of sodium arsenite exposure on development and behavior in the rat.

Arsenic is an environmental contaminant found in soil, water and air in some zones of the world. It has been widely studied for its effects as a human carcinogenic agent, but few studies have dealt with neurobehavioral effects. In addition, studies of arsenic effects on development have only addressed its effects on embryotoxicity and teratogenicity after a single oral, gavage or intraperitoneal exposure. Among the behavioral alterations reported after intoxication with arsenic are both increased and decreased locomotor activity and learning deficits in a delayed alternation task [Toxicol. Lett. 54 (1990) 345; Bull. Environ. Contam. Toxicol. 50 (1993) 100; Brain Res. Bull. 55 (2001) 301]. To further characterize developmental and behavioral alterations induced by arsenic exposure, Sprague-Dawley rats were exposed to arsenite (36.70 mg arsenic/l in drinking water) from gestation day 15 (GD 15) or postnatal day 1 (PND 1), until approximately 4 months old. The pregnant or lactating dams received either the arsenic solution or regular drinking water and once pups were weaned, they continued receiving the same solution as drinking water. Animals exposed from GD 15 showed increased spontaneous locomotor activity and both exposed groups showed increased number of errors in a delayed alternation task in comparison to the control group. Total arsenic (TA) content in brain was similar for both exposed groups and significantly different from the control group. These results indicate that rats exposed to arsenic during development present deficits in spontaneous locomotor activity and alterations in a spatial learning task.

Interaction of cadmium with actin microfilaments in vitro.

Cadmium, an environmental and occupational health hazard, can substitute for calcium in the activation of calmodulin in several in vitro assays. It was shown that Cd(2+) can substitute for Ca(2+) in the induction of actin-based gelation in cytoplasmic extracts from rat liver; gelation induced by either Ca(2+) or Cd(2+) is inhibited by trifluoperazine, a well known calmodulin inhibitor; and in MDCK cells there is a Cd(2+)-induced redistribution of actin filaments with the loss of stress fibres and the appearance of actin bundles at the periphery of these cells.

Toxicological assessment of azarcon, a lead salt used as a folk remedy in Mexico. I. Oral toxicity in rats.

Azarcon, a lead tetroxide salt, is used among Mexican and Mexican-American populations for the treatment of digestive illness. Chemical analysis of the azarcon sample used in this study showed it to be 96% lead, 1% calcium, 1% other minor metals, and 2% unidentified material. Taking into account the fact that Pb absorption was estimated at 2% following a single oral administration of 100 mg/kg (Aungst et al., 1981), it is possible to propose a chemical interaction between the components of azarcon, and as a result, the toxicity of Pb tetroxide would be different when given as azarcon than when given as a pure compound. The present work studied this possibility, with the following results. When the treatments of equal doses of pure Pb tetroxide and azarcon were compared (158 mg/kg/day p.o. for 96 h), five of nine tissues studied had similar Pb concentrations. However, with the pure compound the Pb levels were higher in bone and intestines; while with azarcon the Pb levels were higher in heart and brain. The pure Pb tetroxide treatment affects lipid peroxidation only in liver, but a low induction of peroxidation was found also in kidney and heart in rats which received the azarcon treatment. Liver and kidney damage were evident in rats treated with a high dose of azarcon (1.1 g/kg/day p.o. for 96 h), while the effects with the pure compound were similar in type but lower in magnitude. Pb tetroxide as a pure compound inhibits ALA-D by 26% while an inhibition of 42% was found with azarcon.(ABSTRACT TRUNCATED AT 250 WORDS)

Use of the Rey-Osterrieth Complex Figure Test for neurotoxicity evaluation of mixtures in children.

The aim of this study was to assess the value of the children's version of the Rey-Osterrieth Complex Figure Test as a screening test in a population exposed to different mixtures of neurotoxicants. Copy and Immediate Recall scores were evaluated through the test. Children were recruited from three sites; an area with natural contamination by fluoride and arsenic (F-As), a mining-metallurgical area with lead and arsenic contamination (Pb-As) and a malaria zone with the evidence of fish contaminated with dichlorodiphenyltrichloroethane (DDT) and polychlorinated biphenyls (PCBs). Children aged 6-11 years old, living in one of the three polluted sites since birth were recruited (n=166). The exposure was evaluated as follows: fluoride and arsenic in urine, lead in blood and DDT, dichlorodiphenyldichloroethylene (DDE) and PCBs in serum. To evaluate the test performance, z-scores for Copy and Immediate Recall were calculated. The proportion of children by residence area with performance lower than expected by age (below -1 SD) for Copy and Immediate Recall was in the F-As area (88.7% and 59%) and in the DDT-PCBs area (73% and 43.8%), respectively. In the Pb-As area, the proportion was 62% for both tests. After adjustment, Copy correlated inversely with fluoride in urine (r=-0.29; p<0.001) and Immediate Recall correlated inversely with fluoride in urine (r=-0.27; p<0.05), lead in blood (r=-0.72; p<0.01), arsenic in urine (r=-0.63; p<0.05) and DDE (r=-0.25; p<0.05). This study provided evidence that children included in this research are living in high risk areas and were exposed to neurotoxicants. Poor performance in the test could be explained in some way by F, Pb, As or DDE exposure, however social factors or the low quality of school education prevalent in the areas could be playing an important role.

[A method for assessing health risks in mining sites]. / Un método para la evaluación de riesgos para la salud en zonas mineras.

OBJECTIVE: Considering the health risk associated with mining areas, in this work a methodology for the health assessment of this kind of hazardous sites is proposed. MATERIAL AND METHODS: The methodology includes a toxicological assessment, an environmental monitoring of metals, and the exposure assessment of the high risk population. The scheme was evaluated in the mining area of Villa de la Paz, San Luis Potosi, Mexico. The toxicological studies were done in rats treated with mining waste, biomarkers of effect for liver and central nervous tissue were analyzed. Metals levels in surface soil, household dust and water were studied. Finally, urinary arsenic was quantified in children. RESULTS: Neurotoxicity and hepatotoxicity of the mining waste were shown in rats. Then, arsenic and lead levels were analyzed in surface soil, household dust, and water. In all three media, exposure points, heavily contaminated with both metals, were localized. Finally, high levels of urinary arsenic were found in children living in the vicinity of the mine. CONCLUSIONS: Taking into account all these results, the Mexican authorities concluded that a high health risk is present in Villa de la Paz, and a remediation program is in progress.

Arsenic and cadmium exposure in children living near a smelter complex in San Luis Potosí, Mexico.

The main purpose of this study was to assess environmental contamination by arsenic and cadmium in a smelter community (San Luis Potosí City, México) and its possible contribution to an increased body burden of these elements in children. Arsenic and cadmium were found in the environment (air, soil, and household dust, and tap water) as well as in the urine and hair from children. The study was undertaken in three zones: Morales, an urban area close to the smelter complex; Graciano, an urban area 7 km away from the complex; and Mexquitic, a small rural town 25 km away. The environmental study showed that Morales is the most contaminated of the zones studied. The range of arsenic levels in soil (117-1396 ppm), dust (515-2625 ppm), and air (0.13-1.45 micrograms/m3) in the exposed area (Morales) was higher than those in the control areas. Cadmium concentrations were also higher in Morales. Estimates of the arsenic ingestion rate in Morales (1.0-19.8 micrograms/kg/day) were equal to or higher than the reference dose of 1 microgram/kg/day calculated by the Environmental Protection Agency. The range of arsenic levels in urine (69-594 micrograms/g creatinine) and hair (1.4-57.3 micrograms/g) and that of cadmium in hair (0.25-3.5 micrograms/g) indicated that environmental exposure has resulted in an increased body burden of these elements in children, suggesting that children living in Morales are at high risk of suffering adverse health effects if exposure continues.