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Mechanisms of arsenic hyperaccumulation in Pteris vittata. Uptake kinetics, interactions with phosphate, and arsenic speciation.

The mechanisms of arsenic (As) hyperaccumulation in Pteris vittata, the first identified As hyperaccumulator, are unknown. We investigated the interactions of arsenate and phosphate on the uptake and distribution of As and phosphorus (P), and As speciation in P. vittata. In an 18-d hydroponic experiment with varying concentrations of arsenate and phosphate, P. vittata accumulated As in the fronds up to 27,000 mg As kg(-1) dry weight, and the frond As to root As concentration ratio varied between 1.3 and 6.7. Increasing phosphate supply decreased As uptake markedly, with the effect being greater on root As concentration than on shoot As concentration. Increasing arsenate supply decreased the P concentration in the roots, but not in the fronds. Presence of phosphate in the uptake solution decreased arsenate influx markedly, whereas P starvation for 8 d increased the maximum net influx by 2.5-fold. The rate of arsenite uptake was 10% of that for arsenate in the absence of phosphate. Neither P starvation nor the presence of phosphate affected arsenite uptake. Within 8 h, 50% to 78% of the As taken up was distributed to the fronds, with a higher translocation efficiency for arsenite than for arsenate. In fronds, 49% to 94% of the As was extracted with a phosphate buffer (pH 5.6). Speciation analysis using high-performance liquid chromatography-inductively coupled plasma mass spectroscopy showed that >85% of the extracted As was in the form of arsenite, and the remaining mostly as arsenate. We conclude that arsenate is taken up by P. vittata via the phosphate transporters, reduced to arsenite, and sequestered in the fronds primarily as As(III).

Arsenic↗

Docosahexaenoic acid enhances arsenic trioxide-mediated apoptosis in arsenic trioxide-resistant HL-60 cells.

Recent reports indicate a broad spectrum of antileukemic activity for arsenic trioxide (As(2)O(3)) due to its ability to induce apoptosis via intracellular production of reactive oxygen species (ROS). Despite its potent apoptotic mechanism, As(2)O(3) is not equally effective in all leukemic cells, which has prompted a search for agents enhancing As(2)O(3) efficacy. Recently, evidence has been gathered that the polyunsaturated fatty acid docosahexaenoic acid (DHA) may sensitize tumor cells to ROS-inducing anticancer agents. The aim of our investigation was to evaluate whether DHA enhances As(2)O(3)-mediated apoptosis in As(2)O(3)-resistant HL-60 cells. While 1 microM As(2)O(3) or 25 microM DHA reduced cell viability to 85.8% +/- 2.9% and 69.2% +/- 3.6%, combined treatment with As(2)O(3) and DHA reduced viability to 13.0% +/- 9.9% with a concomitant increase of apoptosis. Apoptotic cell death was preceded by collapse of the mitochondrial membrane potential, increased expression of proapoptotic B-cell lymphoma protein-2-associated X protein (Bax), and caspase-3 activation. Importantly, the combined effect of As(2)O(3) and DHA was associated with increased production of intracellular ROS and toxic lipid peroxidation products and was abolished by the antioxidant vitamin E or when oleic acid (a nonperoxidizable fatty acid) was used in place of DHA. Intracellular ROS and toxic lipid peroxidation products most likely constitute the key mediators contributing to the combined effect of As(2)O(3) and DHA. Our data provide the first evidence that DHA may help to extend the therapeutic spectrum of As(2)O(3) and suggest that the combination of As(2)O(3) and DHA could be more broadly applied in leukemia therapy.

Antineoplastic Agents↗

Methylated metabolites of arsenic trioxide are more potent than arsenic trioxide as apoptotic but not differentiation inducers in leukemia and lymphoma cells.

Treatment with arsenic trioxide (As(2)O(3)) by inducing apoptosis and partial differentiation of acute promyelocytic leukemia (APL) cells results in clinical remission in APL patients resistant to chemotherapy and all-trans-retinoic acid. As(2)O(3) (iAs(III)) is methylated in the liver to mono- and dimethylated metabolites, including methylarsonic acid, methylarsonous acid, dimethylarsinic acid, and dimethylarsinous acid. Methylated trivalent metabolites that are potent cytotoxins, genotoxins, and enzyme inhibitors may contribute to the in vivo therapeutic effect of iAs(III). Therefore, we compared the potency of iAs(III) and trivalent metabolites using chemical precursors of methylarsonous acid and dimethylarsinous acid to induce differentiation, growth inhibition, and apoptosis. Methylarsine oxide (MAs(III)O) and to a lesser extent iododimethylarsine were more potent growth inhibitors and apoptotic inducers than iAs(III) in NB4 cells, an APL cell line. This was also observed in K562 human leukemia, lymphoma cell lines, and in primary culture of chronic lymphocytic leukemia cells, but not human bone marrow progenitor cells. Apoptosis was associated with greater hydrogen peroxide accumulation and inhibition of glutathione peroxidase activity. MAs(III)O, in contrast to iAs(III), did not induce PML-retinoic acid receptor alpha degradation, or restore PML nuclear bodies or differentiation in NB4 cells. In a cocultivation experiment, hepatoma-derived HepG2 cells, but not NB4 cells, methylate radiolabeled iAs(III). Methylated metabolites released from HepG2 cells are preferentially accumulated by NB4 cells. This experimental model suggests that in vivo hepatic methylation of iAs(III) may contribute to As(2)O(3)-induced apoptosis but not differentiation of APL cells. MAs(III)O as an apoptotic inducer should be considered in the treatment of other hematologic malignancies like lymphoma.

Apoptosis↗

Toxicity of arsenic species to Lemna gibba L. and the influence of phosphate on arsenic bioavailability.

The toxicity of arsenic (As) species to Lemna gibba L. and the influence of PO(4) (3-) on As bioavailability and uptake were tested in batch culture. L. gibba were exposed to six test concentrations of NaHAsO(4). 7H(2)O and NaAsO(3), with 0, 0.0136, 13.6, and 40 mg L(-1) KH(2)PO(4). In batch culture As toxicity to L. gibba did not relate linearly to As concentration. The growth rate, related to frond number as recommended by OECD and ISO/DIN, was significantly inhibited in fronds exposed to 20-50 microg L(-1) As(III) compared with fronds exposed to As(V). The growth rate was stimulated when plants were exposed to 50-250 microg L(-1) of both As(III) and As(V). After exposure to 300-800 microg L(-1) growth inhibition was significantly higher for As(III) than for As(V), whereas above 800 microg L(-1) As(V) was inhibited the most. The bioaccumulation of As(III) and As(V) was significantly higher for P-deficient cultures (0.98 +/- 0.08 and 1.02 +/- 0.19 g kg(-1), respectively for 0.0136 mg L(-1) PO(4) (3-)) than for P-sufficient cultures (243 and 343 mg kg(-1) for 40 mg L(-1), respectively). Plants exposed to As(V) had uptake and accumulation values slightly higher than did plants exposed to As(III). No significant differences in bioaccumulation were found between plants exposed to a concentration of As(III) >1 mg L(-1) and those exposed to As(V) at the same concentration. This indicates a direct relationship to P content in the culture. Toxicity may result from the uptake of As(V) instead of PO(4) (3-) as a result of ion competition during uptake because of close thermodynamic properties, which may change the interaction among components in the media. The toxicity pattern is interpreted as a manifestation of changing speciation in the batch culture and of the oxidation of As(III) to As(V) in an oxygen-rich environment.

Araceae↗

Nonlinearity in the lung cancer dose-response for airborne arsenic: apparent confounding by year of hire in evaluating lung cancer risks from arsenic exposure in Tacoma smelter workers.

Most analytic studies of human epidemiologic data have affirmed the linear association between excess lung cancer risk and airborne arsenic exposure. Recent Canadian analyses, however, based on the mortality follow-up of Tacoma smelter workers from 1940-1976, provided strong evidence of a nonlinear dose-response when lung cancer risk was expressed in terms of the standardized mortality ratio. Using recently updated data covering ten additional years of mortality experience among Tacoma workers (1940-1986), new analyses were undertaken to further explore nonlinearity in the lung cancer dose-response in this worker cohort. Lung cancer risk was expressed in terms of both the standardized mortality ratio (SMR) and the excess mortality rate (EMR). As in Canadian analyses, nonlinearity was assessed through a three parameter model containing both linear and negative exponential terms for dose. Dropping the negative-exponential dose-term lead to the standard suite of linear dose-response models, with and without intercept, used for comparative purposes. Analyses were undertaken by subcohort as there was strong evidence of confounding by year of initial hire, which largely explained the nonlinearity in the dose-response observed in Canadian analyses. Subcohort analyses based on initial employment, prior to 1940 or thereafter, showed that the nonlinearity in the dose-response was strongly influenced by date of initial hire. whether the cohort risk was measured by either the SMR or EMR, a nonlinear dose-response was evident only among workers hired prior to 1940. This, however, was strongly related to the artifactually low lung cancer mortality seen among workers hired between 1930 and 1939. Among workers hired after 1940, analyses showed that a linear dose-response provided a clearly superior fit. While analyses showed comparable goodness of fit when models were fitted to the SMR and EMR. Only those based on the EMR provided strong evidence of a dose-response. Overall, nonlinearity as observed in Canadian analyses was likely the result of several sources of bias not taken into account by Canadian investigators.

Adult↗

Arsenic content in some Spanish wines. Influence of the wine-making technique on arsenic content in musts and wines.

All of the essential technological means of treating grapes, must and wine were examined systematically. The main reasons for the differences in the arsenic (As) content of rosé and red wines are explained. In this study a relationship has been found between the As level and the wine-making technique. Rosé wines contain more As than red wines because they require a shorter period of contact with the skins. In order to prove this, the average values for the rosé and red wine samples from the same winery were compared.

Arsenic↗

Effect of arsenic on chloroplast ultrastructure and calcium distribution in arsenic hyperaccumulator Pteris vittata L.

This study investigated the impacts of arsenic (As) on the chloroplast ultrastructure and calcium (Ca) distribution in Chinese brake (Pteris vittata L.) mainly by histochemical methods, with an emphasis on the possible function of Ca in As detoxification and accumulation in P. vittata. P. vittata was grown in an artificially contaminated soil added with different concentrations of Na(2)HAsO(4) (0, 100, 300 and 800 mg kg(-1) As dry soil) for 24 weeks in a greenhouse. The addition of As did not affect the chloroplast ultrastructure of young pinna, meanwhile most of the membrane systems of chloroplasts in mature pinna were severely damaged under high As condition. Calcium concentration in the fronds of P. vittata was not significantly affected by the addition of As, but Ca concentration in the mature pinna significantly increased by As addition, consistent with the position appearing As toxicity. When no As was added, most of calcium precipitates distributed around the inner membrane of vacuole. But when the pinna appeared plasmolysis, more calcium precipitates resided outside the cell membrane and bigger particles evenly distributed in the cytoplasm. All the results indicated that Ca had a close relation with As toxicity in P. vittata.

Arsenic↗

Effects of oxalate and phosphate on the release of arsenic from contaminated soils and arsenic accumulation in wheat.

Oxalate is exuded by plants in the rhizosphere and plays an important role in the soil/root interactions. Phosphate fertilizer is widely used all over the world and may influence the behavior of arsenic (As) in soils. In this study oxalate and phosphate were used as extractants to investigate their effects on the release of As from three As-contaminated soils and the chemical speciation of As. Concentrations of arsenite (As(III)) and arsenate (As(V)) released progressively increased by increasing the concentrations of oxalate or phosphate. The released As(V) content was higher than that of As(III) and the differences between As(V) and As(III) released by oxalate was more obvious than by phosphate. Greenhouse experiment was conducted to evaluate the effects of oxalate and phosphate on As uptake by wheat (Triticum vulgare L.). Addition of oxalate or phosphate resulted in the increase of As accumulation in both wheat root and shoot and the effect of phosphate was more obvious than that of oxalate.

Arsenic↗

Arsenic complexes in the arsenic hyperaccumulator Pteris vittata (Chinese brake fern).

Pteris vittata (Chinese brake fern), the first reported arsenic (As) hyperaccumulating plant, can be potentially applied in the phytoremediation As-contaminated sites. Understanding the mechanisms of As tolerance and detoxification in this plant is critical to further enhance its capability of As hyperaccumulation. In this study, an unknown As species, other than arsenite (AsIII) or arsenate (AsV) was found in leaflets by using anion-exchange chromatography-hydride generation-atomic fluorescence spectroscopy and size-exclusion chromatography-atomic fluorescence spectrometry. The chromatographic behavior of this unknown As species and its stability suggest that it is likely an As complex. Although phytochelatin with two subunits (PC2) was the only major thiol in P. vittata under As exposure, this unknown As complex was unlikely to be an AsIII-PC2 complex by comparison of their chromatographic behaviors, stability at different pHs and charge states. The complex is sensitive to temperature and metal ions, but relatively insensitive to pH. In buffer solution of pH 5.9, it is present in a neutral form.

Arsenicals↗

Arsenic hyperaccumulation by Pteris vittata from arsenic contaminated soils and the effect of liming and phosphate fertilisation.

Pot experiments were carried out to investigate the potential of phytoremediation with the arsenic hyperaccumulator Pteris vittata in a range of soils contaminated with As and other heavy metals, and the influence of phosphate and lime additions on As hyperaccumulation by P. vittata. The fern was grown in 5 soils collected from Cornwall (England) containing 67-4550 mg As kg(-1) and different levels of metals. All soils showed a similar distribution pattern of As in different fractions in a sequential extraction, with more than 60% of the total As being associated with the fraction thought to represent amorphous and poorly-crystalline hydrous oxides of Fe and Al. The concentration of As in the fronds ranged from 84 to 3600 mg kg(-1), with 0.9-3.1% of the total soil As being taken up by P. vittata. In one soil which contained 5500 mg Cu kg(-1) and 1242 mg Zn kg(-1), P. vittata suffered from phytotoxicity and accumulated little As (0.002% of total). In a separate experiment, neither phosphate addition (50mg P kg(-1) soil) nor liming (4.6 g CaCO3 kg(-1) soil) was found to affect the As concentration in the fronds of P. vittata, even though phosphate addition increased the As concentration in the soil pore water. Between 4 and 7% of the total soil As was taken up by P. vittata in 4 cuttings in this experiment. The results indicate that P. vittata can hyperaccumulate As from naturally contaminated soils, but may be suitable for phytoremediation only in the moderately contaminated soils.

Arsenic↗

Dynamics of arsenic in agricultural soils irrigated with arsenic contaminated groundwater in Bangladesh.

Arsenic (As) concentrations in the soil layers of 12 rice fields located in four As affected areas and two unaffected areas in Bangladesh were monitored during 2003. In the unaffected areas, where irrigation water contained little As (<1 microg/L), As concentrations of rice field soils ranged from 1.5 to 3.0 mg/kg and did not vary significantly with either depth or sampling time throughout the irrigation period. In the As affected areas where the irrigation water contained elevated As (79 to 436 microg/L), As concentrations of rice field soils were much higher compared to those in the unaffected areas and varied significantly with both depth and sampling time. For the top 0 to 150 mm of the soil, the As concentration increased significantly at the end of the irrigation season (May-June 2003). About 71% of the As that is applied to the rice field with irrigation water accumulates in the top 0 to 75 mm soil layer by the end of the irrigation season. After the wet season during which the rice fields were inundated with flood/rain water, the As concentrations in the soil layer decreased significantly and were reduced to levels comparable to those found in soil samples collected at the beginning of the irrigation period. The long-term As accumulation in agricultural soil appears to be counteracted by biogeochemical pathways leading to As removal from soil.

Agriculture↗

Arsenic-sulfides confound anion exchange resin speciation of aqueous arsenic.

A field-portable anion exchange resin method (often cited as the Ficklin method (1983)) has been extensively used to distinguish between dissolved arsenite (As(III)) and arsenate (As(V)) species in natural waters. As(III) occurs largely as As(OH)(3), which is uncharged at ca. pH 7, while As(V) is negatively charged and will sorb to the resin. However, we show that negatively charged As(III)-sulfide (thioarsenite) species, important at sulfide concentrations >10 microM, also bind to the anion exchange resins, and therefore might be interpreted incorrectly as As(V). Furthermore, we show that nitrogen-purging, which results in a conversion of As(III)-sulfides to arsenite, can be used to obtain accurate arsenic speciation when resins are used on sulfidic water samples.

Adsorption↗

Effects of compost and phosphate amendments on arsenic mobility in soils and arsenic uptake by the hyperaccumulator, Pteris vittata L.

Chinese brake fern (Pteris vittata L.), an arsenic (As) hyperaccumulator, has shown the potential to remediate As-contaminated soils. This study investigated the effects of soil amendments on the leachability of As from soils and As uptake by Chinese brake fern. The ferns were grown for 12 weeks in a chromated-copper-arsenate (CCA) contaminated soil or in As spiked contaminated (ASC) soil. Soils were treated with phosphate rock, municipal solid waste, or biosolid compost. Phosphate amendments significantly enhanced plant As uptake from the two tested soils with frond As concentrations increasing up to 265% relative to the control. After 12 weeks, plants grown in phosphate-amended soil removed >8% of soil As. Replacement of As by P from the soil binding sites was responsible for the enhanced mobility of As and subsequent increased plant uptake. Compost additions facilitated As uptake from the CCA soil, but decreased As uptake from the ASC soil. Elevated As uptake in the compost-treated CCA soil was related to the increase of soil water-soluble As and As(V) transformation into As(III). Reduced As uptake in the ASC soil may be attributed to As adsorption to the compost. Chinese brake fern took up As mainly from the iron-bound fraction in the CCA soil and from the water-soluble/exchangeable As in the ASC soil. Without ferns for As adsorption, compost and phosphate amendments increased As leaching from the CCA soil, but had decreased leaching with ferns when compared to the control. For the ASC soil, treatments reduced As leaching regardless of fern presence. This study suggest that growing Chinese brake fern in conjunction with phosphate amendments increases the effectiveness of remediating As-contaminated soils, by increasing As uptake and decreasing As leaching.

Adsorption↗

A "pennurth of arsenic for rat poison": the Arsenic Act, 1851 and the prevention of secret poisoning.

In this country any chemist or druggist can furnish the means of self-destruction or murder for a few pence, and in too many instances have done so with the utmost indifference. The sale of a poison is regarded as a mere act of commercial intercourse; tant pis for the unfortunate victim of error or passion; he has the benefit of a coroner's inquest; the vendor of the poison receives a reprimand, and things resume their natural course--that is, arsenic and oxalic acid are retailed without compunction, and men are hurried from time to time into eternity.

Animals↗

Bacterial formation of tooeleite and mixed arsenic(III) or arsenic(V)-iron(III) gels in the Carnoulès acid mine drainage, France. A XANES, XRD, and SEM study.

The oxidation of Fe(II) in acid mine drainage (AMD) leads to the precipitation of Fe(III) compounds which may incorporate toxic elements, such as arsenic (As), within their structure or adsorb them at their surface, thus limiting their mobility. The present work provides evidence for spatial and seasonal variations of microbial activity that influence arsenite oxidation and As immobilization in the heavily contaminated AMD from the Carnoulès mine, Gard, France ([As III] = 80 to 280 mg x L(-1) in the acidic spring draining the waste-pile). In the first tens of meters of the AMD, the rapid oxidation of Fe(II) leads to the coprecipitation of large amounts of As with Fe(III) in bacterial mats. XRD, XANES, and SEM analyses of sediments and stromatolite samples revealed the unusual formation of As(III)-rich compounds, especially nanocrystalline tooeleite, Fe6(AsO3)4(SO4)(OH)4 x 4H2O, a rare ferric arsenite sulfate oxy-hydroxide mineral, together with XRD-amorphous mixed As(III)/As(V)-Fe(III) oxy-hydroxide compounds. In the wet season, the suspended sediments of the upstream zone essentially consist of tooeleite associated with am-As(III)-Fe(III) oxy-hydroxides, while am-As(V)-Fe(III) oxy-hydroxides, having As:Fe molar ratios as high as 0.6-0.8, dominate in the dry season. Comparing natural and bioassay samples revealed that the formation of As(III)-rich compounds in the wet season may be related to the metabolic activity of bacterial strains able to oxidize Fe(II) but not As(III). One of these strains, having an Acidithiobacillus ferrooxidans genotype, has been isolated from the Carnoulès AMD. In contrast, the formation of As(V)-rich compounds in the dry season can be related to both biotic and abiotic oxidation of As(III) to As(V). Some Thiomonas strains isolated from the Carnoulès AMD were shown to be able to catalyze the oxidation of As(III) to As(V) in solution. Therefore, they can promote the formation of mixed As(V)-Fe(III) oxy-hydroxides, provided enough Fe(II) oxidizes. These results yield a better understanding of natural processes at this site and may help in designing efficient As-removal processes.

Adsorption↗

Arsenic uptake and accumulation in fern species growing at arsenic-contaminated sites of southern China: field surveys.

Aiming at searching for new arsenic (As) hyperaccumulators, field surveys were conducted at 12 As-contaminated sites located in Guangxi and Guangdong Provinces of southern China. Samples of 24 fern species belonging to 16 genera and 11 families as well as their associated soils were collected and As concentrations in plant and soil samples were determined by inductively coupled plasma-atomic emission spectrometry (ICP-AES). The results show that among 24 fern species, Pteris multifida and P. oshimensis can (hyper)accumulate As in their fronds with high concentrations in addition to P. vittata and P. cretica var. nervosa, which have been previously identified as As hyperaccumulators. Total As concentrations in soils associated with P. multifida and P. oshimensis varied from 1262 to 47,235 mg kg(-1), but the DTPA-extractable As concentrations were relatively low, with a maximum of 65 mg kg(-1). Forty-four of 49 samples of P. multifida collected from five sites and 3 of 13 samples of P. oshimensis collected from one site accumulated over 1000 mg As kg(-1) in their fronds and As concentrations in the fronds were higher than those in the petioles and rhizoids. Although As concentrations in the fronds of P. oshimensis (789 mg kg(-1) averaged, range 301-2142 mg kg(-1)) were comparatively lower than those of P. multifida (1977 mg kg9-1), 624-4056 mg kg(-1)), its high aboveground biomass makes it more suitable for phytoremediating As-contaminated soils. Among all the species in Pteris genus studied, Pteris semipinnata accumulated only very low As concentration in its fronds (8 mg kg(-1), 1-18 mg kg(-1)). Further research is needed to study the differences in As uptake and accumulation among fern species in the same or other genera.

Arsenic↗

Arsenic hyperaccumulation in gametophytes of Pteris vittata. A new model system for analysis of arsenic hyperaccumulation.

The sporophyte of the fern Pteris vittata is known to hyperaccumulate arsenic (As) in its fronds to >1% of its dry weight. Hyperaccumulation of As by plants has been identified as a valuable trait for the development of a practical phytoremediation processes for removal of this potentially toxic trace element from the environment. However, because the sporophyte of P. vittata is a slow growing perennial plant, with a large genome and no developed genetics tools, it is not ideal for investigations into the basic mechanisms underlying As hyperaccumulation in plants. However, like other homosporous ferns, P. vittata produces and releases abundant haploid spores from the parent sporophyte plant which upon germination develop as free-living, autotrophic haploid gametophyte consisting of a small (<1 mm) single-layered sheet of cells. Its small size, rapid growth rate, ease of culture, and haploid genome make the gametophyte a potentially ideal system for the application of both forward and reverse genetics for the study of As hyperaccumulation. Here we report that gametophytes of P. vittata hyperaccumulate As in a similar manner to that previously observed in the sporophyte. Gametophytes are able to grow normally in medium containing 20 mm arsenate and accumulate >2.5% of their dry weight as As. This contrasts with gametophytes of the related nonaccumulating fern Ceratopteris richardii, which die at even low (0.1 mm) As concentrations. Interestingly, gametophytes of the related As accumulator Pityrogramma calomelanos appear to tolerate and accumulate As to intermediate levels compared to P. vittata and C. richardii. Analysis of gametophyte populations from 40 different P. vittata sporophyte plants collected at different sites in Florida also revealed the existence of natural variability in As tolerance but not accumulation. Such observations should open the door to the application of new and powerful genetic tools for the dissection of the molecular mechanisms involved in As hyperaccumulation in P. vittata using gametophytes as an easily manipulated model system.

Arsenates↗

Toenail arsenic concentrations, GSTT1 gene polymorphisms, and arsenic exposure from drinking water.

Toenail arsenic (As) concentrations were evaluated as a biomarker of inorganic As (As(in)) exposure in a population residing in an As-endemic region of Bangladesh. Drinking water and toenail samples were collected from 48 families (n = 223) every 3 months for 2 years and analyzed for As using inductively coupled plasma-mass spectrometry. Drinking water collected 3, 6, and 9 months before each toenail sample collection was combined into a weighted lagged exposure variable. The contribution of each water sample to the measured toenail As concentration was estimated using maximum likelihood that accounted for fluctuations in drinking water exposure and toenail growth. The best model attributed 69%, 14%, and 17% of the toenail As content to drinking water exposures that occurred 3, 6, and 9 months before toenail collection [95% confidence intervals (95% CI), 0.46-0.97, 0.00-0.31, and 0.03-0.35, respectively]. Generalized additive mixed models using penalized regression splines were employed to model the data. Below a drinking water concentration of 2 mug As/L, no relationship between drinking water As and toenail As concentrations was observed. Above this concentration, toenail As content increased in a dose-dependent fashion as drinking water As increased. Age was a significant effect modifier of drinking water As exposure on toenail As (beta = 0.01; 95% CI, 0.002-0.02). Individuals possessing GSTT1-null genotypes had significantly more As in their toenails in contrast to GSTT1 wild-type individuals (beta = 0.11; 95% CI, 0.06-0.2). Therefore, it seems that GSTT1 modifies the relationship between As(in) exposure and toenail As(in) content.

Adolescent↗