Arsenic Binding Research Articles

Dissociation of arsenite‐peptide complexes: Triphasic nature, rate constants, half‐lives, and biological importance

We determined the number and the dissociation rate constants of different complexes formed from arsenite and two peptides containing either one (RVCAVGNDYASGYHYGV for peptide 20) or three cysteines (LECAWQGK CVEGTEHLYSMKCK for peptide 10) via radioactive 73As-labeled arsenite and vacuum filtration methodology. Nonlinear regression analysis of the dissociation of both arsenite-peptide complexes showed that triphasic fits gave excellent r2 values (0.9859 for peptide 20 and 0.9890 for peptide 10). The first phase of arsenite-peptide dissociation had the largest span (decrease in binding), and the rate was too fast to be measured using vacuum filtration methods. The dissociation rate constants of arsenite-peptide complexes for the second phase were 0.35 and 0.54 min(-1) and for the third phase were 0.0071 and 0.0045 min(-1) for peptides 20 and 10, respectively. For peptide 20, the three spans of triphasic decay were 85%, 9%, and 7% of the total binding of 16.1 nmol/mg protein. For peptide 10, which can bind in both an intermolecular and intramolecular manner, the three spans of triphasic decay were 59%, 16%, and 25% of the total binding of 43.7 nmol/mg protein. Binding of trivalent arsenicals to peptides and proteins can alter their structure and function and contribute to adverse health outcomes such as toxicity and carcinogenicity.

Remediation of soil contaminated with arsenic, zinc, and nickel by pilot‐scale soil washing

AbstractThe use of soil washing has successfully demonstrated bench‐scale and field applications for the remediation of oxyanions and cationic heavy metals. However, it is not easy to simultaneously remove both oxyanions and cations using one wash solution in the same extraction. Also contaminated soils from metal mining areas present other difficulties to remove contaminants because of their strong binding properties. Correspondingly, the efficiency of soil washing can be inhibited by the limited extraction of contaminants. The objective of this study was to find the optimal process conditions for the enhanced washing of recalcitrant contaminants. This study focused on the effectiveness of engineering‐scale washing of field soil using three types of inorganic acids (HCl, H2SO4, and H3PO4). In this study, soil contaminated with arsenic, nickel, and zinc was obtained from a closed iron/serpentine mining area. The H2SO4 and H3PO4 wash solutions, including the competitive oxyanion, enhanced the removal of arsenic. The acid attack on arsenic binding in the Fe/Mn oxide and organic/sulfides fractions may be a possible removal mechanism in addition to anionic competitive desorption. The low efficiency of soil washing results from the strong binding associated with minerals and the enrichment of contaminants in the highly reactive fine particles. For this soil used, the physical separation of particles could enhance the overall effectiveness of soil washing. Therefore, for the soils contaminated with recalcitrant contaminants in metal mining areas, the physical separation of fine particles can be significant in addition to the chemical extraction. © 2005 American Institute of Chemical Engineers Environ Prog, 2005

Arsenite binding to synthetic peptides based on the Zn finger region and the estrogen binding region of the human estrogen receptor-α

We selected the estrogen receptor protein for study because of prior results indicating that arsenite is a “potential nonsteroidal environmental estrogen”. We utilized radioactive 73As-labeled arsenite and vacuum filtration methodology to determine the binding affinity of arsenite to synthetic peptides. A zinc finger region containing four free sulfhydryls and the hormone binding region containing three free sulfhydryls based on the human estrogen receptor-α were studied. Peptide 15 (RYCAVCNDYASGYHYGVWSCEGCKA) bound arsenite with a K d of 2.2 μM and B max (maximal binding capacity) of 89 nmol/mg protein. Peptide 10 (LECAWQGKCVEGTEHLYSMKCKNV) had a K d of 1.3 μM and B max of 59 nmol/mg protein. In contrast, the monothiol peptide 19 (LEGAWQGKGVEGTEHLYSMKCKNV) bound arsenite with a higher K d of 124 μM and a B max of 26 nmol/mg protein. In our studies, amino acids other than cysteine (including methionine and histidine) did not bind arsenite at all. Peptides modeled on the estrogen receptor with two or more nearby free sulfhydryls (two or five intervening amino acids) had low K d values in the 1–4 μM range. Peptides containing single sulfhydryls or two sulfhydryls spaced 17 amino acids apart had higher K d values in the 100–200 μM range, demonstrating lower affinity. With the exception of peptide 24 which had an unusually high B max value of 234 nmol/mg, the binding capacity of the studied peptides was proportional to the number of free cysteines. Binding of trivalent arsenicals to peptides and proteins can contribute to arsenic toxicity and carcinogenicity via altered peptide/protein structure and enzyme function.

Use of hydride generation-atomic absorption spectrometry to determine the effects of hard ions, iron salts and humic substances on arsenic sorption to sorghum biomass

At present, there is a great interest in studying new sorbent materials for the removal of arsenic from aqueous solutions because of its high toxicity and adverse effects on human health. In previous research, sorghum biomass was found to be an efficient and economic sorbent for the removal of arsenic from aqueous solutions. In this investigation, the effects of CaCl 2, MgCl 2, FeSO 4, MgSO 4, Fe(NO 3) 3, and humic substances (peat moss, humin and humic acids) on arsenic binding to sorghum biomass were evaluated. Among these compounds, only iron salts were found to positively increase the sorption of arsenic to sorghum biomass. In addition, the sorption equilibrium was reached faster when the reaction mixture contained iron salts. However, an overall reduction of 21% of arsenic sorption to sorghum biomass was observed in the presence of MgSO 4. This interference may be due to the presence of sulfate ions, instead of the hard cations, that could be in competition with As for the same interaction sites or ligands. Peat moss, humins and humic acid, extracted from sphagnum peat moss, significantly decreased the arsenic sorption to sorghum biomass.

Fractionation of sedimentary arsenic from Port Kembla Harbour, NSW, Australia

The binding of arsenic in sediments of the heavily industrialised Port Kembla Harbour, NSW, Australia, has been investigated. Both dredge and core samples have been used to develop a sieving/sequential extraction (SE) procedure. Dredge samples included oxic surficial and deeper anoxic sediment. The main core sample analysed was 18 cm deep, sliced at 2 cm intervals. Sediment was sieved to three size ranges (<63 microm, 63-250 microm, >250 microm) and each of these was then subjected to a four step SE, sequentially solubilizing arsenic as ion exchangeable, 1 M HCl soluble, NH(2)OH.HCl soluble, and strong oxidising acid soluble. Concentrations of 50-500 mg As kg(-1) were found, elevated well above local background values. The core sample showed elevated concentrations of arsenic within the top 6-8 cm (300-500 mg As kg(-1)), relative to the deeper sediment (100-200 mg As kg(-1)). Substantial portions of the total arsenic present in the 0-8 cm sediments of core and dredge samples, were found to be soluble in 1 M pH 5 phosphate buffer or 1 M HCl. Arsenic in the lower 8-18 cm of the core displayed different solubility, the fourth stage SE strong acid digestion being required to solubilize >90% of the deep sediment arsenic. It appears that diagenesis had resulted in remobilisation of weakly bound arsenic with subsequent diffusion and deposition in surficial layers. Strong acid soluble arsenic present in deeper sediments has two possible origins: sedimented as strongly bound remaining untouched by diagenetic events, or subjected to diagenetic reactions such as pyritization, which lead to more stable crystalline forms of minerals.

Distribution of elements binding to molecules with different molecular weights in aqueous extract of Antarctic krill by size-exclusion chromatography coupled with inductively coupled plasma mass spectrometry

The distribution of silver, arsenic, cadmium, cobalt, chromium, copper, iron, manganese, nickel, lead, selenium and zinc binding to species with different molecular weight in aqueous extract of krill was studied by on-line size-exclusion chromatography (SEC)/inductively coupled plasma mass spectrometry (ICP-MS). The extract was fractionated in three fractions with different molecular weight (MW) ranges (>20,000 relative molecular mass (rel. mol. mass), 2000–20,000 rel. mol. mass and <2000 rel. mol. mass), which were further analyzed by SEC with columns having different optimum fractionation ranges in order to obtain more detailed information about the MW distribution of the elements. Various distribution profiles for the target elements among different MW ranges were observed. The results obtained indicated that manganese, zinc, silver, cadmium and lead species were mostly distributed in the higher MW range (>20,000 rel. mol. mass). In the case of chromium, iron, cobalt, arsenic and selenium, most of them bind to species with lower MW (<2000 rel. mol. mass). Only copper and nickel species was predominantly present in middle MW range (2000–20,000 rel. mol. mass). Further speciation of arsenic compounds in the small MW fraction was carried out with anion exchange chromatography (AEC) coupled with ICP-MS. The results showed that the dominant arsenic species in this fraction is As(III) (63% of extractable arsenic), while As(V) (13%) and two unknown arsenic species (19% and 5%, respectively) are present in lower amounts.

Arsenic binding to Fucus vesiculosus metallothionein

The seaweed Fucus vesiculosus is a member of the brown algae family. Kille and co-workers [Biochem. J. 338 (1999) 553] reported that this species contains the gene for metallothionein. Metallothionein is a metalloprotein having low molecular weight, and high cysteine content, which binds a range of metals. F. vesiculosus bioaccumulates arsenic from the aquatic environment [Mar. Chem. 18 (1986) 321]. In this paper we describe arsenic binding to F. vesiculosus metallothionein, characterized by electrospray ionization mass spectrometry. Five arsenic-MT species were detected with increasing As to protein ratios. These results provide important information about the metal-chelation behaviour of this novel algal metallothionein which is a putative model for arsenic binding to F. vesiculosus in vivo.

Equilibrium characterization of the As(III)?cysteine and the As(III)?glutathione systems in aqueous solution

Some arsenic compounds were the first antimicrobial agents specifically synthesized for the treatment of infectious diseases such as syphilis and trypanosomiasis. More recently, arsenic trioxide has been shown to be efficient in the treatment of acute promyelocytic leukemia. The exact mechanism of action has not been elucidated yet, but it seems to be related to arsenic binding to vicinal thiol groups of regulatory proteins. Glutathione is the major intracellular thiol and plays important roles in the cellular defense and metabolism. This paper reports on a study of the interactions between arsenic(III) and either cysteine or glutathione in aqueous solution. The behavior observed for the As(III)–glutathione system is very similar to that of As(III)–cysteine. In both cases, the formation of two complexes in aqueous solution was evidenced by NMR and electronic spectroscopies and by potentiometry. The formation constants of the cysteine complexes [As(H−1Cys)3], logK=29.84(6), and [As(H−2Cys)(OH)2]−, logK=12.01(9), and of the glutathione complexes [As(H−2GS)3]3−, logK=32.0(6), and [As(H−3GS)(OH)2]2−, logK=10(3) were calculated from potentiometric and spectroscopic data. In both cases, the [As(HL)3] species, in which the amine groups are protonated, predominate from acidic to neutral media, and the [As(L)(OH)2] species appear in basic medium (the charges were omitted for the sake of simplicity). Spectroscopic data clearly show that the arsenite-binding site in both complexes is the sulfur atom of cysteine. In the [As(L)(OH)2] species, the coordination sphere is completed by two hydroxyl groups. In both cases, arsenic probably adopts a trigonal pyramidal geometry. Above pH 10, the formation of [As(OH)2O]− excludes the thiolates from arsenic coordination sites. At physiological pH, almost 80% of the ligand is present as [As(HL)3].

Equilibrium characterization of the As(III)–cysteine and the As(III)–glutathione systems in aqueous solution

Some arsenic compounds were the first antimicrobial agents specifically synthesized for the treatment of infectious diseases such as syphilis and trypanosomiasis. More recently, arsenic trioxide has been shown to be efficient in the treatment of acute promyelocytic leukemia. The exact mechanism of action has not been elucidated yet, but it seems to be related to arsenic binding to vicinal thiol groups of regulatory proteins. Glutathione is the major intracellular thiol and plays important roles in the cellular defense and metabolism. This paper reports on a study of the interactions between arsenic(III) and either cysteine or glutathione in aqueous solution. The behavior observed for the As(III)–glutathione system is very similar to that of As(III)–cysteine. In both cases, the formation of two complexes in aqueous solution was evidenced by NMR and electronic spectroscopies and by potentiometry. The formation constants of the cysteine complexes [As(H −1Cys) 3], log K=29.84(6), and [As(H −2Cys)(OH) 2] −, log K=12.01(9), and of the glutathione complexes [As(H −2GS) 3] 3−, log K=32.0(6), and [As(H −3GS)(OH) 2] 2−, log K=10(3) were calculated from potentiometric and spectroscopic data. In both cases, the [As(HL) 3] species, in which the amine groups are protonated, predominate from acidic to neutral media, and the [As(L)(OH) 2] species appear in basic medium (the charges were omitted for the sake of simplicity). Spectroscopic data clearly show that the arsenite-binding site in both complexes is the sulfur atom of cysteine. In the [As(L)(OH) 2] species, the coordination sphere is completed by two hydroxyl groups. In both cases, arsenic probably adopts a trigonal pyramidal geometry. Above pH 10, the formation of [As(OH) 2O] − excludes the thiolates from arsenic coordination sites. At physiological pH, almost 80% of the ligand is present as [As(HL) 3].

Effects of As(III) Binding on α-Helical Structure

As(III) displays a wide range of effects in cellular chemistry. Surprisingly, the structural consequences of arsenic binding to peptides and proteins are poorly understood. This study utilizes model alpha-helical peptides containing two cysteine (Cys) residues in various sequential arrangements and spatial locations to study the structural effects of arsenic binding. With i, and i + 1, i + 2, or i + 3 arrangements, CD spectroscopy shows that As(III) coordination causes helical destabilization when Cys residues are located at central or C-terminal regions of the helix. Interestingly, arsenic binding to i, i + 3 positions results in the elimination of helical structure and the formation of a relatively stable alternate fold. In contrast, helical stabilization is observed for peptides containing i, i + 4 Cys residues, with corresponding pseudo pairwise interaction energies (Delta G(pw) degrees) of -1.0 and -0.7 kcal/mol for C-terminal and central placements, respectively. Binding affinities and association rate constants show that As(III) binding is comparatively insensitive to the location of the Cys residues within these moderately stable helices. These data demonstrate that As(III) binding can be a significant modulator of helical secondary structure.

Comparison of the binding of cadmium(II), mercury(II), and arsenic(III) to the de novo designed peptides TRI L12C and TRI L16C.

Designed alpha-helical peptides of the TRI family with a general sequence Ac-G(LKALEEK)(4)G-CONH(2) were used as model systems for the study of metal-protein interactions. Variants containing cysteine residues in positions 12 (TRI L12C) and 16 (TRI L16C) were used for the metal binding studies. Cd(II) binding was investigated, and the results were compared with previous and current work on Hg(II) and As(III) binding. The metal peptide assemblies were studied with the use of UV, CD, EXAFS, (113)Cd NMR, and (111m)Cd perturbed angular correlation spectroscopy. The metalated peptide aggregates exhibited pH-dependent behavior. At high pH values, Cd(II) was bound to the three sulfurs of the three-stranded alpha-helical coiled coils. A mixture of two species was observed, including Cd(II) in a trigonal planar geometry. The complexes have UV bands at 231 nm (20 600 M(-1) cm(-1)) for TRI L12C and 232 nm (22 600 M(-1) cm(-1)) for TRI L16C, an average Cd-S bond length of 2.49 A for both cases, and a (113)Cd NMR chemical shift at 619 ppm (Cd(II)(TRI L12C)(3)(-)) or 625 ppm (Cd(II)(TRI-L16C)(3)(-)). Nuclear quadrupole interactions show that two different Cd species are present for both peptides. One species with omega(0) = 0.45 rad/ns and low eta is attributed to a trigonal planar Cd-(Cys)(3) site. The other, with a smaller omega(0), is attributed to a four-coordinate Cd(Cys)(3)(H(2)O) species. At low pH, no metal binding was observed. Hg(II) binding to TRI L12C was also found to be pH dependent, and a 3:1 sulfur-to-mercury(II) species was observed at pH 9.4. These metal peptide complexes provide insight into heavy metal binding and metalloregulatory proteins such as MerR or CadC.

Male reproductive effect of arsenic in mice.

Arsenic, a known human carcinogen, was given to mice via drinking water as sodium arsenite at a dose 53.39, 133.47, 266.95 and 533.90 micromol 1 for 35 days. A decrease in the activity of 17 beta HSD along with increase in LDH, gammaGT activity were observed at 533.90 micromol l. The observed sperm count, motility and morphological abnormalities in sperm were similar to control at lower dose levels. However at 533.90 micromol l a significant decrease in sperm count and motility along with increase in abnormal sperm were noticed. Significant accumulation of arsenic in testes and accessory sex organs may be attributed to the arsenic binding to the tissues or greater cellular uptake. No effects were observed on indices studied for reproductive effects at 53.39 micromol l arsenic close to which human being are exposed through drinking water under the present set of experimental conditions.

Arsenic binding proteins from human lymphoblastoid cells

Arsenic is a ubiquitous contaminant of drinking water and food. The mechanisms of the toxic action of inorganic arsenic are unknown. We report the isolation of proteins having a high affinity for arsenic in the +3 oxidation state that are induced by arsenite (AsIII) in human lymphoblastoid cells. The arsenic-binding proteins were isolated using a p-aminophenylarsine oxide affinity column. At least four proteins of 50, 42, 38.5 and 19.5 kDa were isolated by elution with 10 or 100 mM 2-mercaptoethanol. Two proteins were tentatively identified as tubulin and actin on the basis of their molecular weights and previously reported affinity for the arsenic column. The identities of the remaining proteins are unknown. Heme oxygenase 1 was induced by AsIII but did not bind to the arsenic affinity column. We conclude that AsIII induces multiple proteins that have variable affinities for arsenic in the +3 state as judged by the concentration of 2-mercaptoethanol required for their elution. The arsenic binding motif of these proteins may involve three thiol groups arranged 3–6 Å apart by the tertiary structure of the protein as suggested by others. These proteins may serve as high affinity binding sites for AsIII and may be involved in the biological action of AsIII.

Binding of Arsenicals to Proteins in anin VitroMethylation System

The dynamics of interactions between rat liver cytosolic proteins and arsenicals were examined in anin vitromethylation system that contained cytosol, glutathione,S-adenosylmethionine, and 1 μm[73As]arsenite. After incubation at 37°C for up to 90 min, low-molecular-weight components of the assay system (<10 kDa) were removed by ultrafiltration and cytosolic proteins were separated by size-exclusion chromatography on Sephacryl S-300 gel. Five73As-labeled protein peaks were found in chromatographic profiles. The estimated molecular masses of73As-labeled proteins eluting in the three earliest peaks were as follows:Vo, ≥1000 kDa; A, 135 kDa; and B, 38 kDa. Peak C eluted immediately before the total volume (VT) of the chromatographic column; peak D eluted after theVT.73As bound to proteins was released by CuCl treatment and speciated by thin-layer chromatography. Amounts and ratios of inorganic As, methyl As, and dimethyl As associated with cytosolic proteins depended upon the incubation interval. Inorganic As was present in all protein peaks. Methyl As was primarily associated with peaks A and C; dimethyl As was associated with peaks B and C. To examine the effect of valence on the binding of methylarsenicals to cytosolic proteins, trivalent or pentavalent14C-labeled methyl As or dimethyl As was incubated in anin vitrosystem designed to minimize the enzymatically catalyzed production of methylated arsenicals. Proteins in peaks A, B, and C bound preferentially trivalent methyl and dimethyl As. Peak D bound either trivalent or pentavalent methyl and dimethyl As. Protein-bound inorganic and methyl As were substrates for the production of dimethyl As in anin vitromethylation system, suggesting a role for protein-bound arsenicals in the biomethylation of this metalloid.

Binding of phenylarsenoxide to Arg-tRNA protein transferase is independent of vicinal thiols.

Reversible enzyme inhibition by phenylarsenoxides is generally taken to indicate the presence of functionally important vicinal thiol groups. Arginyl aminoacyl-tRNA transferase from eukaryotes is potently inhibited by phenylarsenoxides and possesses one or more essential sulfhydryl groups (Li, J., & Pickart C. M. (1995) Biochemistry 34, 139-147]. To map the putative Cys residues that mediate arsenoxide binding to the transferase from Saccharomyces cerevisiae, we systematically mutagenized the 15 Cys residues of the transferase, singly and in combination, to Ala (13 Cys) or Ser (2 Cys). Six mutant enzymes, encompassing all 15 Cys residues of the transferase, were characterized in detail. The results revealed that Cys-20, Cys-23, and Cys-94 and/or Cys-95 were important for activity, since mutations at these positions reduced activity by 100-fold (Cys-94 and Cys-95 were mutated simultaneously). Surprisingly, however, all of the mutant enzymes retained the ability to bind a radioiodinated phenylarsenoxide derivative, with undiminished stoichiometry and affinity. All of the mutant enzymes also remained susceptible to irreversible reaction with a bifunctional phenylarsenoxide bearing a paraalkyl halide substituent. Prior reaction of the enzyme with the bifunctional reagent blocked subsequent binding of the radiolabeled phenylarsenoxide, indicating that these two reagents bind at a single common site. These results indicate that high-affinity binding of trivalent arsenicals can occur by a thiol-independent mechanism.

Interactions of rat red blood cell sulfhydryls with arsenate and arsenite.

Arsenic-thiol interactions were investigated by determining changes in rat blood sulfhydryls after exposure to arsenate, As(V), or arsenite, As(III). Incubation with As(V) resulted in time- and dose-dependent depletion of nonprotein sulfhydryls (NPSH), specifically glutathione (GSH). At the highest As(V) concentration (10 mM), significant loss of glutathione was only observed after 3 h of incubation, but by 5 h 0.5 mM As(V) and higher was sufficient to deplete GSH. As(V) was reduced to As(III) at all dose levels, indicating a redox interaction with GSH, but oxidized glutathione (GSSG) was not formed in sufficient quantities to account for losses in GSH. This may be due to formation of another oxidized species such as a protein-mixed-disulfide (ProSSG). Further evidence that glutathione reduces arsenate was obtained by pretreating cells with the sulfhydryl derivatizing agent N-ethylmaleimide (NEM). Removal of thiols with NEM severely inhibited the formation of As(III) in these incubations, indicating that the main pathway for arsenate reduction in red cells is sulfhydryl dependent. As(III) demonstrated a completely different profile of sulfhydryl interaction. Sulfhydryls (NPSH and GSH) were depleted but the losses were primarily accounted for by oxidation to GSSG. As(III) was also a more potent sulfhydryl depleting agent, requiring only 0.1 mM As(III) to significantly reduce GSH after 5 h of incubation. Significant levels of GSSG formed at all doses of As(III). Evidence is presented to suggest that As(III) also formed mixed complexes with protein and glutathione. Samples that were acid precipitated displayed loss of cytosolic glutathione, which could be reversed if NEM was added prior to protein precipitation. Arsenic was detected in high quantities in the protein precipitates, and this was also found to be reversible by NEM treatment. The fact that both GSH depletion and protein binding were reversible by NEM treatment points to formation of a mixed complex of protein, GSH, and As(III), possibly ProS-As-(SG)x. Arsenic affinity chromatography and polyacrylamide gel electrophoresis were used to characterize arsenic binding proteins in red-cell cytosol. The main arsenic binding protein appeared to be hemoglobin.

Arsenic binding proteins of mammalian systems: I. Isolation of three arsenite-binding proteins of rabbit liver

It is well known that arsenite/arsenate (As 3+/As 5+) administered to rabbits is bound initially to cellular proteins of the liver before methylated arsenic metabolites appear in urine. This protein binding may decrease the in situ toxicity of inorganic arsenic by decreasing its metabolic availability until it is methylated enzymatically. We have investigated the binding of As 3+ and As 5+ to the cytosolic proteins of rabbit liver. The results indicate that when cytosolic proteins are incubated with inorganic arsenic, the amount of As 3+ bound is 13 times greater than that for As 5+. Arsenite-specific binding sites on cytosolic proteins were determined to be 67% of the total (specific and non-specific) number of possible binding sites. Ammonium sulfate fractionation, non-denaturing PAGE and gel filtration chromatography indicate that three liver proteins with molecular weights of 100 kDa, 450 kDa and > 2000 kDa strongly bind arsenite. The radioactive profiles after gel filtration chromatography of liver cytosolic proteins are very similar whether As 3+ binding occurs in vitro or in vivo. Thus, the in vitro model appears to be valid for further study of these arsenite-binding proteins.

Interaction of selenium and arsenic with metallothionein: effect of vitamin B12.

The effects of selenium and arsenic on metallothionein (MT) levels were determined in vitamin B12-supplemented and depleted rats. The binding of selenium and arsenic to MT in vitro and in vivo was also investigated. Rats fed a vitamin B12-deficient diet had significant (p < 0.05), higher levels of liver MT as compared to vitamin B12-supplemented rats. Rats fed 5-9 micrograms/g selenium as selenite or 50-150 micrograms/g arsenic as arsenite showed no significant increase of MT content in the liver. However, significant increases in MT levels in liver (p < 0.01) and kidney (p < 0.05) were seen when rats were injected with 1.5 mg selenium as selenite, and increase of liver MT levels (p < 0.05) was shown when injected with 5 mg arsenic as arsenite/kg body weight. In comparison to zinc, injection of selenium or arsenic caused only a slight elevation of liver MT levels. Liver cytosolic Sephadex G-75 patterns of rats injected with both zinc and arsenic gave separate peaks for arsenic and zinc which suggest that in vivo arsenic binding to MT may be insignificant. Spectrophotometric absorption changes at 255 nm with various levels of selenium or arsenic additions to an MT solution were used to show that selenite gave a maximum absorption increase at a 1:1 molar ratio, but maximum absorption change occurred with arsenite at a molar ratio of 7:1.

In Vivo and in Vitro Percutaneous Absorption and Skin Decontamination of Arsenic from Water and Soil

The objective was to determine the percutaneous absorption of arsenic-73 as H3AsO4 from water and soil. Soil (Yolo County 65-California-57-8) was passed through 10-, 20-, and 48-mesh sieves. Soil retained by 80 mesh was mixed with radioactive arsenic-73 at a low (trace) level of 0.0004 μg/cm2 (micrograms arsenic per square centimeter skin surface area) and a higher dose of 0.6 μg/cm2. Water solutions of arsenic-73 at a low (trace) level of 0.000024 μg/cm2 and a higher dose of 2.1 μg/cm2 were prepared for comparative analysis. In vivo in Rhesus monkey a total of 80.1 ± 6.7% (SD) intravenous arsenic-73 dose was recovered in urine over 7 days; the majority of the dose was excreted in the first day. With topical administration for 24 hr, absorption of the low dose from water was 6.4 ± 3.9% and 2.0 ± 1.2% from the high dose. In vitro percutaneous absorption of the low dose from water with human skin resulted in 24-hr receptor fluid (phosphate-buffered saline) accumulation of 0.93 ± 1.1% dose and skin concentration (after washing) of 0.98 ± 0.96%. Combining receptor fluid accumulation and skin concentration gave a combined amount of 1.9%, a value less than that in vivo (6.4%) in the Rhesus monkey. From soil, receptor fluid accumulation was 0.43 ± 0.54% and skin concentration was 0.33 ± 0.25%. Combining receptor fluid plus skin concentrations gave an absorption value of 0.8%, an amount less than that with in vivo absorption (4.5%) in the Rhesus. These absorption values did not match current EPA default assumptions. Washing with soap and water readily removed residual skin surface arsenic, both in vitro and in vivo. The partition coefficient of arsenic in water to powdered human stratum corneum was 1.1 × 104and from water to soil it was 2.5 × 104. This relative similarity in arsenic binding to powdered human stratum corneum and soil may indicate why arsenic absorption was similar from water and soil. This powdered human stratum corneum partition coefficient model may provide a facile method for such predictions.

The role of the methylation in the detoxication of arsenate in the rabbit

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.