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Projected uptake and toxicity of selenium compounds from the environment.

Industrial workers and members of the general public may be exposed to selenium by inhalation of selenium in the workplace or atmosphere or by ingestion of selenium in food. A model has been developed to evaluate the potential uptake of selenium in body tissues by these two exposure routes. Rates were estimated for transport of selenium between five compartments including lung, gastrointestinal tract, blood, liver and other tissues. Results of model simulations were compared to published tissue distribution information obtained from single inhalation exposures of rats and dogs to radiolabeled selenium compounds at concentrations from 20 mg/m3 to 20 micrograms/m3 with initial body burdens of selenium ranging from 28 to 0.09 micrograms Se/kg body wt. The model was then modified to predict equilibrium organ concentrations of selenium in people after continual exposure to selenium in the air or in the diet. Daily intake levels of 100 micrograms/day and a fractional absorption value of 0.8 were used. With an air concentration of 1 ng Se/m3, model predictions indicated that most of the total body selenium in people is likely to come from their diet because selenium in the urban atmosphere contributes a very small part of the total body selenium. However, continual inhalation of selenium at the threshold limit value (TLV; 200 micrograms/m3) could contribute significantly to the total body burden of selenium. Levels of selenium predicted in lung, liver, and blood after inhalation of selenium at the TLV were 22,000, 1200, and 440 ng Se/g tissue. Predicted lung concentrations were near those that produced toxic effects in animals after ingestion of Se.

Absorption↗

Effects of selenium compounds on formate metabolism and coincidence of selenium-75 incorporation and formic dehydrogenase activity in cell-free preparations of Escherichia coli.

The effects of selenite, selenocystine, and selenomethionine in a defined growth medium on formic dehydrogenase biosynthesis in aerobically and anaerobically grown Escherichia coli have been studied. Sucrose gradient centrifugation of a partially purified enzyme demonstrated a coincidence of (75)Se incorporation and formic dehydrogenase activity.

Aerobiosis↗

Antitumor effect of a new organic selenium compound, 6-phenyl-7 (6H)-isoselenazolo [4,3-d] pyrimidone (ISP), on the growth of P388 mouse leukemia.

The organic selenium compound, ISP [6-phenyl-7(6H)-isoselenazolo [4,3-d] pyrimidone] markedly inhibited the growth of P388 mouse leukemia at dose of 100 micrograms/mouse per day x 10 with no sign of toxicity. The antitumor activity of 4,5-dihydro-4-methyl-6-oxo-5-phenyl-6H-pyrazolo [4,5-c] isoselenazole (PIS) was weaker than that of ISP. The total lipid and phospholipid contents in the leukemic cells treated with ISP were significantly decreased. The fatty acid pattern of cholesterol esters, phosphatidyl choline and phosphatidyl ethanolamine from the ISP-treated P388 leukemic cells differed markedly from that of the corresponding lipids from the control leukemic cells. In addition, the synthesis of DNA or RNA was depressed in the ISP-treated leukemic cells. The present results indicate that ISP may open new perspectives in cancer chemotherapy.

Amides↗

Fractionation of soluble selenium compounds from fish using size-exclusion chromatography with on-line detection by inductively coupled plasma mass spectrometry.

Fish accumulate significant amounts of selenium and are an important dietary source of this element. Some studies have however indicated a low bioavailability of the selenium from fish. Since little is known of the selenium forms in fish, we have studied soluble selenium compounds in fish species, and compared different techniques for fractionation of selenocompounds (size-exclusion chromatography, ultrafiltration, and precipitation with trichloroacetic acid). The size-exclusion column (Superdex 200 HR 10/30) was coupled on-line to inductively coupled plasma mass spectrometry (ICP-MS). The limit of detection was 0.20 microgram l-1 and the selenium response was linear in the investigated concentration range of 0-20 micrograms l-1 (r2 = 0.98). For plaice 47% of the selenium was extractable while the extraction efficiency for cod was 23%. The fish extracts were injected onto the column four times each and the variation in the quantitative data for different selenium-containing fractions between the runs was small (RSD < 10%). The recovery of selenium in the chromatographic step was about 70%, indicating some interaction between the fish extracts and the column material. Ultrafiltration using a membrane with a cut-off at M(r) 10,000 gave results similar to the size-exclusion fractionation, for cod about 20% of the soluble selenium had a M(r) < 10,000 and the corresponding value for plaice was 69%. Removal of high-molecular-weight compounds from the sample by trichloroacetic acid precipitation showed a similar proportion of low-molecular-weight compounds for plaice (77%), while the obtained value for cod was higher (38%) compared with the other techniques.

Animals↗

Effect of selenium compounds on murine B16 melanoma cells and pigmented cloned pB16 cells.

The effects of selenium compounds such as sodium selenite, sodium selenate, seleno-DL-cystine and seleno-DL-methionine (100 microM and 10 microM) on B16 and pigmented cloned pB16 murine melanoma cells were investigated in vitro. At the tested concentrations, B16 cells showed a greater sensitivity to the toxic effects of sodium selenite and seleno-DL-cystine than pB16 cells, whereas no decrease of B16 and pB16 cell number was observed after incubation with sodium selenate or seleno-DL-methionine. Glutathione (GSH) percentages were strongly decreased only by selenite and seleno-DL-cystine; it was marked more in B16 than in pB16 cells. The pretreatment of B16 cells with a GSH depleting agent (10 microM buthionine-[S,R]-sulfoximine) did not significantly influence the cytotoxic effects of selenite and seleno-DL-cystine. On both cell populations, GSH preincubation (50 microM) enhanced the cytotoxicity of selenite whereas the survival of seleno-DL-cystine treated cells was increased. Glutathione peroxidase (GSH-Px) activity in B16 cells was more sensitive than in pB16 cells to the activating effect of selenite, and particularly of seleno-DL-cystine: however, cell-free controls indicated that activation was mainly due to glutathione reductase. The rate of 75Se (as sodium selenite) uptake in both cell populations was maximal within the first hour of incubation, with a preferential accumulation in the cytosol; after 24 h of incubation, the amount of 75Se in cytosol and pellet was approximately the same.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Retrograde tracing of zinc-containing neurons by selenide ions: a survey of seven selenium compounds.

The autometallographic retrograde tracing of zinc-containing neurons by intracerebral injection of sodium selenite (Na2SeO3), introduced by Danscher in 1982, has recently been described in more detail. Intracerebral injections of both sodium selenide (Na2Se) and sodium selenite (Na2SeO3) have been successfully used; however, sodium selenite had a rather toxic effect on the injected tissue. In the present study, we tested seven different selenium compounds to find the most suitable compound for retrograde tracing of zinc-positive pathways. Among the tested compounds, sodium selenide (Na2Se) caused insignificant necrosis within the injection site and was easily transported retrogradely when handled anaerobically. Sodium selenide is therefore recommended as the compound of choice.

Animals↗

Metabolic differences and similarities of selenium in blood and brain of the rat following the administration of different selenium compounds.

A common intermediate, i.e., selenite, was found in the serum of the rat; the maximum levels occurred 3 h after administration independent of chemical forms. This indicates that both the reduction of selenate to selenite, and oxidation of seleno-dl-methionine to selenite existed in the metabolic pathways of the rat. We found that water-soluble selenium compounds led to a similar maximum content in blood and serum, but seleno-dl-methionine had a higher affinity for the brain and, by gel filtration chromatography, for the higher mol-wt (25-100 K Da) fractions of serum protein, when compared with inorganic forms.

Animals↗

Speciation analysis of arsenic and selenium compounds by capillary electrophoresis.

High-performance capillary electrophoresis is applied to the separation of different inorganic and organic arsenic and selenium compounds. In comparison with UV-detection, an approach with conductivity detection is described expecting higher sensitivity and universality. In this case the capillary was statically modified with CTAB before the electromigration procedure. The separation was performed with an electrolyte system consisting of CHES and Triton X-100. Detection limits of 0.06 mg/L or lower were obtained for As(V) and Se(VI). Water samples of an arsenic-polluted tailing of tin mining processes were analysed for anions as well as arsenic and selenium species.

Journal Article↗

Modulation of DNA repair processes by arsenic and selenium compounds.

Nickel, cadmium, cobalt and arsenic compounds are well known carcinogens to humans and experimental animals. In addition to the induction of mainly oxidative DNA damage, they interfere with nucleotide and base excision repair (BER) at low, non-cytotoxic concentrations. In case of arsenic, an inactivation of DNA repair has also been observed for the trivalent and pentavalent methylated metabolites, with the strongest effects exerted by MMA(III) and DMA(III). As potential molecular targets, interactions with so-called zinc finger proteins involved in DNA repair and/or DNA damage signaling have been identified. For example, arsenite suppresses poly(ADP-ribosyl)ation at extremely low, environmentally relevant concentrations. Also, Fpg and XPA involved in BER and NER, respectively, are inactivated by arsenite, MMA(III) and DMA(III). Nevertheless, an interaction with the zinc finger structures of DNA repair proteins may also occur by essential trace elements such as certain selenium compounds, which appear to exert anticarcinogenic properties at low concentrations but may compromise genetic stability at higher concentrations.

Animals↗

The impact of vitamins A,C,E, and selenium compound on prevention of liver cancer in rats.

A study was initiated to determine the impact of vitamins A, C, E, and selenium compound (Se) on the prevention of liver cancer. Sixty animals were fed a diet with or without these vitamins followed by aflatoxin B treatment for a period of 24 months. Most of the animals fed a diet devoid of vitamins developed liver cancer, while none or only a few of the animals given vitamins suffered during this period. We suggest that vitamins can inhibit liver cancer by inducing hepatic microsomal enzymes that metabolise aflatoxins to noncarcinogenic products.

Aflatoxin B1↗

Inhibition of DNA cytosine methyltransferase by chemopreventive selenium compounds, determined by an improved assay for DNA cytosine methyltransferase and DNA cytosine methylation.

The organoselenium compounds benzyl selenocyanate (BSC) and 1,4-phenylenebis(methylene)selenocyanate (p-XSC), as well as sodium selenite, are effective chemopreventive agents for various chemically induced tumors in animal models at both the initiation and postinitiation stages. The mechanisms involved at the postinitiation stage are not clear. Because several lines of evidence indicate that inhibition of excess DNA (cytosine-5)-methyltransferase (Mtase) may be a sufficient factor for the suppression or reversion of carcinogenesis, we examined the effects of sodium selenite, BSC, p-XSC and benzyl thiocyanate (BTC), the sulfur analog of BSC, on Mtase activity in nuclear extracts of human colon carcinomas, and of p-XSC on the Mtase activity of HCT116 human colon carcinoma cells in culture. For this purpose, we developed an improved Mtase assay, in which the incorporation of the methyl-[3H] group from S-adenosyl[methyl-3H]methionine into deoxycytidine of poly(dI-dC)-poly(dI-dC), is specifically determined by HPLC with radioflow detection after enzymatic hydrolysis, enhancing specificity and reliability. In a variation, using SssI methyltransferase and labeled S-adenosylmethionine, the overall methylation status of DNA in various tissues can also be compared. Selenite, BSC and p-XSC inhibited Mtase extracted from a human colon carcinoma with IC50s of 3.8, 8.1 and 5.2 microM, respectively; BTC had no effect. p-XSC also inhibited the Mtase activity and growth of human colon carcinoma HCT116 cells, with an IC50 of approximately 20 microM. The improved Mtase assay should prove to be a reliable method for screening potential Mtase inhibitors, especially using cells in culture. We suggest that inhibition of Mtase may be a major mechanism of chemoprevention by selenium compounds at the postinitiation stage of carcinogenesis.

Adenocarcinoma↗

Deactivation of singlet molecular oxygen by organo-selenium compounds exhibiting glutathione peroxidase activity and by sulfur-containing homologs.

The bimolecular rate constants (k) of quenching of molecular singlet oxygen 1O2 (1 delta g) by organo-selenium compounds exhibiting glutathione peroxidase activity and by sulfur analogs have been determined by time resolved phosphorescence detection of 1O2 in CD3OD and C6D6, with no solvent effect. The rate constants of quenching by the Se-containing compounds were found to be approximately one order of magnitude higher than those of the S-containing homologs. A linear correlation was observed between log k and the Hammett constant omega ortho with p = -0.89, the rate constant being higher for molecules with an electron-donating substituent and lower for those with an electron-withdrawing substituent. This observation is consistent with the involvement of a charge transfer complex in the deactivation of singlet oxygen.

Antioxidants↗

Determination of selenium compounds by HPLC with ICP-MS or FAAS as selenium-specific detector.

A speciation method was developed for selenious acid, selenic acid, trimethylselenonium ion (TMSe) and selenomethionine (SeMet). Separation of the four selenium species was achieved by HPLC on an ESA Anion III anion-exchange column using aqueous mobile phase of 5.5 mmol/L ammonium citrate at pH 5.5 with a flow rate of 1.5 mL/min. Under the optimal conditions, the four selenium species were separated within 8 minutes. On-line selenium-specific detection was carried out with an inductively coupled plasma mass spectrometer (ICP-MS) or a flame atomic absorption spectrometer (FAAS). The detection limits of HPLC-FAAS were approximately rho(Se) = 1 mg/L for each compound (100 microL injection). To increase the nebulization efficiency of the ICP-MS, the Meinhard concentric nebulizer was replaced by an ultrasonic nebulizer (USN). The ICP-MS signal intensity was increased by a factor of 7 for selenious acid and 24 to 31 for TMSe, SeMet and selenic acid with the USN compared to that with the Meinhard nebulizer. The detection limits of the HPLC-USN-ICP-MS were rho(Se) = 0.08 microgram/L for TMSe, rho(Se) = 0.34 microgram/L for selenious acid, rho(Se) = 0.18 microgram/L for SeMet and rho(Se) = 0.07 microgram/L for selenic acid.

Chromatography, High Pressure Liquid↗

Induction of retrovirus gene expression by selenium compounds.

Sodium selenite, sodium selenate, selenium oxide, selenophypoxanthine, selenopurine, selenocysteine, selenoethionine and selenomethionine were tested for their ability to induce endogenous retrovirus expression in cultured AKR mouse embryo fibroblasts. All except selenoethionine were highly toxic to the cells. Only selenomethionine however, had the ability to induce virus expression under the conditions used. The level of virus induction (plaque-forming-units/10(5) cells) was roughly proportional to dose over the range of concentrations from 0.25 mM to 5.0 mM. Induction was best observed when a treatment duration of 48 h was used and required the treatment of actively dividing cells. The induction and the cytotoxic effects of selenomethionine could be abrogated by simultaneous treatment with methionine. A ratio of methionine to selenomethionine of 1:10 inhibited induction by approx. 60% while equivalent amounts of methionine inhibited selenomethionine-mediated induction by greater than 96%, indicating that methionine was more efficiently recognized by the cells than was selenomethionine. A possible mechanism for selenomethionine induction involving the production of undermethylated DNA is presented.

Animals↗

Effects of organic and inorganic selenium compounds on rat mammary tumor cells.

To explore cellular effects of potent organoselenium chemopreventive agents we have used a rat mammary tumor cell line. We demonstrate that 1,4-phenylenebis(methylene) selenocyanate (p-XSC) at a dose of 5 microM is a more potent inhibitor of DNA, RNA and protein synthesis as well as of mitochondrial transmembrane potential than its chemopreventive counterparts benzyl selenocyanate (BSC) and sodium selenite. These differences were also reflected in reduced growth rate by 24 and 48 hr. Cell-cycle and cell-morphology analysis revealed that higher doses of p-XSC (10 microM) caused DNA fragmentation which was accompanied with partial loss of nuclear stainability, whereas BSC caused a noticeable change in cell-cycle distribution and extensive micronucleation. Overall, our results point to cellular targets of selenium compounds which may mediate their chemopreventive activities in mammary tissues.

Animals↗

Inhibition of human squalene monooxygenase by selenium compounds.

Selenosis in animals is characterized by a variety of neurological abnormalities, but the chemical species of selenium and the molecular targets that mediate this neurotoxicity are unknown. We have previously shown that selenite is a potent inhibitor of squalene monooxygenase, the second enzyme in the committed pathway for cholesterol biosynthesis; inhibition of this enzyme by dimethyltellurium leads to a peripheral demyelinating neuropathy similar to that seen in selenosis. To evaluate the role methylation plays in selenium toxicity, we examined the ability of three methylselenium compounds, methylselenol, dimethylselenide, and trimethylselenonium iodide, to inhibit purified recombinant human squalene monooxygenase. IC(50) values for methylselenol (95 microM) and dimethylselenide (680 microM) were greater than that previously obtained for selenite (37 microM), and inhibition by trimethylselenonium iodide was evident only at concentrations above 3 mM. Inhibition by methylselenol as well as by selenite was slow and irreversible, suggestive of covalent binding to the enzyme, and thiol-containing compounds could prevent and reverse this inhibition, indicating that these compounds were reacting with sulfhydryl groups on the protein. Monothiols such as glutathione and beta-mercaptoethanol provided better protection than did dithiols, suggesting that these selenium compounds bind to only one of the two proposed vicinal cysteines on squalene monooxygenase. Unexpectedly, the inhibition by selenite was significantly enhanced by dithiols, indicating that a more toxic species, possibly selenide, was formed in the presence of these dithiol reductants.

Animals↗

Glutathione peroxidase-like activity of simple selenium compounds. Peroxides and the heterocyclic N-oxide resazurin acting as O-atom donors.

Selenite and selenocystamine [(CyaSe)2] efficiently activate the decomposition of H2O2 by GSH and by other thiols, as demonstrated using a leuco crystal violet POD-based H2O2 assay which is applicable (unlike other assays) also in presence of thiols. The GPx-like activities were estimated to be 3.6 and 2.7 mumol H2O2/min per mumol SeO3(2-) and (CyaSe)2, respectively. Both selenium compounds also activate reduction of the heterocyclic N-oxide resazurin (RN-->O) to resorufin (RN) by GSH; H2O2 competes with reduction of this dye. GSSeH and CyaSeH, formed by interaction of GSH with SeO3(2-) and (CyaSe)2, respectively, are likely to be the active reductants. CyaSeH, generated gamma-radiolytically from (CyaSe)2, exhibits an absorption peak at 243 nm and is removed by H2O2 with a rate constant of 9.7 x 10(2) M-1 s-1, and slightly slower by hydroperoxides. We have no evidence for one-electron interactions between GSSeH or CyaSeH and H2O2, with formation of free radical intermediates, as previously proposed in the case of selenium-activated reduction of cytochrome c by GSH (Levander et al., Biochemistry 23, 4591-4595 (1973)). Our results can be explained by O-atom transfer from the substrate to the active selenol group, RSeH + H2O2 (RN-->O)-->RSeOH + H2O (RN), and recycling of RSeOH to RSeH (+ H2O) by GSH, analogous to the selenenic acid pathway of GPx. The substrate specificity appears to be different, however, in that GPx is unable to catalyse RN-->O reduction, and GSSeH hardly catalyses the decomposition of cumene- or t-butyl-hydroperoxide; CyaSeH, on the other hand, is active also with the hydroperoxides. RN-->O is reduced to RN also by certain oxidizing free radicals, e.g. by the thiyl CyaS.., O-atom transfer may in this case lead to the generation of reactive oxyl radicals.

Catalysis↗