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The purification, crystallization and preliminary structural characterization of human MAWDBP, a member of the phenazine biosynthesis-like protein family.

MAWDBP is the only representative of the phenazine biosynthesis-like protein family in the human genome. Its expression is elevated in several disease processes, including insulin resistance, folate deficiency and hypotension, and it may also be involved in carcinogenesis. The exact molecular function of MAWDBP is unknown. Native and seleno-L-methionine-labelled MAWDBP were expressed in Escherichia coli and crystallized at room temperature from precipitants containing 10 mM KF, 14%(w/v) PEG 3350 and 0.1 M sodium citrate pH 5.4. Crystals belong to space group H32, with unit-cell parameters a = b = 187, c = 241 A, indicative of three to five monomers per asymmetric unit. Crystals were cryoprotected with 15%(v/v) glycerol and data have been collected to 2.7 A resolution.

Cloning, Molecular↗

Chemoprevention of 7,12-dimethylbenz[a]anthracene-induced mammary carcinogenesis in rat by the combined actions of selenium, magnesium, ascorbic acid and retinyl acetate.

The chemopreventive actions of sodium selenite (SS), magnesium chloride (MC), ascorbic acid (AA) and retinyl acetate (RA), given singly or in combinations, on mammary carcinogenesis induced by 30 mg of 7,12-dimethylbenz[a]anthracene (DMBA) in female adult rats were evaluated. Administration of modulators was carried out from the age of 40 +/- 3 days to 240 +/- 3 days. When DMBA alone was given 100% of the rats developed mammary tumors. When modulators were given singly the tumor incidences were reduced to 51.77% (SS), 46.4% (MC), 57.1% (AA) and 48.1% (RA). When the modulators were given in combination of twos, the tumor incidences were further reduced to 29.5% (SS + MC), 31% (SS + AA), 29.6% (SS + RA), 25.9% (MC + AA), 31.8% (MC + RA) and 34.6% (AA + RA). Administration of modulators in combinations of threes resulted in still further reduction of tumor incidences to 22.2% (SS + MC + AA), 19.2% (SS + MC + RA), 16% (MC + AA + RA) and 23.1% (AA + RA + SS). When all four modulators were given concurrently the tumor incidence was only 12%. Further, the number of tumors per tumor-bearing animal declined with the increase in the number of agents used in combination for modulation.

9,10-Dimethyl-1,2-benzanthracene↗

Comparative effects of selenite and selenate on nitrate assimilation in barley seedlings.

The effect of SeO3= and SeO4= on NO3- assimilation in 8-d-old barley (Hordeum vulgare L.) seedlings was studied over a 24-h period. Selenite at 0.1 mol m-3 in the uptake solutions severely inhibited the induction of NO3- uptake and active nitrate reductases. Selenate, at 1.0 mol m-3 in the nutrient solution, had little effect on induction of activities of these systems until after 12 h; however, when the seedlings were pretreated with 1.0 mol m-3 SeO4= for 24 h, subsequent NO3- uptake from SeO4(=) -free solutions was inhibited about 60%. Sulphate partially alleviated the inhibitory effect of SeO3= when supplied together in the ambient solutions, but had no effect in seedlings pretreated with SeO3=. By contrast, SO4= partially alleviated the inhibitory effect of SeO4= even in seedlings pretreated with SeO4=. Since uptake of NO3- by intact seedlings was also inhibited by SO3=, the percentage of the absorbed NO3- that was reduced was not affected. By contrast, SeO4=, which affected NO3- uptake much less, inhibited the percentage reduced of that absorbed. However, when supplied to detached leaves, both SeO3= and SeO4= inhibited the in vivo reduction of NO3- as well as induction of nitrate reductase and nitrite reductase activities. Selenite was more inhibitory than SeO4= ; approximately a five to 10 times higher concentration of SeO4= than SeO3= was required to achieve similar inhibition. In detached leaves, the inhibitory effect of both SeO3= and SeO4= on in vivo NO3- reduction as well as on the induction of nitrate reductase activity was partially alleviated by SO4=. The inhibitory effects of Se salts on the induction of the nitrite reductase were, however, completely alleviated by SO4=. The results show that in barley seedlings SeO3= is more toxic than SeO4=. The reduction of SeO4= to SeO3= may be a rate limiting step in causing Se toxicity.

Biological Transport↗

Confluent and reticulated papillomatosis responsive to selenium sulfide.

Confluent and reticulated papillomatosis of Gougerot and Carteaud (CRP) is a distinctive clinicopathologic entity, of unknown etiology, which shows some resemblance to pityriasis versicolor. A case of CRP that showed a significant therapeutic response to topical selenium sulfide is reported. In previously described cases, response to this agent and to other medications, both antifungal and keratolytic, has been variable. A review of these cases reveals conflicting evidence regarding the questionable role of Malassezia furfur in CRP.

Administration, Topical↗

Selenium binding to beef-kidney rhodanese.

The reaction of beef kidney rhodanese with selenosulfate was studied. The selenium-treated enzyme shows an absorption spectrum with a maximum at 375 nm attributable to a sulfoselenide group. This absorption is bleached by addition of cyanide. After cyanide treatment stoichiometric amount of selenocyanate can be found. The intrinsic fluorescence of rhodanese is quenched by addition of stoichiometric selenosulfate. This effect can be reversed by cyanide or sulfite but not by selenite or glutathione. By comparison with model complexes the selenium-rhodanese intermediate was identified as a cysteinyl-selenium derivative.

Animals↗

Differences in the binding of sulfate, selenate and thiosulfate ions to bovine liver rhodanese, and a description of a binding site for ammonium and sodium ions. An X-ray diffraction study.

The binding of sulfate, selenate and thiosulfate by the sulfur-transferase rhodanese (EC 2.8.1.1) in the crystalline state has been studied by X-ray analysis at resolutions between 0.23 nm and 0.4 nm. The three ions appear to occupy a common site between the N eta atoms of Arg-29 and the main-chain NH group of Glu-148 at the surface of the enzyme molecule. A second binding site for the three ions is situated at the entrance to the active centre, between the side chains of Arg-186 and Lys-249. Selenate and thiosulfate are bound equally well at both anion-binding sites. Sulfate, however, binds better at the first position, near Arg-29, than at the second site near Arg-186. In the complex of sulfur-rhodanese with thiosulfate, the outer sulfur atom of the anion near the active centre points towards the extra sulfur atom which is bound as a persulfide to the S gamma of the essential Cys-247. The distance between the outer sulfur atom of the thiosulfate ion and the persulfide sulfur atom appears to be about 0.3 nm. The thiosulfate difference Fourier also shows a distinct, localized conformational change involving residues 71, 72 and 249. This is the result of the replacement of an ammonium ion in the sulfate and selenate media by a sodium ion in the sodium thiosulfate solution. Rhodanese is apparently able to accomodate ions with different radii at this cation-binding site by minor structural alterations.

Ammonium Sulfate↗

Effects of a supranutritional dose of selenate compared with selenite on insulin sensitivity in type II diabetic dbdb mice.

The present study was performed to examine the mechanism by which selenate ameliorates the insulin sensitivity in type II diabetic dbdb mice. Therefore, 21-adult female dbdb mice were randomly assigned to three experimental groups (0Se, SeIV and SeVI) with seven animals per group. Mice of group 0Se were fed with a selenium-deficient diet (<0.02 mg Se/kg) based on wheat and torula yeast for 8 weeks whereas the mice of groups SeIV (selenite) and SeVI (selenate) were fed with sodium selenite and sodium selenate (up to 35% of the LD(50) for mice in eighth week), in addition to the diet by daily tube feeding. Eight weeks of selenate application led to significantly elevated insulin sensitivity in comparison with selenium deficiency and selenite application. The activity of cytosolic protein tyrosine phosphatases (PTPs) as important negative regulators of insulin signalling was reduced from 53.8% to 22.5% in the liver and skeletal muscle of selenate-treated mice in comparison with the selenium deficient and selenite-treated controls, suggesting an inhibition of PTPs by intermediary selenate metabolites. In an additional in vitro inhibition study, selenate (oxidation state +VI) did not inhibit PTP activity. Selenium metabolites in the oxidation state +IV were found to be the actual inhibitors of PTP activity. In conclusion, the results of the present study show that one possible mechanism by which supranutritional selenate doses enhance insulin sensitivity in type II diabetic dbdb mice is based on the inhibition of PTPS as negative regulators of insulin signalling. Moreover the cellular metabolism of selenate including its intermediary reduction to the oxidation state +IV seems to play a crucial role during this process.

Animals↗

Removal of selenate from sulfate-containing media by sulfate-reducing bacterial biofilms.

A biofilm-selected strain of a Desulfomicrobium sp. removed selenate from solution to sub-micromolar concentrations during growth on lactate (or hydrogen) and sulfate. Under sulfate-limited growth conditions, selenium was enzymatically reduced to selenide. Under excess sulfate conditions, selenate removal was primarily by enzymatic reduction to elemental selenium. Sequestration by biofilms was greater under the latter condition. Experiments with washed cell suspensions showed that high sulfate concentrations inhibited cell-specific selenate reduction, but when growing cells were exposed to selenate, the biomass increase achieved during incubations with abundant sulfate resulted in more rapid selenate removal. The addition of small amounts of sulfite, or thiosulfate, ameliorated this inhibition. Nitrate also inhibited selenate reduction in washed cell suspensions, apparently due to a general oxidizing effect. These results suggest that where biofilm-based sulfate-reducing bacteria (SRB) bioreactors are considered for the treatment of mixed metalliferous wastes that contain selenium oxyanions, adequate selenate removal should be achievable under a range of environmental conditions. The form and fate of the precipitated product will, however, be influenced by the dominant reduction pathway, which is controlled by environmental variables.

Biodegradation, Environmental↗

Mapping quantitative trait loci associated with selenate tolerance in Arabidopsis thaliana.

Selenium is essential for many organisms, but is toxic at higher levels. To investigate the genetic basis of selenate tolerance in Arabidopsis thaliana, quantitative trait loci (QTL) associated with selenate tolerance in accessions Landsberg erecta and Columbia were mapped using recombinant inbred lines (RILs). The selenate tolerance index (TI(D10) = root growth + 30 microm selenate/root growth control x 100%) was fourfold higher for parental line Col-4 (59%) than for parent Ler-0 (15%). Among the 96 F8 RILs, TI(D10) ranged from 11 to 75% (mean 37%). Using composite interval mapping, three QTL were found on chromosomes 1, 3 and 5, which together explained 24% of variation in TI(D10) and 32% of the phenotypic variation for the difference in root length +/- Se (RL(D10)). Highly significant epistatic interactions between the QTL and markers on chromosome 2 explained additional variation for both traits. Potential candidate genes for Se tolerance in each of the QTL regions are discussed. These results offer insight into the genetic basis of selenate tolerance, and may be useful for identification of selenate-tolerance genes.

Arabidopsis↗

Confluent and reticulated papillomatosis associated with tinea versicolor in three siblings.

We describe three teenage siblings with confluent and reticulated papillomatosis, all presenting during a 6-month period. Two of the three patients had confirmed tinea versicolor, with positive potassium hydroxide scrapings, in association with this entity. This is the largest series of siblings with confluent and reticulated papillomatosis, and the only report describing family members having both confluent and reticulated papillomatosis and tinea versicolor. This report lends further evidence to the hypothesis that confluent and reticulated papillomatosis may be etiologically linked to tinea versicolor, and also suggests a genetic predisposition for it.

Adolescent↗

Acetate, lactate, propionate, and isobutyrate as electron donors for iron and sulfate reduction in Arctic marine sediments, Svalbard.

The contribution of volatile fatty acids (VFA) as e(-)-donors for anaerobic terminal oxidation of organic carbon through iron and sulfate reduction was studied in Arctic fjord sediment. Dissolved inorganic carbon, Fe(2+), VFA concentrations, and sulfate reduction were monitored in slurries from the oxidized (0-2 cm) and the reduced (5-9 cm) zone. In the 0-2 cm layer, 2/3 of the mineralization could be attributed to sulfate reduction and 1/3 to iron reduction. In the 5-9 cm layer, sulfate reduction was the sole mineralization process. Acetate and lactate turnover rates were measured by radiotracer. Inhibition of sulfate reduction with selenate resulted in the accumulation of acetate, propionate, and isobutyrate. The acetate turnover rates determined by radiotracer and accumulation after inhibition were similar. VFA turnover accounted for 21% and 52% of the mineralization through sulfate reduction in the 0-2 and 5-9 cm layer, respectively. Acetate and lactate turnover in the inhibited 0-2 cm slurry was attributed to iron reduction and accounted for 10% and 2% of the iron reduction. Therefore, 88% and 79% of the iron and sulfate reduction in the 0-2 cm layer, respectively, must be fueled by alternative e(-)-donors. The accumulation of VFA in the selenate-inhibited 0-2 cm slurry did not enhance iron reduction, indicating that iron reducers were not limited by VFA availability.

Acetates↗