Selenium uptake by Butyrivibrio fibrisolvens and Bacteroides ruminicola

Abstract The uptake and incorporation of ⁷⁵[Se]selenite by Butyrivibrio fibrisolvens and Bacteroides ruminicola were by constitutive systems. Rates of uptake were higher in chemostat culture than in batch culture and there may be some inducible component. Uptake of [⁷⁵Se]selenite was distinct from sulphate or selenate transport, since sulphate and selenate did not inhibit selenite uptake, nor could sulphate or selenate uptake be demonstrated in these organisms. Selenite uptake in B. fibrisolvens had and apparent K _m of 1.74 mM and a V _max of 109 ng Se · min⁻¹· (mg protein)⁻¹. An apparent K _m of 1.76 mM and V _max of 1.5 μg Se · min⁻¹· (mg protein)⁻¹ was obtained for B. ruminicola . [⁷⁵Se]Selenite uptake by both organisms was partially sensitive to inhibition by 2,4-DNP. Uptake by B. fibrisolvens was also partially inhibited by azide and arsenate and in B. ruminicola it was partially inhibited by fluoride. CCCP, CPZ, DCCD or quinine did not inhibit uptake in either B. fibrisolvens or B. ruminicola . Selenite transport by both organisms was sensitive to IAA and NEM and was strongly inhibited by sulphite and nitrite. [⁷⁵Se]Selenite was converted to selenocystine, selenohomocystine and selenomethionine by B. fibrisolvens. B. ruminicola did not incorporate [⁷⁵Se]selenite into organic compounds, but did reduce it to red elemental selenium.     

Selenium modifies carcinogen metabolism by inhibiting enzyme induction

Since selenium has been found to exert a protective action against carcinogenesis in various systems, the mechanism where-by sodium selenite inhibits DNA binding of the carcinogen, 7,12-dimethylbenz[a]anthracene, was investigated. It was found that selenite preferentially reduced DNA binding occurring through ananti-dihydrodiol epoxide metabolite of this carcinogen by inhibiting the induction of an enzyme system that generates this specific reactive metabolite.     

Culture duration alters the glutathione content and sensitivity to ethacrynic acid of rat hepatocyte monolayer cultures

Many of the differentiated functions of hepatocytes are lost in culture, yet addition of certain medium supplements can aid in the retention of differentiated character. Therefore, the effect of time in monolayer culture on rat hepatocyte glutathione (GSH) synthesis and sensitivity to the GSH detoxicated xenobiotic ethacrynic acid was examined in cultures with and without medium supplementation by transferrin and sodium selenite. GSH content was found to be about 12 nmol/µg DNA at 4 hr in culture and to approximately triple by 24 hr. Intracellular GSH levels continued to increase in transferrin/sodium selenite-supplemented cultures, from 32 to 41.6 nmol/µg DNA, while GSH levels in unsupplemented cultures declined to 18 nmol/µg DNA. However, the rate of GSH synthesis after diethylmaleate depletion was found to decrease from 4.2 to 2.8 nmol/hr/µg DNA at 4 and 24 hr after inoculation, respectively. GSH repletion rate increased to 3.9 nmol/hr/µg DNA at 48 hr. The GSH accumulation rate after depletion in supplemented cultures did not vary significantly over the initial 48 hr. Incubation for 3 hr with 100 µM ethacrynic acid (EA) did not elicit an increase in LDH leakage in hepatocyte monolayers after 4 or 48 hr in culture or in cultures with supplemented medium at any time point tested. Cultures 24 hr in medium without transferrin/sodium selenite supplementation exhibited significant LDH leakage after 3 hr of EA treatment. Over the 3 hr EA treatment, intracellular GSH content was decreased in all cultures. Only in the 24 hr unsupplemented cultures did GSH depletion exceed the 90% level previously associated with depletion of the mitochondrial pool of GSH and EA toxicity in hepatocytes. The experiments show that during the redifferentiation of hepatocytes in culture, a transient period occurs when apparent GSH synthesis is depressed and enhanced sensitivity to GSH-detoxicated compounds is observed. This period of increased sensitivity is prevented or at least delayed by inclusion of supplemental transferrin and sodium selenite, suggesting that redifferentiation can be regulated by extracellular influences.Abbreviations CYSSG cysteine-glutathione mixed disulfide - DEM diethyl maleate - EA ethacrynic acid - GSH reduced glutathione - GSSG oxidized glutathione - HBS HEPES buffered saline - HWME hepatocyte Williams' Medium E (WME with insulin, corticosterone and 0.5 mM methionine) - LDH lactate dehydrogenase - TS-HWME transferrin/sodium selenite-supplemented HWME - WME Williams' Medium E     

Enhancement of the repair of carcinogen-induced DNA damage in the hamster pancreas by dietary selenium

We measured single strand breaks (SSB) in pancreas DNA produced by N-nitrosobis(2-oxopropyl)amine (BOP) in hamsters fed purified diets containing added sodium selenite (Se) at 0.0, 0.1 and 5.0 ppm. There were fewer SSB in those given the 5.0 ppm Se diet throughout the experiment. One hour after dosing with BOP (20 mg/kg), there were 2.26 ± 0.47, 2.83 ± 0.43 and 1.74 ± 0.43 SSB per 10⁸ daltons (mean ± S.E.M.) respectively in the three groups. The SSB were repaired faster in the 5.0 ppm Se-fed group. The approximate half-lives of the SSB were 33, 30 and 8 days, respectively. In the hamsters fed 5.0 ppm Se there was a small, statistically significant increase in pancreatic DNA synthesis. Autoradiographic analysis indicated that this was repair synthesis. In a second experiment, hamsters were fed one of the three diets prior to and for 2 days after administration of a single dose of BOP (20 mg/kg). They were then fed the 5.0 ppm Se diet for 5 days. The number of SSB was compared with those in hamsters fed their original diet for 7 days after BOP dosing. There was a statistically significant difference in the number of SSB in the hamsters fed 0.1 ppm Se before and for 2 days after BOP. In these hamsters there were 1.21 ± 0.24 SSB per 10⁸ daltons compared with 3.19 ± 0.4 (mean ± S.E.M.). These results suggest high levels of dietary Se stimulate the repair of carcinogen-induced DNA damage.     

Effects of glutathione and of cysteine on intestinal absorption of selenium from selenite

The influence of glutathione (1 mmol/L) (GSH) on in vitro mucosal uptake and in vivo absorption of⁷⁵Se-labeled selenite (10 μmol/L) was investigated in rat jejunum. For comparison, the effect ofl-cysteine (1 mmol/L) on in vivo absorption of⁷⁵Se-labeled selenite was also studied. In the in vitro, uptake experiments, only the mucosal surface was exposed to the incubation medium for 3 min. For the in vivo experiments, a luminal perfusion technique was employed. GSH inhibited in vitro mucosal Se uptake, whereas absorption in vivo was stimulated by GSH.l-Cysteine also stimulated in vivo Se absorption, confirming former in vitro mucosal uptake experiments. Thus, unlikel-cysteine, GSH affected in vitro and in vivo absorption of Se from selenite differently. Enzymatic cleavage of products of the reaction of selenite with GSH occuring more efficiently under in vivo than in vitro conditions may be a prerequisite for the stimulatory effect of GSH on Se absorption. This apparently does not apply to the stimulatory effect of cysteine. Since, GSH occurs in the intestinal lumen under physiological conditions, it may contribute to the high bioavailability of Se from selenite.     

Stimulation of mucosal uptake of selenium from selenite by some thiols at various sites of rat intestine

The influence of several thiols (conc. 1 mmol/L) on mucosal uptake of⁷⁵Se from⁷⁵Se-labeled selenite (conc. 10 μmol/L) across the brush border of rat jejunum and cecum was investigated in vitro using a short-term uptake technique.l-Cysteine (l-Cys) stimulated⁷⁵Se uptake in the mid- and distal jejunum and cecum, but not in the proximal jejunum. The effect was maximal in the distal jejunum.d-Cys was less effective in the jejunum and similarly effective in the cecum.l-Leucine (l-Leu) andl-glutamic acid significantly reduced the stimulatory effect ofl-Cys on Se uptake in the distal jejunum, whereas the respective effect ofd-Cys was not diminished byl-Leu. Cysteamine stimulated mucosal⁷⁵Se uptake at all intestinal sites tested, whereas the effect of mercaptopyruvate was restricted to the distal jejunum. Thioglycolate also enhanced⁷⁵Se uptake in the distal jejunum. The stimulatory effects ofl-Cys, mercaptopyruvate, and thiologlycolate were Na⁺-dependent, whereas the effect of cysteamine also occurred in the absence of Na⁺. Mercaptosuccinate,d-penicillamine, ergothioneine, and thiosulfate did not enhance mucosa⁷⁵Se uptake. It is concluded from these findings that the reaction of some thiols with selenite results in Se compounds that are rapidly absorbed by the intestinal epithelium through various Na⁺-dependent and Na⁺-independent, mechanisms. The high bioavailability of Se from selenite found by others might thus be the result of the presence of thiols in the gastrointestinal tract.     

The antimutagenic effect of selenium on 7,12-dimethylbenz(a)anthracene and metabolites in the amesSalmonella/microsome system

The antimutagenic effect of selenium as sodium selenite, sodium selenate, selenium dioxide, and seleno-methionine was studied in the AmesSalmonella/microsome mutagenicity test using 7,12-dimethylbenz(a)anthracene (DMBA) and some of its metabolites. Selenium (20 ppm) as sodium selenite reduced the number of histidine revertants on plates containing up to 100 μg DMBA/plate. Increasing concentrations of selenium as sodium selenite, sodium selenate, and selenium dioxide up to 40 ppm Se progressively decreased the number of revertants caused by 50 μg DMBA. DMBA and its metabolites 7-hydroxymethyl-12-methylbenz(a)anthracene, 12-hydroxymethyl-7-methylbenz(a)anthracene, and 3-hydroxy-7,12-dimethylbenz(a)anthracene were mutagenic forSalmonella typhimurium TA100 in the presence of an S-9 mixture. Selenium supplementation as Na₂SeO₃ reduced the number of revertants induced by these metabolites to background levels. The antimutagenic effect of inorganic selenium compounds cannot be explained by toxicity of selenium as determined by viability tests withSalmonella typhimurium TA100. Selenium supplementation in all forms examined, except sodium selenate, decreased the rate of spontaneous reversion. Selenium as sodium selenate was slightly mutagenic at concentrations of 4 ppm or less. Higher concentration of Na₂SeO₄ inhibited the mutagenicity of DMBA. The present studies support the anticarcinogenic potential of selenium and indicate that form and concentration are important factors in this trace element's efficacy.     

Application of sodium selenate to cowpea (Vigna unguiculata L.) increases shoot and grain Se partitioning with strong genotypic interactions

BackgorundCowpea is a crop widely used in developing countries due its rusticity. Besides its rich genotypic variability, most breeding programs do not explore its potential to improve elements uptake. Selenium (Se) is a scarce element in most soils, resulting in its deficiency being common in human diets. This study aimed to evaluate the interaction between biofortification with Se and genotypic variation in cowpea, on the concentrations of Se in roots, leaves + stem and grains.MethodsTwenty-nine cowpea genotypes were grown in a greenhouse in the absence (control) and presence of Se (12.5 μg Se kg⁻¹ soil) as sodium selenate, in fully randomized scheme. The plants were cultivated until grains harvest. The following variables were determined: roots dry weight (g), leaves + stems dry weight (g), grains dry weight (g), Se concentration (mg kg⁻¹) in roots, leaves + stems and grains, and Se partitioning to shoots and grains.ResultsSelenium application increased the Se concentration in roots, leaves + stems and grains in all genotypes. At least twofold variation in grain Se concentration was observed among genotypes. Selenium application did not impair biomass accumulation, including grain dry weight. Genotype “BRS Guariba” had the largest Se concentration in grains and leaves + stems. Genotype MNC04-795 F-158 had the largest partitioning of Se to shoots and grain, due to elevated dry weights of leaves + stems and grain, and high Se concentrations in these tissues.ConclusionThis information might be valuable in future breeding programs to select for genotypes with better abilities to accumulate Se in grain to reduce widespread human Se undernutrition.     

Effects of Na2SeO3 on growth,metabolism, antioxidase and enzymes involved in polysaccharide synthesis of Cordyceps militaris

The main objective of our study was to illuminate effects of sodium selenite (Na₂SeO₃) on growth, metabolism and enzyme activities of Cordyceps militaris. By adding Na₂SeO₃ in fermentation medium (7 mg/L), it was found that mycelium of C. militaris was stronger with more plump spores and its biomass was higher accompanied by the maximum (13.19 g/L) at 6 d of culture. Besides, Na₂SeO₃ also caused the enhancement of total thiol (T-SH), non-protein thiol (NP-SH) contents and the biosynthesis of Se-polysaccharide (Se-CMP) with discriminatory molecular weight, optical rotation, UV–vis and FT-IR spectra. Activities of antioxidase including catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and enzymes involved in polysaccharide synthesis including phosphoglucomutase (PGM), phosphoglucose isomerase (PGI), UDPG-pyrophosphorylase (UGP) enhanced to different extent. Their corresponding genes were also up-regulated but cat gene (encoding CAT). Se-enrichment of C. militaris provided a good way for desirable biomass and polysaccharide biosynthesis in further research.