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A selenium-dependent and a selenium-independent formylmethanofuran dehydrogenase and their transcriptional regulation in the hyperthermophilic Methanopyrus kandleri.

The genome of Methanopyrus kandleri was found to harbour a gene, fwuB, predicted to encode the catalytic subunit of a tungsten formylmethanofuran dehydrogenase with an active site selenocysteine, and a second gene, fwcB, encoding a tungsten formylmethanofuran dehydrogenase with an active site cysteine. Northern blot and primer-extension analysis revealed that both genes were differentially transcribed. During growth of the methanogen on medium supplemented with selenium only fwuB was transcribed, whereas transcription of both fwuB and fwcB was observed on selenium-deprived medium. Growth of M. kandleri was stimulated by tungstate and selenite but not by molybdate. The findings indicate that the hyperthermophilic archaeon contains two tungsten isoenzymes of formylmethanofuran dehydrogenase, one of which is a novel selenium enzyme. They also indicate that the hyperthermophilic methanogen probably does not contain a molybdenum formylmethanofuran dehydrogenase which appears to be present only in thermophilic and mesophilic methanogens.

Aldehyde Oxidoreductases↗

Synthesis of 5-methylaminomethyl-2-selenouridine in tRNAs: 31P NMR studies show the labile selenium donor synthesized by the selD gene product contains selenium bonded to phosphorus.

An enzyme preparation from Salmonella typhimurium catalyzes the conversion of 5-methylaminomethyl-2-thiouridine in tRNAs to 5-methylaminomethyl-2-selenouridine when supplemented with selenide and ATP. Similar preparations from a Salmonella mutant strain carrying a defective selD gene fail to catalyze this selenium substitution reaction. However, supplementation of the deficient enzyme preparation with the purified selD gene product (SELD protein) restored synthesis of seleno-tRNAs. In the absence of the complementary enzyme(s), the SELD protein catalyzes the synthesis of a labile selenium donor compound from selenide and ATP. 31P NMR studies show that among the products of this reaction are AMP and a compound containing selenium bonded to phosphorus. The reaction is completely dependent on the addition of both selenide and magnesium. The dependence of reaction velocity on ATP concentration shows sigmoidal kinetics, whereas dependence on selenide concentration obeys Michaelis-Menten kinetics indicating a Km value of 46 microM for selenide.

Adenosine Monophosphate↗

Selenium species determination in selenium-enriched pumpkin (Cucurbita pepo L.) seeds by HPLC-UV-HG-AFS.

Pumpkins were treated by spraying the leaves in the flowering period with a water solution containing 1.5 mg Se per liter in the form of Na2SeO4. The average total selenium content of seeds was found to be 0.19 microg g(-1) in nontreated pumpkins and 1.1 microg g(-1) in exposed ones. For speciation analysis, enzymatic hydrolysis with different amounts of Protease XIV was carried out. Under optimal conditions of enzymatic hydrolysis, 90% of the total selenium was found in soluble forms. Separation of species was performed using HPLC on anion and cation exchange columns and for detection UVHG-AFS was applied. In enzymatic hydrolysis extracts, the main fraction of selenium was bound as selenomethionine (SeMet), representing on average of 81 +/- 8% of the total Se content in the sample.

Chromatography, High Pressure Liquid↗

Effect of supranutritional and organically bound selenium on performance, carcass characteristics, and selenium distribution in finishing beef steers.

Dietary selenium influences the Se content in edible muscle of beef cattle. Limited data are available to describe the effects that feeds naturally high in Se have on production, carcass characteristics, and Se distribution in terminal tissues. Therefore, 43 crossbred steers (BW = 351 +/- 24 kg) were stratified by BW and assigned to one of four dietary treatments: Se adequate (CON; n = 12), Se provided as high-Se wheat (WHT; n = 9), high-Se hay (HAY; n = 11), or sodium selenate (SEO; n = 11). Daily selenium intake for WHT, HAY, and SEO diets was 65 microg/kg BW, whereas it was 9.5 microg/kg BW for CON. Diets were similar in ingredient composition (25% wheat, 39% corn, 25% grass hay, 5% desugared molasses, and 6% wheat middling-based supplement; DM basis), isonitrogenous and isocaloric (14.0% CP, 2.12 Mcal NEm/kg DM and 1.26 Mcal NEg/ kg DM), and offered once daily (1500) individually to steers in a Calan gate system for 126 d. At the end of the trial, steers were slaughtered; carcass data were recorded; and samples of the liver, kidney, spleen, semitendinosus, and hair were collected for Se analysis. Intake of DM, G:F, and ADG did not differ (P > 0.13). No differences (P > 0.12) were noted for hot carcass weight, organ weights, longissimus muscle area, back-fat thickness, marbling scores, or quality and yield grade. Kidney, pelvic, and heart fat tended to be higher (P = 0.06) in CON and WHT compared with SEO and HAY steers (2.9, 2.4, 2.5, 2.9 +/- 0.2% for CON, SEO, HAY, and WHT, respectively). Selenium concentrations in all tissues collected differed (P < 0.003) due to treatment. Distribution of Se to the kidney, spleen, and hair were similar with CON < SEO < HAY < WHT (8.40, 10.05, 10.86, 12.89 +/- 0.26 ppm for kidney; 2.00, 2.60, 3.82, 5.16 +/- 0.09 ppm for spleen; 1.80, 4.00, 5.93, 10.54 +/- 0.56 ppm for hair; P < 0.01). The distribution of Se in liver and muscle (DM basis) differed from that in other tissues, with CON < HAY < SEO = WHT (2.33, 6.56, 9.91, 10.79 +/- 0.80 ppm; P < 0.01) and CON = SEO < HAY < WHT (1.33, 1.55, 3.32, 4.41 +/- 0.18 ppm; P < 0.01), respectively. When providing dietary Se at supranutritional levels, source of Se did not affect production or carcass characteristics, but it altered the distribution and concentration of Se throughout the tissues of finishing beef steers.

Animal Feed↗

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↗

[Determination of selenium in selenium-enriched foods by hydride generation atomic absorption spectrometry].

OBJECTIVE: To determine selenium (Se) in selenium-enriched foods by hydride generation atomic absorption spectrometry. METHODS: The samples were digested by HNO3-HCLO4 and reduced by NaHB4, and then was determined in 6 mol/L HCL. RESULTS: The results showed that the detectable range, detection limit, recovery rate, and the relative standard deviation(RSD) of the method were 0 - 80.0 microg/L, 0.0056 mg/kg, 92.05% - 103.88%, and 1.34% - 3.81% respectively. CONCLUSION: This method is suitable for selenium determination.

Agaricales↗

Selenium bioavailability as selenite (74Se) and as a selenium drug (76Se) by stable isotope methodology.

Many forms of selenium supplement are commercialized. The purpose of this work was to study the bioavailability of one selenium drug, "Granions de Sélénium", as compared with sodium selenite. Each product was labelled with enriched stable isotopes, and by means of gas chromatography/mass spectrometry (GC/MS), the metabolism of the two forms was followed simultaneously. In a first study, the absorption of a single dose of 480 micrograms Se (Na-Selenite) containing 100 micrograms of 74Se was compared with that of 480 micrograms Se (Granions) containing 100 micrograms of 76Se. Measurement precision was mediocre and only apparent absorption, urinary excretion and body retention could be measured. In a second experiment, kinetic studies were performed after a single oral dose of 100 micrograms of 74Se (Na-selenite) and 100 micrograms of 76Se (Granions). Blood, urine and feces samples were collected for a week to determine the true absorption, plasma half-life, and turnover of these two compounds. No significant difference in the metabolism of selenium was observed between "Granions de Sélénium" and selenite. Both present two maximal absorption peaks at 6 h and 12 h and a bioavailability of 76%.

Adult↗

Chemical forms of selenium in corn and rice grown in a high selenium area of China.

The chemical forms of selenium were determined in maize grown near Yutangba Village and in rice grown near Haubei village, Exi Prefecture, Enshi county of Hubei Province, China. The maize sample contained 18 ppm and the rice samples an average of 3.6 ppm selenium. After they were ground to obtain a fine flour, this was acid hydrolyzed with hydrochloric acid in an inert atmosphere and the hydrolyzates chromatographed on columns of Dionex DC6A resin. The results indicate that the majority of the selenium is present as selenomethionine in both rice and corn.

China↗

Induction of Escherichia coli mastitis in cows fed selenium-deficient or selenium-supplemented diets.

Ten Holstein heifers were fed a selenium-deficient (SeD) diet (0.04 mg of Se/kg on a total ration dry-matter basis) 3 months before calving and throughout their first lactation. A selenium-supplemented (SeS) diet (2 mg of Se/head/d) was fed to a group of 10 heifers. In about the 14th week of lactation, the cows were challenge-exposed to Escherichia coli by administering 15 to 40 colony-forming units (CFU) into 1 mammary gland. Selenium concentration (microgram/ml) in blood around the time of challenge exposure was 0.033 +/- 0.002 (mean +/- SEM) in SeD and 0.132 +/- 0.006 in SeS cows. Infections were established in all challenge-exposed quarters. The frequency of quarter atrophy and agalactia, and reduction in whole-udder milk yield in the first 4 days after challenge exposure, were greater (P less than 0.05) in the SeD cows. Log10 peak bacterial concentrations in milk were higher (P less than 0.05) in SeD (7.63 +/- 0.34 CFU/ml) than in SeS cows (5.57 +/- 0.66 CFU/ml). Mean log bacterial concentration was significantly higher (P less than 0.05) from 12 to 20 hours after challenge exposure in SeD than in SeS cows. Duration of infection was significantly greater (P less than 0.05) in SeD (162.0 +/- 12.0) than in SeS cows (114.4 +/- 18.0 hours). Milk somatic cell counts increased significantly more slowly (P less than 0.05) in SeD than in SeS cows from 8 to 16 hours after challenge exposure. Ratios of milk somatic cells to bacteria in milk were significantly lower (P less than 0.05) in SeD than in SeS cows at 12 and 16 hours after challenge exposure.

Animals↗

The absence of adaptive changes in tissue activities of glutathione-S-transferase, superoxide dismutase and catalase following selenium and vitamin E supplementation in subjects with low selenium status.

Three groups of New Zealand women were given daily in a double blind randomised study, 200 micrograms Se as sodium selenite, 170 mg alpha-tocopherol or a placebo for 4 wk. Activities of glutathione-S-transferase, superoxide dismutase and catalase were assayed in erythrocytes, plasma and platelets and in liver and muscle biopsy tissues. No changes in activities of any of these tissue enzymes were observed in any of the three groups. There were also no changes in non-selenium dependent glutathione peroxidase activities in liver or plasma. The lack of changes in any of these enzymes following selenium supplementation suggests that adaptive changes to the low selenium status of these subjects had not occurred through these lipid peroxidation defense mechanisms.

Adult↗

Capillary whole blood selenium determination in assessing selenium status of children.

Selenium levels in venous and capillary whole blood of children were assessed by direct graphite furnace atomic absorption spectrometry. Selenium data obtained for both blood samples did not differ statistically (p > 0.05). Capillary blood may therefore be used as an alternative to venous blood, which is more difficult to obtain from children. Whole-blood selenium data for children in the Antwerp region (Belgium) were compared with literature data for children from other countries.

Belgium↗

Selenoimidoylation of alcohols with selenium and isocyanides and its application to the synthesis of selenium-containing heterocycles.

The reaction of alcohols with selenium and isocyanides in the presence of DBU gave oxyimidoylselenoates 6. Trapping of 6 with BuI resulted in high-yield formation of selenocarbonimidates 4. When alk-2-yn-1-ols 9 were allowed to react with selenium and isocyanides under similar conditions, new selenium-containing heterocycles 10, 2-imino-4-alkylidene-1,3-oxaselenolanes, were obtained via cycloaddition of oxyimidoylselenoates 13 generated in situ by intramolecular addition of selenolates to carbon-carbon triple bonds.

Journal Article↗

Selenium (Se) from high-selenium broccoli is utilized differently than selenite, selenate and selenomethionine, but is more effective in inhibiting colon carcinogenesis.

The reduction in incidence of chemically-induced colon cancer by foods high in selenium (Se) was investigated in Fisher-344 rats. The foods used were high-Se broccoli (produced in a greenhouse by addition of selenate to the media surrounding the plant roots) and a processed high-Se wheat product (made by milling high-Se wheat purchased from a seleniferous area). Weanling rats were fed diets containing different amounts of Se from these foods or from selenium salts (selenite and selenate). Early in the experiment the animals were injected with a chemical carcinogen. After 11 weeks on diets animals were killed and the colons examined for preneoplastic lesions (aberrant crypts foci, ACF). ACF were significantly reduced in animals fed supra-nutritional amounts of Se from broccoli, despite the finding that Se from broccoli was poorly bioavailable. Supra-nutritional amounts of Se from high-Se processed wheat also significantly reduced aberrant crypts (AC), although pure selenomethionine, (the predominant chemical form of Se in wheat), did not significantly reduce AC. These results emphasize the need to study Se in food forms, and not extrapolate from previous studies using pure chemical forms in cancer inhibition studies. They also demonstrate that foods with high Se bioavailability are not necessarily the most efficacious for cancer incidence reduction.

Animals↗

Stability of total selenium and selenium species in lyophilised oysters and in their enzymatic extracts.

To obtain reliable information on speciation analysis it is necessary to previously evaluate the stability of the species in the sample of interest. Furthermore, in those cases in which sample treatment to extract the species is time-consuming, an evaluation of how to maintain species integrity in the extracts is paramount. Thus, the present paper reports the stability of total Se, SeMet and TMSe+ in freeze-dried oyster and in the enzymatic extracts stored in Pyrex and polyethylene containers at different temperatures (-18, 4 and 20 degrees C). Total selenium determinations and Se speciation were carried out by HG-AAS after acid digestion in a microwave oven and by on-line coupling of cation exchange HPLC-ICP-MS after enzymatic hydrolysis, respectively. The results obtained for the freeze-dried sample showed that total Se and the selenium species evaluated are stable for at least 12 months, under all the conditions tested. However, Se species in the enzymatic extracts are only stable for 10 days if stored at 4 degrees C in Pyrex containers. These results show that the extracts do not necessarily have to be analysed just after sample treatment.

Animals↗

Kale carotenoids are unaffected by, whereas biomass production, elemental concentrations, and selenium accumulation respond to, changes in selenium fertility.

Selenium (Se) is a micronutrient in mammalian nutrition and is accumulated in kale (Brassica oleracea L. var. acephala), which has high levels of lutein and beta-carotene. Selenium, lutein, and beta-carotene have important human health benefits and possess strong antioxidant properties. The objectives of this study were to determine the influence of different Se [as sodium selenate (Na(2)SeO(4)) and sodium selenite (Na(2)SeO(3))] fertility levels on (1) biomass accumulation, (2) the accumulation patterns of carotenoid pigments, and (3) elemental accumulation in the leaves of kale. Winterbor kale was greenhouse-grown using nutrient solution culture with Se treatment concentrations of 0.0, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, and 3.5 mg Se/L as Na(2)SeO(4) and 0.0, 0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 mg Se/L as Na(2)SeO(3). Increases in either selenate (SeO(4)(-)(2)) or selenite (SeO(3)(-)(2)) resulted in decreases in kale leaf tissue biomass. Neither of the Se treatments had an effect on the accumulation of lutein or beta-carotene in leaf tissues. Increasing SeO(4)(-)(2) significantly increased the accumulation of kale leaf Se; however, leaf tissue Se did not significantly change over the SeO(3)(-)(2) treatments. Increases in SeO(4)(-)(2) affected the leaf tissue concentrations of P, K, Ca, Mg, S, B, Cu, Mn, and Mo, whereas SeO(3)(-)(2) only affected B and S. Growing kale in the presence of SeO(4)(-)(2) would result in the accumulation of high levels of tissue Se without affecting carotenoid concentrations.

Brassica↗

Effect of selenium source on selenium status, neutrophil function, and response to intramammary endotoxin challenge of dairy cows.

The effects of feeding dry and early lactation dairy cows diets with selenate or selenized yeast (Se-yeast) on concentrations of Se in serum, milk, and newborn calves, neutrophil function, and inflammatory response were determined. At 60 d before anticipated calving until approximately 30 d in milk (DIM), cows were fed diets that contained 0.3 mg of supplemental Se/kg of DM from sodium selenate or Se-yeast. Diets also contained 0.2% supplemental S (as sulfate) because it has been shown to reduce absorption of Se by dairy cows. The concentration of Se in serum at calving and 28 DIM was about 1.4 times greater for cows fed Se-yeast than for those fed selenate. Serum concentrations decreased 45 and 23% from dry-off to calving for cows fed selenate or Se-yeast, respectively. Selenium concentrations in serum from newborn calves were also about 1.4 times greater when the dams were fed Se-yeast. Concentrations of Se in colostrum and milk were about 1.8 times greater when cows were fed Se-yeast. Blood neutrophils were isolated from cows at 28 DIM and were used in an in vitro kill assay. Selenium treatment did not affect bacterial kill or the percentage of neutrophils that phagocytized bacteria. At approximately 28 DIM, one quarter from each cow was infused with a solution containing endotoxin. Peak body temperature (40.7 degrees C) occurred 6 h postinfusion, and peak somatic cell count (6.5 log10/mL) occurred at 12 h postinfusion. Neither measure was influenced by Se treatment.

Animals↗