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Studies on vitamin E and selenium deficiency in young pigs. II. The hydrogen peroxide hemolysis test and the measure of red cell lipid peroxides as indices of vitamin E and selenium status.

The usefulness of the hydrogen peroxide hemolysis test and the measure of red cell lipid peroxides as indices of vitamin E and selenium deficiency in swine has been evaluated. Results indicated that although the hydrogen peroxide hemolysis test may be of some indication of the vitamin E status, it is not a reliable index of vitamin E deficiency in swine, at least on an individual basis. In contrast, the measure of red cell lipid peroxides can be considered a reliable test for vitamin E deficiency in swine. The hydrogen peroxide hemolysis test and the red cell lipid peroxides were not significantly affected by selenium deficiency.

Animals↗

Selenium regulation of selenium-dependent glutathione peroxidases in animals and transfected CHO cells.

Glutathione peroxidase (GPX1) was the first identified selenium-dependent enzyme, and this enzyme has been most useful as a biochemical indicator of selenium (Se) status and the parameter of choice for determining Se requirements. We have continued to study Se regulation of GPX1 to better understand the underlying mechanism and to gain insight into how cells themselves regulate nutrient status. In progressive Se deficiency in rats, GPX1 activity, protein and mRNA all decrease in a dramatic, coordinated and exponential fashion such that Se-deficient GPX1 mRNA levels are 6-15% of Se-adequate levels. mRNA levels for other Se-dependent proteins are far less decreased in the same animals. The mRNA levels for a second Se-dependent peroxidase, phospholipid hydroperoxide glutathione peroxidase (GPX4), are little affected by Se deficiency, demonstrating that Se regulation of GPX1 is unique. Se regulation of GPX1 activity in growing male and female rats shows that the Se requirement is 100 ng/g diet, based on liver GPX1 activity; use of GPX1 mRNA as the parameter indicates that the Se requirement is nearer to 50 ng Se/g diet in both male and female rats. This approach will readily detect an altered dietary Se requirement, as shown by the incremental increases in dietary Se requirement by 150, 100 or 50 ng Se/g diet in Se-deficient rat pups repleted with Se for 3, 7 or 14 d, respectively. Studies with CHO cells stably transfected with recombinant GPX1 also show that overexpression of GPX1 does not alter the minimum level of media Se necessary for Se-adequate levels of GPX1 activity or mRNA. We hypothesize that classical GPX1 has an integral biological role in the mechanism used by cells to regulate Se status, making GPX1 an especially useful and effective parameter for determining Se requirements in animals.

Animals↗

Condensed thiophenes and selenophenes: thionyl chloride and selenium oxychloride as sulfur and selenium transfer reagents.

3,4-Cyanomethyl substituted thiophenes reacted with thionyl chloride in the presence of base to give dicyano substituted thieno(3,4-c)thiophenes. The use of selenium oxychloride furnished the corresponding cyano substituted seleno(3,4-c)thiophene. 1,2-Phenylenediacetonitriles gave the corresponding cyano substituted benzo(c)thiophenes and benzo(c)selenenophenes, respectively, upon reaction with thionyl chloride and selenium oxychloride in the presence of base.

Journal Article↗

The effect of long-acting injectable selenium formulations on blood and liver selenium concentrations and liveweights of red deer (Cervus elaphus).

AIM: To evaluate the efficacy of a new long acting injectable selenium ( Se ) formulation to increase the Se status and prevent Se deficiency in red deer. METHODS: Groups of weaned red deer (four stags and six hinds/group) grazing pastures containing <30 mg Se/kg DM were injected subcutaneously with either 0.5, 1.0 or 2.0 mg Se/kg as a new formulation of BaSeO4 (Deposel Multidose), 1.0mgSe/kg of a current formulation (Deposel), or not treated. Blood Se concentrations and liveweight were measured nine times at intervals over 377 and 270 days, respectively. RESULTS: Both formulations of Se elevated blood Se concentrations from 105 nmol/l pre-injection for at least 377 days with peak levels of 1894, 1395 and 818 nmol/l for high, medium and low doses of Deposel Multidose, respectively, at 73141 days, and 1508 nmol/l at 73-141 days for the medium dose of Deposel, which persisted at similar levels for the duration of the study. Deposel Multidose produced fewer and less severe subcutaneous tissue reactions than Deposel. Pastures contained 10 to 30 mg Se/kg DM. There was no significant difference in growth rate between treated and control deer. There was a significant (p<0. 01) linear relationship (y = 1.25x + 71.6, R2=0.86) between blood (x) and liver (y) Se concentrations in the range of 120 - 2100 nmol/l for blood concentrations, and 200 - 3000 nmol/kg for liver concentrations. CONCLUSION: Injections of BaSeO4 in both formulations studied were effective in increasing the Se status of deer but the new formulation produced fewer and less-severe tissue reactions. Young growing red deer appear less sensitive to Se deficiency as measured by weight gain, than sheep and cattle, suggesting that reference ranges for those species are not appropriate for deer. There was a linear correlation between blood and liver selenium concentrations.

Journal Article↗

[Effects of selenium and germanium on lipid peroxidation in rats fed with low-selenium grain].

Rats were fed with grain produced in the area prevalent of Keshan disease for 12 weeks, by adding certain amount of sodium selenite (Na2SeO3) and carboxylethylgermanium (Ge-132) to study the combined effects of selenium and germanium on lipid peroxidation and metabolism of free radicals in their bodies. Results showed selenium and germanium added to the grain could reduce the content of free radicals in rats' liver and kidney tissues, and that of lipid peroxide in their heart, liver and kidney, and increase the activities of glutathione peroxidase (GSH-Px) in their blood, in a synergic way.

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↗

The effect of barium selenate injection on selenium concentration and glutathione peroxidase activity in blood of pregnant ewes fed selenium-deficient diet.

Selenium (Se) levels in whole blood and plasma, and glutathione peroxidase (GSH-Px) activities in red cells and plasma were measured in ewes fed an Se-deficient diet injected with barium selenate before breeding season. Highly significant increases in Se levels and GSH-Px activities (P less than 0.001) were observed throughout the gestation period and during lactation. In the control group, Se levels and GSH-Px activities decreased significantly (P less than 0.001), and were at critically low levels during lambing and lactation periods.

Animals↗

Human selenium supplementation as assessed by changes in blood selenium concentration and glutathione peroxidase activity.

This review concentrates on the differences in the effects on indicators of selenium (Se) status of different chemical forms proposed for Se supplementation in healthy subjects. Modifications in blood (plasma and erythrocyte) Se concentrations and in the Se-dependent glutathione peroxidase (GSH-Px) enzymatic activity, in plasma, erythrocytes and platelets, were compared considering the basal Se status, the dose and the chemical form of Se supplement, as well as the duration of the intervention. Organic Se forms (Se-yeast, selenomethionine and food-Se) increased blood Se more concentration rapidly and to a greater extent than inorganic forms (selenite and selenate). However, no significant difference in the response of both plasma and erythrocyte GSH-Px activity could be observed. In contrast, platelet GSH-Px activity was more sensitive to the chemical form of Se administered. Saturation of platelet GSH-Px activity occurred at lower plasma Se levels when selenite or selenate were used as Se supplements than with the organic forms. Interpretation of some of the results is provided and ideas are suggested for consideration in further studies.

Blood Platelets↗

Composition of chemical species of selenium contained in selenium-enriched shiitake mushroom and vegetables determined by high performance liquid chromatography with inductively coupled plasma mass spectrometry.

Selenium (Se) species in Se-enriched shiitake mushroom (Lentinula edodes) were identified and quantified by high performance liquid chromatography with inductively coupled plasma mass spectrometry (HPLC-ICPMS). Two types of Se-enriched shiitake obtained from selenite- or selenate-fertilized mushroom beds were used. More than 80% of Se in both shiitake samples could not be extracted with 0.2 M HCl. Protease digestion released a large amount of selenomethionine from the shiitake enriched with selenite. However, most of the Se in the shiitake enriched with selenate was not released by protease but was released by a cell wall digestive enzyme and most of the Se released was identified as selenate. These results indicate that the main Se species in the shiitake enriched with selenite or selenate is selenomethionine bound to protein or selenate bound to polysaccharides in the cell wall, respectively. Several Se-enriched vegetables grown on a soil fertilized with selenate were also analyzed by HPLC-ICPMS. Four Se species, selenate, Se-methylselenocysteine, selenomethionine, gamma-glutamyl-Se-methylselenocysteine, and an unknown Se compound were detected in the vegetables. The composition of Se species varied with the kinds or parts of vegetables. The main Se species in bulbs, leaves or flowers of the Se-enriched garlic, onions, cabbage and ashitaba were selenate, Se-methylselenocysteine or gamma-glutamyl-Se-methylselenocysteine, while those in fruit bodies of the peppers and pumpkin were selenomethionine bound to protein. Bioavailabilities of Se in the shiitake mushroom enriched with selenite and the vegetables enriched with selenate are expected to be high, but that in shiitake enriched with selenate may be low.

Chromatography, High Pressure Liquid↗

Effect of intermittent supplementation with selenate on selenium status of rats fed selenium-deficient diet.

To examine the selenium (Se) status of rats intermittently supplemented with Se, we measured tissue Se contents and glutathione peroxidase (GPx) activities in rats fed a Se-deficient diet intermittently supplemented with selenate. In experiment 1, four groups of male 4-wk-old Wistar rats were fed a Torula yeast-based Se-deficient diet (Se content, < 0.01 microg/g) for 28 d. During the experimental period, the diet of each group was supplemented with sodium selenate (0.17 microg Se/g) for 0, 1, 2 or 7 d/wk. The tissue Se contents and GPx activities both increased gradually with an increase in frequency of the selenate supplementation, and significant linear regressions were observed between the frequency and these Se indices. In particular, the correlation coefficient in the liver and plasma indices was nearly equal to a value of 1.0. In experiment 2, three groups of rats were fed the Se-deficient basal diet for 28 d. Among these, one group was daily supplemented with sodium selenate to the Se-deficient diet at a level of 0.17 microg Se/g, and another group was intermittently supplemented with the selenate at a level of 1.19 microg Se/g for 1 d/wk. The tissue Se contents and GPx activities both were increased by the selenate supplementation and no significant difference was observed between daily and weekly supplementation in the Se indices except in erythrocyte Se. These results indicate that Se status in the growth period is dependent on total Se intake in this period and that weekly intermittent supplementation with Se can maintain adequate Se status.

Animals↗

Inhibitory effects of selenium on 1,2-dimethylhydrazine and methylazoxymethanol colon carcinogenesis: correlative studies on selenium effects on the mutagenicity and sister chromatid exchange rates of selected carcinogens.

Selenium (Se) inhibition of either the activation of test compounds and/or mutagenic events elicited by activated compounds is suggested by experimental rat assays, mutagenesis assays, and assays with human lymphocytes in culture. The colon tumor incidence in 1,2-dimethylhydrazine (DMH)-treated rats was reduced from 87% to 40% by 4 ppm Se supplements in the drinking water. Supplemental Se decreased the total number of colon tumors induced by DMH more than three-fold and by methylazoxymethanol (MAM) almost two-fold. Coexposure of Salmonella typhimurium TA 1538 to an effective molar ratio of Se/2-acetylaminofluorene=10, Se/N-OH-acetylaminofluorene=10 and SE/N-OH-aminofluorene=300 reduced the mutagenicity to 65, 68, and 61% of their respective controls with mutagen alone. With a molar ratio of Se/N-OH-AAF=100, Se reduced the activity to 28% of the mutagenicity of N-OH-AAF alone. Preliminary data indicating MAM is mutagenic in S. typhimurium TA 1535 and His G 46(6837) are presented. In toxicity studies exposure of human lymphocyte cultures to 1.3 X 10(-9) to 1.6 X 10(-5) M Se yielded sister chromatid exchange (SCE) rates equivalent to background levels of 6--7 SCE per cell. The SCE frequencies of lymphocytes cultured with Se and selected carcinogens are discussed.

Animals↗

Effects of elevated selenium concentration on selenium accumulation and nitrogen fixation symbiotic activity of Melilotus indica L.

Biological and soil factors which contribute to the successful colonization of an annual legume species. Melilotus indica L., in soils with elevated selenium (Se) were studied. This species was introduced into the Kesterson Reservoir in the fresh top soil that was brought in under the Kesterson Cleanup Action to fill lowering pond sites and prevent the formation of ephemeral pools containing hazardous levels of Se. In 4 years since its introduction, it has expanded its range of colonization from the fresh soil fill sites to the native soil sites and contributed 10 to 50% of biomass to the grassland communities. The plant and nodule tissue Se concentrations of the field grown plants were found to be negatively correlated with the soil sulfate concentration. Nutrient solution culture studies discovered that M. indica was able to accumulate 500 micrograms Se g-1 dry weight without a reduction of growth rate. Plants without nodulation were found to accumulate a greater amount of Se and more vulnerable to Se toxicity. Acetylene reduction rate measurements indicate that the nitrogen fixation symbiotic activity appears to be more susceptible to an elevated Se concentration than its host plant. M. indica is a winter weed, and it occurs naturally in the Se-rich soils. It grows actively over the winter and spring and complete its life cycle in May. If the root nodules and root tissues are incorporated into the soil, the rate of soil Se volatilization may be accelerated over the warm summer months. For disposal of the Se-rich plant materials the plant shoot tissues may be harvested for Se-deficient forage supplementation. Therefore, this species may be useful for field management and reclamation of Se-contaminated soils.

Biomass↗

Bioextraction of selenium by forage and selected field legume species in selenium-laden soils under minimal field management conditions.

A forage plant, tall fescue (Festuca arundinacea), and a selected field legume species, sour clover (Melilotus indica), were examined for their selenium (Se) bioextraction abilities in Se-laden soils under minimal management conditions. Natural vegetations in a 2-acre plot adjacent to the forage plots were also studied for Se accumulation comparisons. During the dry season, in the fall of 1994, the field plots were either irrigated weekly or without irrigation. No fertilization and weed control were applied. The plants were harvested in May 1995. There were considerable differences in the ability of Se uptake between the forage and the legume species and among the naturally established plant species; the amount of Se accumulated per land area was largely dependent on their respective biomass production. Comparing Se concentration between preplant and postharvest, there was a detectable reduction in the soil selenate, selenite, and water-extractable organic Se in the tall fescue and melilotus plots. The field irrigation provided more favorable conditions for bioextractions and dissipation of Se by the plants. However, the available soil Se only accounts for less than 10% of the total soil Se and no detectable reduction of total soil Se was found. This may be due to the large inventory and variation of Se concentrations in the field soils and therefore obscured the detectable differences. For practical considerations, the forage plants can be repeatedly harvested and used for rangelands of Se deficiency currently seen in some northern California counties.

Analysis of Variance↗

Absorption and distribution of selenium in animals consuming canola grown for selenium phytoremediation.

Canola (Brassica napus) grown as a selected plant species for field phytoremediation of selenium (Se) may be harvested and utilized as Se-enriched forage for marginally Se-deficient lambs and cows. Two field studies were conducted under controlled conditions to evaluate the accumulation of Se into different animal tissues, including blood, excreta, and milk. In Study 1, treatments consisted of feeding lambs freshly cut Se-enriched canola (containing approximately 4 mg Se kg(-1) DM) or control canola (containing <0.1 mg Se kg(-1) DM), respectively, for 64 days. In Study 2, treatments consisted of feeding cows dried Se-enriched canola (containing approximately 3.5 mg Se kg(-1) DM) as part of their daily ration for 20 days. In Study 1 at postmortem, Se concentrations were significantly greater in all tested tissues and in excreta from lambs fed Se-enriched canola. In Study 2, Se values were slightly higher in blood and excreta, but not significantly higher in milk from cows sampled throughout the study. Significant differences in total live animal weight were not observed between treatments in either study. Based on these results, canola plants (not including seeds) used for field phytoremediation of Se may be harvested and safely fed to lambs and cows to help meet normal Se intake requirements.

Absorption↗

Effect of selenium supplementation on the distribution of selenium among plasma proteins of a patient with maple syrup urine disease.

Selenium (Se) status was studied in a patient with classical maple syrup urine disease (MSUD) receiving Se supplement. The basal plasma Se concentration was 0.06 mumol/l increasing to 2.1 mumol/l after 40 days of supplementation. When the plasma Se distribution was analysed by gel filtration, a major peak was seen close to the high molecular weight proteins with a second peak in the albumin region. When the Se dose was decreased in a stepwise manner from 50 micrograms/day to 25 micrograms/day and then to 17 micrograms/day plasma Se decreased, but the proportion of plasma Se in the two protein peaks did not change. In a healthy girl not supplemented with Se, the proportion of plasma Se in the albumin region was somewhat lower. In the MSUD patient glutathione peroxidase activity was initially low, and increased ten-fold during Se supplementation. The study indicates that the Se requirement for plasma glutathione peroxidase activity was fulfilled at the lowest dose of Se used and that Se is incorporated into several plasma proteins after supplementation.

Blood Proteins↗

[Hematological parameters, selenium concentration and glutathione peroxidase activities in serum and the liver of rats at different selenium and vitamin E levels].

The aim of the both experiments was to determine whether selenium or selenium/vitamin E supply of rats significantly influences the most important hematological criteria. With experiment 1 the influence of Se deficiency should be determined at two different times of growing. So 36 weaned rats were divided into 2 groups of 18 animals each, the half of them being decapitated at day 22, the rest on day 45. In experiment 2 with the aim to investigate a combination of deficient, adequate and excessive Se and vitamin E supply 90 weaned rats in 9 groups were decapitated at day 44. The basic diet contained 0.04 mg Se and 8 mg vitamin E per kg dry matter and was supplemented in exp. 1 with 0 mg or 0.2 mg Se and 30 mg vitamin E and in exp. 2 with 0 mg, 0.2 mg or 1.0 mg Se and 0 mg, 30 mg or 200 mg vitamin E. With Se deficiency Se concentration and GSH-Px activity in serum and liver were significantly reduced. With excessive Se supply Se concentration in serum was higher; there was no effect on GSH-Px activity. Vitamin E supply had no influence neither on Se content nor on GSH-Px activity in serum or in liver. In exp. 1 Se deficiency caused no clear changes of the analysed hematological criteria although the increase of MCV (+3%) and hematocrit (+7%) on day 22 and the increase of leucocytes (+43%) and the decrease of MCH (-3%) and MCHC (-6%) on day 45 were statistically significant. In exp. 2 these results could not be repeated. The vitamin E supply was without significant effects on the examined hematological parameters.

Animals↗

Effect of selenium and protein deficiency on selenium and glutathione peroxidase in rats.

Twenty-four weanling male Wistar rats were divided into four groups fed diets containing adequate or deficient levels of selenium (0.5 ppm [+Se] or < 0.02 ppm [-Se] and protein (15% [+Pro] or 5% [-Pro]), but adequate levels of all other nutrients for 4 wk to determine the effects of Se deficiency and protein deficiency on tissue Se and glutathione peroxidase (GSHPx) activity in rats. Plasma, heart, liver, and kidney Se and GSHPx were significantly lower in Se-deficient groups in relation to Se-sufficient groups. In Se-deficient groups, Se and GSHPx were significantly higher in -Se-Pro rats in heart, liver, and kidney. Data analysis showed that there were significant interaction effects between dietary Se and protein on Se and GSHPx of rats. It is assumed that under the condition of Se deficiency, a low level of protein may decrease Se and GSHPx utilization, increase GSHPx synthesis, and result in Se redistribution. This could account for high levels of Se and GSHPx in the -Se-Pro rats compared to -Se+Pro rats.

Analysis of Variance↗