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The effect of neem (Azadirachta indica) extract and dietary selenium on distribution of selenium in hepatocarcinogenesis induced rat.

Neem, Azadirachta indica, is a plant from the family Meliaceae, known as "Pokok Semambu" in Malay community. It has been extensively used in India as traditional Ayurvedic and folklore minedicine for the treatment of various diseases. This study aimed to determine the distribution of selenium in the liver of rats during hepatocarcinogenesis when neem aqueous extract and dietary selenium was supplemented.

Administration, Oral↗

Selenium concentration and glutathione peroxidase activity in selenium and magnesium deficient rats.

To clarify the effects of selenium (Se) and magnesium (Mg) deficiencies on Se and glutathione peroxidase (GSHPx) status, weanling male Wistar rats weighing 50-60 g were placed on four kinds of diets divided by two levels of Se (0.5 or 0.019 mg/kg) and Mg (500 or 50 mg/kg) for 8 wk. Magnesium deficiency had an influence on distribution of Se, which was increased in muscle and decreased in other tissues. The changes in GSHPx matched those in Se. The levels of Se and GSHPx in most tissues were lower in Se-Mg-deficient rats than in Se-deficient rats. Thus, selenium and Mg deficiencies would make oxidant lesion more serious than Se deficiency.

Animals↗

Improving selenium extraction by sequential enzymatic processes for Se-speciation of selenium-enriched Agaricus bisporus.

Sample preparation methods based on the use of proteolytic and cell wall digesting enzymes for the speciation analysis of selenized mushroom were investigated. The sample (Agaricus bisporus; 160 microg total Se per g sample) was grown on compost supplemented with selenized yeast. Experiments were carried out to elucidate the possible role of the cell wall digesting enzymes--Lysing enzyme and Driselase--in the improvement of extraction efficiency with and without inhibiting proteolysis during cell wall digestion. A 3-step procedure applying Lysing enzyme and pronase gave the highest extraction efficiency (89%); however, the best species recovery was achieved by a one-step proteolytic procedure. All the procedures of selenium speciation were controlled by independent ICP-AES analysis measuring the total amount of selenium.

Agaricus↗

Measurement of total selenium and selenium(IV) in seawater by stripping chronopotentiometry.

We developed a stripping chronopotentiometric method (constant current stripping analysis, CCSA) with a mercury film electrode for selenium quantification in seawater. A sensitivity and detection limit of 222 ms ng(-1) l and 4 ng l(-1) (50 pM), respectively, were accomplished for a 3-min electrolysis time. Compared to the other chronopotentiometric methods available for a single selenium measurement only in natural waters, our procedure exhibits a ten times better sensitivity. It, therefore, allows one to reach the current concentration thresholds found in coastal and oceanic waters (30-200 ng l(-1)). Moreover, a simple change in operating conditions enables one to also quantify Se(IV), a toxic dissolved species. With respect to the other electrochemical methods of current use, our procedure is beneficial because of its ease-of-use: it needs neither degassing step, nor catalyser.

Chlorides↗

Active transcription of the selenium-dependent glutathione peroxidase gene in selenium-deficient rats.

Selenium-dependent glutathione peroxidase (Se-GSH-Px, Ec.1.11.1.9) is the best characterized selenoenzyme in higher animals. However, neither the mechanism whereby selenium (Se) becomes incorporated into the enzyme nor the level at which the expression of Se-GSH-Px gene is regulated by Se is fully understood. In the current investigation, we have determined the relative rates of the transcription of the Se-GSH-Px gene in purified liver nuclei isolated from rats fed on Se-supplemented or Se-deficient diets. No significant difference in the transcription rates appeared in these two groups. These results are consistent with the previous observations that active message for Se-GSH-Px- that is, translatable mRNA for Se-GSH-Px- is present in Se-deficient tissues (Li et al., J. Biol. Chem., 265, 108-113, 1990). The data also suggest that the alteration of Se-GSH-Px activity and the corresponding protein and mRNA levels in rats subjected to dietary Se manipulation can be attributed only to post-transcriptional regulation.

Animals↗

Selenium as a sulfhydryl redox catalyst and survey of potential selenium-dependent enzymes.

The ability of selenium (Se) to act as a redox catalyst is an important factor in understanding the biological function of selenoproteins in addition to that of GSH peroxidase. Selenocystine at micromolar levels exhibited pseudothiotransferase activity by enhancing the reduction of 5,5-dithiobis-(2-nitrobenzoic acid) (DTNB) by thiols. In contrast, selenite inhibited the reduction of DTNB by thiols. Selenite was more catalytic than selenocystine in the reduction of cytochrome c by GSH, whereas GSH peroxidase was a weak catalyst. Tissues from Se-deficient and Se-supplemented rats were assayed for activities of GSH-thiotransferase, NADPH cytochrome c reductase, formaldehyde dehydrogenase, and a hypothesized GSH cytochrome c reductase. GSH-thiotransferase activity was significantly increased in the liver of Se-deficient rats. No appreciable activity of this enzyme was found in the kidney of rats from either dietary group. No enzymatic activity for cytochrome c reduction by GSH was detected in cytosols, mitochondria, or microsomes from liver and kidney of Se-deficient or Se-supplemented rats. Formaldehyde dehydrogenase was significantly higher in liver cytosols from Se-supplemented rats than from Se-deficient rats. The higher activity was not attributed to Se-containing proteins, but to an unknown small molecular-weight factor. This study did not support the hypothesis that physiological levels of Se may be involved in sulfhydryl-disulfide exchange reactions in vivo, or that selenium may enhance cytochrome c reduction by GSH in vivo.

Animals↗

Purification and characterization of selenium-containing phycocyanin from selenium-enriched Spirulina platensis.

A fast protein liquid chromatographic method for purification of selenium-containing phycocyanin (Se-PC) from selenium-enriched Spirulina platensis was described in this study. The purification procedures involved fractionation by ammonium sulfate precipitation, DEAE-Sepharose ion-exchange chromatography and Sephacry S-300 size exclusion chromatography. The purity ratio (A620/A280) and the separation factor (A620/A655) of the purified Se-PC were 5.12 and 7.92, respectively. The Se concentration of purified Se-PC was 496.5 microg g(-1) protein, as determined by ICP-AES analysis. The purity of the Se-PC was further characterized by UV-VIS and fluorescence spectrometry, SDS-PAGE, RP-HPLC and gel filtration HPLC. The apparent molecular mass of the native Se-PC determined by gel filtration HPLC was 109 kDa, indicating that the protein existed as a trimer. SDS-PAGE of the purified Se-PC yielded two major bands corresponding to the alpha and beta subunits. A better separation of these two subunits was obtained by RP-HPLC. Identification of the alpha and beta subunits separated by SDS-PAGE and RP-HPLC was achieved by peptide mass fingerprinting (PMF) using MALDI-TOF-TOF mass spectrometry.

Chromatography, Liquid↗

Selenium protection against mercury toxicity: high binding affinity of methylmercury by selenium-containing ligands in comparison with sulfur-containing ligands.

In determining the protection of selenium against mercury toxicity, the binding affinity of methylmercury by various selenium-containing ligands was investigated by proton magnetic resonance (PMR) spectroscopy. The most striking feature was the small J199Hg-1H value of the selenocysteamine- and selenocysteine-methylmercury complexes, namely, the high affinity of the selenohydryl group to the mercury in comparison with those of the sulfhydryl and amino groups. The order of binding affinity of the coordination groups toward methylmercury is clearly SeH greater than SH greater than or equal to Se-Se greater than NH2 greater than S-S, SeCH3, SCH3. A definite correlation was found to exist between the mercury-proton coupling constants and the chemical shifts of methyl groups of the methylmercury complexes. A relationship between the order (Se greater than S greater than NH2) of affinity for methylmercury and the basicity (or electronegativity and covalent radius) of the donor groups was also discussed. These results suggest the high covalency of the CH3Hg-Se bond, which involves dpi-dpi back bonding.

Chemical Phenomena↗

Characterization of the regional distribution of selenium in Chile using selenium in hens' eggs as a monitor.

Regional distribution of selenium (Se) in Chile was investigated by using the concentration of Se in hens' eggs as a monitor. Forty-one locations along the entire length of the country were sampled. Average (+/- SD) egg-white Se content (mg/kg dry) was 0.79 +/- 0.41, range 0.22-2.23. Corresponding yolk Se values were, mean 0.81 +/- 0.43, (mg/kg dry) range 0.26-2.23. Locations grouped in five main areas, according to their geographic-climatic characteristics, showed significant differences regarding both egg-white Se and yolk Se. Analyzed dietary Se intake from two distinct areas reflected the trends observed in the Se content of egg fractions from such regions. These data support the utilization of the concentration of Se in hens' eggs as a useful monitor of dietary selenium consumed by selected populations.

Animals↗

Selenium removal by constructed wetlands: quantitative importance of biological volatilization in the treatment of selenium-laden agricultural drainage water.

Management of selenium (Se) -contaminated agricultural drainage water is one of the most important environmental issues in California. To evaluate the feasibility of utilizing constructed wetlands to remediate Se-laden drainage water and the role of biological volatilization in Se removal, 10 flow-through wetland cells were constructed in 1996 in Corcoran, California. The monthly monitoring study from May 1997 to December 1999 showed that the vegetated wetlands were capable of significantly reducing Se from the inflow drainage water; an average of 69.2% of the total Se mass in the inflow was removed. Most of the Se was retained in sediment, and <5% of the Se was accumulated in plant tissues. Selenium volatilization was highest in the rabbitfoot grass wetland cell, where 9.4% of the Se input was volatilized over a 2-year period. Volatilization was greater in spring and summer than in fall and winter. For example, in May and June of 1998, 35 and 48%, respectively, of the Se entering the rabbitfoot grass cell was volatilized, whereas in the winter months, <5% was volatilized. The feasibility of using constructed wetlands for Se remediation, methods for the enhancement of Se volatilization, and the importance of considering potential Se ecotoxicity are discussed.

Agriculture↗

Selenium accumulation in beef: effect of dietary selenium and geographical area of animal origin.

Selenium (Se) is an essential nutrient with multiple human health benefits; the single most important dietary source of Se is beef. The Se content of beef varies, and cattle fed a high selenium diet may have Se concentrations in beef that are well above average. Such beef is potentially a unique supplemental source of dietary Se. To examine factors affecting Se accumulation in beef, 16 steers (initial wt 374.4 +/- 33.7 kg) were taken from seleniferous or nonseleniferous areas and fed in a 2 x 2 factorial design with diets high or moderate in Se (11.9 or 0.62 mg Se/kg diet). Diets contained 50% alfalfa, 25% wheat, and 25% corn on a dry matter basis. All dietary Se was from agricultural products, and Se in the high Se diet was primarily from high Se wheat and alfalfa hay. A loin muscle biopsy was taken at the start of the trial to determine initial Se content of beef. Steers were slaughtered after 14 weeks of the trial, and edible carcass (round, sirloin, shoulder clod, and ribeye) and organ samples were collected. Diets did not affect growth or feed intake (P > 0.05), and Se toxicity signs were not observed. Different cuts of meat had similar Se concentrations, and the Se content of all cuts was increased by both high dietary Se and high Se background. Except for liver and kidney, Se in tissues was increased by seleniferous background (P < 0.02) and high dietary Se (P < 0.001). Kidney Se concentrations of animals fed the high Se diet were lowest in animals from seleniferous areas (P = 0.04), suggesting a possible adaptation to the high Se diet. These results demonstrate that cattle fed diets high in Se from agricultural products will accumulate substantial amounts of Se in the beef without developing signs of Se toxicity and that prior Se status regulates Se accumulation in some organs. They further demonstrate that management practices may be altered so as to make beef a significant source of dietary Se.

Animals↗

Efficient selenium transfer from mother to offspring in selenoprotein-P-deficient mice enables dose-dependent rescue of phenotypes associated with selenium deficiency.

Mice deficient in selenoprotein P exhibit a disturbed selenium distribution and reduced activities of other selenoenzymes and display defects in growth and motor co-ordination. We have normalized selenoenzyme activities and rescued the phenotype of mutant mice by supplementing their nursing mothers with sodium selenite. Our results indicate that selenium from inorganic sources can be transferred efficiently via mother's milk to the developing offspring in a form that is both highly bioavailable by target tissues and yet sufficiently safe to prevent overdosages.

Administration, Oral↗

Selenium supplementation and selenium status of dairy cows fed diets based on grass, grass silage or maize silage.

In three separate trial series (TS) the effect of diet composition on selenium (Se) status of dairy cows were investigated. Diets were formulated based mainly on grass (TS1), grass silage (TS2) or maize silage (TS3) with different levels of Se supplementation. Each TS comprised a total of 30 dairy cows and contained one treatment group without Se supplementation (control) and two groups with increasing levels of Se supplementation (levels 1 and 2). Selenium was administered as Na-selenite. The control groups of the different TS showed a very low Se supply of 38-54 microg Se/kg DM. At level 1 the Se supply was increased to 102-165 microg Se/kg DM and at level 2 was 294-373 microg Se/kg DM. After completion of the 6-week trials the average plasma Se concentration of the control cows (without Se supplementation) across all TS was 21.5 microg/l; this increased significantly following Se supplementation, to 37.7 microg/l at level 1 and 61.5 microg/l at level 2. The plasma glutathione peroxidase (GSH-Px) activity of the control cows averaged 67 U/l, rising considerably after supplementation at level 1 to a value of 101 U/l, but showed little further increase at level 2 with a mean value of 120 U/l. By contrast, the average Se content of the milk was unchanged in the control and level 1 groups at 10.5 microg/kg and 10.9 microg/kg, respectively, and only increased markedly after supplementation at level 2 to a mean value of 15.1 microg/kg. The diet based on maize silage, while having a similar Se content as the grass and grass silage-based diets, resulted in a slightly improved Se status, which is due to a higher Se intake from soybean meal.

Animal Feed↗

Isolation of a selenium-containing thiolase from Clostridium kluyveri: identification of the selenium moiety as selenomethionine.

Clostridium kluyveri grown in the presence of 1 muM Na2(75)SeO3 produces a thiolase that copurifies with 75Se. Based on several criteria, the selenium moiety in this protein is selenomethionine. The 75Se-labeled amino acid in acid hydrolysates of the radioactive protein cochromatographed with authentic selenomethionine on an amino acid analyzer and on TLC plates in acidic and basic solvents. Incubation with S-adenosylmethionine synthetase and ATP converted the 75Se-labeled amino acid to a radioactive basic product that was indistinguishable from authentic Se-adenosylselenomethionine by ion exchange and TLC. The native selenoenzyme, Mr 155,000-158,000, is composed of four subunits of Mr 38,000-40,000. Thiolase of similar molecular weight that is less acidic and lacks selenium is also produced by C. kluyveri. The factors that control the relative levels of the two enzymes in the cell have not been identified.

Amino Acids↗

Bioavailability of selenium from raw and cooked ground beef assessed in selenium-deficient Fischer rats.

OBJECTIVE: The literature on the bioavailability of selenium (Se) from meats, especially beef, is meager, and that which existed when this research began suggested that Se was not highly bioavailable. In addition, much of the analytical values for Se in beef predated the Food and Drug Administration's 1973 approval of Se as an additive to feeds and mineral premixes of livestock. DESIGN: One hundred and thirty-six weanling female Fischer 344 rats were divided into two dietary groups: the selenium deficient group in which animals were fed a torula yeast (TY) basal diet which contained 0.008 mg/kg Se and the control group in which animals were fed the TY diet to which was added 0.10 mg/kg Se as sodium selenite. RESULTS: After 6 weeks of dietary treatment liver glutathione peroxidase (GSHPx) activity had fallen in the Se-deficient rats to 2.4% of that of control rats. At this time (week 6) rats from the Se-deficient TY diet were refed diets containing 0.10 mg/kg Se as selenite, selenate, raw or cooked ground beef that had been freeze-dried. During the Se-repletion period rats were sacrificed at weeks 1, 3, 5 and 8. Liver GSHPx activity and total Se levels in liver and muscle tissue were the criteria of Se bioavailability. After 8 weeks of Se resupplementation the recovery of liver GSHPx activity compared to the control animals (set at 100%) were selenite (98%, p > 0.05), selenate (117%, p < 0.05), raw beef (127%, p < 0.05) and cooked ground beef (139%, p < 0.05). Total Se in both liver and muscle tissue reflected the liver GSHPx activity with the total Se concentration in tissues being highest for cooked beef. CONCLUSION: The data suggest that bioavailability of Se from ground beef is greater than that from either selenite or selenate.

Animals↗

Selenium and cancer chemoprevention: hypotheses integrating the actions of selenoproteins and selenium metabolites in epithelial and non-epithelial target cells.

The trace element nutrient selenium (Se) discharges its well-known nutritional antioxidant activity through the Se-dependent glutathione peroxidases. It also regulates nuclear factor activities by redox mechanisms through the selenoprotein thioredoxin reductases. Converging data from epidemiological, ecological, and clinical studies have shown that Se can decrease the risk for some types of human cancers, especially those of the prostate, lung, and colon. Mechanistic studies have indicated that the methylselenol metabolite pool has many desirable attributes of chemoprevention, targeting both cancer cells and vascular endothelial cells, whereas the hydrogen selenide pool in excess of selenoprotein synthesis can lead to DNA single strand breaks, which may be mediated by some reactive oxygen species. We propose a new paradigm based on a consideration of the post-initiation biology of avascular early lesion expansion microenvironment, physiochemistry of Se delivery, and the obligatory need for angiogenesis to sustain lesion progression. Our model integrates the roles of selenoproteins and specific Se metabolites to account for cancer risk reduction or enhancement. For future studies, speciation (profiling) methods for Se metabolites and for Se forms in foods and supplements are much needed for hypothesis testing and for the development of mechanism-based Se status markers for cancer prevention. Randomized cancer prevention trials are necessary to test the efficacy of methyl selenium compounds.

Animals↗

Bioavailability of selenium to residents in a low-selenium area of China.

For 8 wk 5 groups of 10 men each were given 0.5 g/day DL-methionine, 150 micrograms Se/day as sodium selenite with or without methionine or 150 micrograms Se/day as selenomethionine with or without methionine. Twenty subjects received placebo as controls. Initially plasma Se rose more rapidly than RBC Se. Increases in Se levels were significantly greater with selenomethionine than with the selenite supplement. In the placebo and methionine supplemented groups neither plasma nor RBC Se varied significantly over the course of the study. Supplementation with selenium resulted in marked increases in plasma and RBC GSH-Px within 2 and 4 wk, respectively. Plasma and RBC GSH-Px activity did not differ significantly between Se-supplemented groups. These studies suggest that selenomethionine-Se was more effective in raising plasma and RBC Se than was selenite-Se. Methionine supplements may increase the bioavailability of selenium in severely deficient subjects.

Adult↗

Relationship of selenium-dependent glutathione peroxidase activity and nutritional pancreatic atrophy in selenium-deficient chicks.

Experiments were conducted to determine the relationship of selenium-dependent glutathione peroxidase (SeGSHpx) activity to the onset of nutritional pancreatic atrophy (NPA) in chicks. Chicks produced from hens fed a low Se, low vitamin E practical diet had low activities of SeGSHpx in plasma and pancreas at hatching. The SeGSHpx activity remained at a low level before and during all stages of the onset of NPA. Selenium supplementation of the diet prevented NPA and resulted in significant elevations in SeGSHpx activity. Although the early stages of NPA are believed to involve mitochondrial swelling, no significant differences were found in rate of oxygen uptake, respiratory control index or adenosine diphosphate to oxygen ratio between pancreatic mitochondria isolated from Se-deficient or Se-adequate chicks. Improvements in maternal Se status significantly increased chick pancreatic SeGSHpx activity at hatching and delayed the onset of NPA in chicks fed a Se-deficient diet. A significant proportion of the second-generation Se- and vitamin E-depleted chicks used in these studies was found to grow at nearly normal rates when fed the Se-deficient diet. These chicks, designated as refractory to the growth-depressing effect of severe Se-deficiency, were biochemically deficient (i.e., had very low SeGSHpx activities) and showed NPA. It is concluded that the Se-responsive lesion which results in NPA in the chick is different from that which results in depressed growth.

Animals↗