The mechanism of selenocysteine synthesis on tRNASec in mammals was previously studied by means of HSe- as a Se donor to synthesize selenocysteine. It has been recently established that HSe- in E. coli is activated by ATP to become selenophosphate (SeP). In this study, we provide evidence that [75Se]selenocysteine is produced by bovine selenocysteine synthase from Ser-tRNASec and [75Se]Sep, synthesized from elemental 75Se and Tris(trimethylsilyl)phosphite. We also studied the stability of SeP by NMR measurement. SeP was stable during storage under nitrogen at -80 degrees C for 3 months in 0.2 M Hepes buffer at pH 6.8. However, SeP decomposed at 0 degree C in air (half life 32 hrs) or at 22 degrees C under nitrogen (half life 30 hrs) at pH 6.8. The half lives of SeP at -19 degrees C in air and at 0 degree C under nitrogen at pH 6.8 were 740 and 840 hrs, respectively. At pH 4 under nitrogen at 22 degrees C, the half life was 240 hrs. The half life was only 9.2 hrs at pH 9 under nitrogen at 0 degree C. Thus, SeP was proved to be stable at low temperature, under acidic and anaerobic conditions, but labile under neutral and alkaline conditions. The LD50 of SeP administered i.p. to mice was 37.5 mg/kg body weight.
Se is an essential trace element and is found as a selenocysteine in the active site of Se-enzymes, such as glutathione peroxidase. tRNASec is first aminoacylated with serine by Ser RS and further is converted to selenocysteyl-tRNA by selenocysteine synthase. Mammalian selenocysteine tRNA has dual identities with Ser RS and selenocysteine synthase. Key identity elements for selenocysteine synthase are the long 9 bp AA- and long 6 bp D-stems. Major serine tRNA was converted to a mutant with a 9 bp AA-stem and 6 bp D-stem, instead of a 7 bp AA-stem and 3 bp D-stem. This mutant was active for selenylation as well as serylation. The relative kinetic parameter (Vmax/Km) of the mutant was 0.052 of the value (1.00) of wild-type Sec tRNA. This low value suggests that there is an unknown fine base specific for selenocysteine synthase. For serylation, mutant having 12 bp and wild type tRNASec having 13 bp of the total length of AA- + T-stems were active but the mutants having 11 or 14 bp were inactive. This shows that SerRS measures the distance between the discrimination base and long extra arm for recognition of tRNASer.
We present a new selenium (Se) map of Japan in more detail than in our previous report (J. Health Sci., 42, 360-366). It contains 150 measured points, especially in the northeast of Japan. In the new sampling points, we did not find high Se levels and most of the measurements were generally at values below 1 mg/ kg. In the Se map of Japan, there are two particularly high measurements, at Mt. Zao (10 mg/kg) and Tateyama Murodo Jigokudani (148 mg/kg). These two areas are the origin of a minor branch of the Abukuma River and the Jouganji River, respectively. We also analyzed the Se levels near two major copper mines, Asio and Besshi. We detected high Se content in rocks at both copper mines and an average Se level in soil from the area near the Asio copper mine. The Se level in soil at Besshi was slightly higher than that at the area near the Asio copper mine. We also measured the Se levels of some pyrites in Japan. The Se levels of most pyrites were some ten mg/kg higher than the general Se level in soil. This study provides information about the fundamental background values of Se levels in soil and rocks to correlate with Se pollution if it occurs in the future.
We measured the amount of Se in bovine liver tRNA. tRNA was chromatographed on a BD-cellulose column and Se-rich tRNA was eluted from the column in front of a main tRNA peak. There was 0.3 mmol Se/mol of tRNA and this level is about one tenth that of Escherichia coli tRNA. This suggests the presence of an enzyme that modifies tRNA with Se in bovine liver. We isolated the activity of this enzyme (selenouridine synthase) by chromatography of bovine liver extracts on a DEAE-cellulose column. ATP and selenophosphate synthetase, as well as selenouridine synthase and tRNA, were necessary for the reaction. 75Se was used to label the reaction products, which were analyzed by TLC after digestion with ribonuclease T2. The position of the 75Se-nucleotide on a TLC plate was identical to that of the Se-nucleotide, 5-methylaminomethyl-2-seleno-Up, prepared from 75Se-tRNA in E. coli.
Background and objectives It has been postulated that air-borne fine water particles (or mist) can induce asthma attacks in asthmatic children. To date, no attempt has been made to quantify the density of air-borne fine water particles with the aim of relating particle density to the etiology of asthma among children. The aim of this study was to investigate the relation of asthma attack frequency and the particle density evaluated in terms of light transmittance. Methods The density of fine water particles was quantified by measuring reductions in light transmittance at 250, 365 and 580 nm at an outdoor location when the surroundings were in darkness. The measurements were made at distances varying from 1 to 3 m from the light sources and performed every morning and evening for 1 year. Each day was separated into two half-day units [i.e., morning (from midnight to noon) and afternoon (from noon to midnight)]. The number of asthma attacks among 121 enrolled asthmatic children was counted for each unit. A possible correlation between the transmittance reduction and frequency of asthma attacks was assessed. Results A significant difference was observed in the extent of reduction in light transmittance at 365 nm between the units with asthma attacks and those without attacks. Furthermore, the reduction in the transmittance was more evident when more asthma attacks were recorded among the patients. No difference was detected in the reduction in light transmittance at 250 or 580 nm. Conclusions These results support the hypothesis that airborne fine water particles are among the etiological factors that induce asthma attacks in asthmatic children.
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