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Biomedical subjects

S Hisamatsu

Publications and source records attributed to S Hisamatsu.

At least 19 recordsLinked to original sources

Characterization of several amino acid transports and glutamine metabolism in MOLT4 human T4 leukemia cells.

The transport system responsible for glutamine, alanine and glutamate in MOLT4 human T4 leukemia cell line were characterized. Kinetic studies of sodium-dependent glutamine and alanine transport exhibited a single saturable high-affinity carrier with a Michaelis constant of 152 +/- 26 microm and 203 +/- 36 microm and a maximal transport velocity of 960 +/- 165 and 1096 +/- 208 nmol/10(9)cells/min, respectively. Glutamate uptake was less than one-tenth of glutamine and alanine, and linearly increased with glutamate concentration which was mediated by diffusion. 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid (SITS), known as anion channel blockers, inhibited the sodium-dependent glutamine and alanine transport by 40% at 10 microm. Cellular contents of these amino acids in MOLT4 cells revealed glutamate to be the highest among them despite low glutamate influx. A glutamine metabolism study using whole cells indicated this high conversion rate from glutamine to glutamate, but no conversion to another amino acid. Based on these results, the high glutamate concentration in MOLT4 was speculated to be synthesized from transported glutamine by active glutaminase.

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfo↗

222Rn concentrations in greenhouses in Aomori Prefecture, Japan.

Greenhouses are possible places with high 222Rn concentrations, since soil, the source of 222Rn, is directly exposed inside them. To examine this point, 222Rn concentrations in 28 greenhouses at five locations in Aomori Prefecture were measured for approximately 1 year with passive Rn detectors. For 1 week, measurements of 222Rn concentration and working level were also carried out with active detectors to get equilibrium factors and the ratio of 222Rn concentration in working time to non-working time in selected greenhouses. The geometric mean of annual 222Rn concentrations in greenhouses was 13 Bq m-3, and the same as that in dwellings and lower than that in indoor workplaces in the prefecture. However, variation of the 222Rn concentrations was far larger than in other environments, and ranged from the lowest level in outdoor workplaces to the highest level in indoor workplaces. Significant seasonal variation was also observed in 222Rn concentrations. The mean effective dose from 222Rn and its progenies was estimated to be 0.047 mSv year-1 for a farmer working in a greenhouse.

Air Pollutants, Radioactive↗

Radon concentration in indoor occupational environments in Aomori Prefecture, Japan.

The (222)Rn concentrations in indoor workplaces were measured in Aomori Prefecture, Japan, and the results are reported here. This survey was part of a program to measure background natural radiation dose rate in the prefecture where the first Japanese nuclear fuel cycling facilities are now under construction. The survey of the (222)Rn concentrations in indoor workplaces was carried out at 107 locations from 1996 to 1998. The (222)Rn concentrations were measured for approximately one year at each site with passive Rn detectors, which used a polycarbonate film for counting alpha-rays and could separate concentrations of (222)Rn from (220)Rn. Weeklong measurements of (222)Rn concentration and working level were carried out with active detectors to get the ratio of (222)Rn concentration during working hours to non-working hours as well as equilibrium factors in selected locations. Diurnal variation of (222)Rn concentration depended on building structure, air-conditioning, time of day and day of the week (week days or weekend). The (222)Rn concentration during working hours was generally lower than that in non-working hours. Although the annual average (222)Rn concentration in indoor occupational environments was higher than that in dwellings, radiation dose for Aomori Prefecture residents from Rn in the former was 14% of the total indoor dose by Rn because of the lower concentration in working hours and lower occupancy factor.

Air Pollution, Indoor↗

Rice uptake and distributions of radioactive 137Cs, stable 133Cs and K from soil.

The distributions of radionuclides in plant components as to radionuclide transfer to animals are important for understanding the dynamics of radionuclides in an agricultural field. Most of the non-edible parts in these components are returned to the soil as organic fertilizer where they may again be utilized in the soil-plant pathway and/or are mixed with feed for livestock. Rice plants were grown in an experimental field and separated at harvest into different components, including polished rice, rice bran, hull, leaves, stem and root, and then the distributions of radioactive 137Cs, stable 133Cs and K in these components were determined. The distribution of 137Cs in polished rice and rice bran was similar to that of 133Cs, while that of K was different. The concentration ratios of 133Cs/K in leaf blade positions increased with aging, which means that the translocation rate of 133Cs in rice plants was slower than that of K. At harvest the distribution of dry weight in polished rice to entire rice plants was 34%, and the distributions of 133Cs in the polished rice and the non-edible parts were 7 and 93%, respectively, whereas those of K in the polished rice and the non-edible parts were 2 and 98%, respectively. Findings suggest that the transfer and distribution of 133Cs, not of K, provide better information on the long-term fate of 137Cs in an agricultural environment.

Agriculture↗

Free water 3H concentration in diet samples collected from 1969 to 1988 in Akita, Japan.

Fallout 3H concentrations in diet samples collected from 1969 to 1988 in Akita Prefecture are reported. Since the data for samples from the 1960's and 1970's are important to understand the long-term movement of 3H in the environment, we searched for old diet samples. Recently, diet samples collected in Akita Prefecture during 1969-1982 were found and subjected to 3H analysis. The samples were originally gathered for nutrition survey programs and had been stored in a refrigerated warehouse at -20 degrees C until we located them. The free water 3H concentrations in the samples were measured and are reported here. The present results are combined with already published data; together they show 3H concentrations exponentially decrease from 1969 to 1988 with an apparent half time of 5.9 y. Levels of diet free water 3H concentrations are also similar to measured and estimated precipitation 3H concentrations in Akita City.

Diet Surveys↗

Organically-bound 3H concentration in rice around atomic energy facilities.

Rice samples collected around atomic energy facilities in the Tokai area were analyzed for organically-bound 3H (OBT). Measurement results were compared with those for samples from control areas in Japan and the People's Republic of China. OBT concentrations of rice in the Tokai area were found to be 4 times higher than the control area as maximum. Although this relatively high 3H concentration shows the effect of 3H release from the facilities into the environment, the concentration level is comparable with those in the rice sample from a northern interior region of China. The OBT concentration of rice sample from the general environment in interior China was 4.5 times higher than the control area in Japan. The radiation dose from the increasing 3H level is negligible in comparison with that from natural radiation.

Air Pollutants, Radioactive↗

Tritium concentrations in pine needle, litter and soil samples.

Samples of pine needle, litter and soil samples collected in/around Akita City and Rokkasho Village in 1989 were analyzed for both free water 3H (FWT) and organically-bound 3H (OBT). The FWT concentrations decrease in the order, litter or soil > pine needle. FWT concentrations in soil depend on the moisture content, and tend to increase with decreasing soil moisture content. This relationship is consistent with the observation that FWT in the soil increases with oxidation of atmospheric tritiated hydrogen gas (HT) and decreases with rainwater dilution. The OBT concentrations increase in the order pine needle < litter < soil at most of the sampling locations. This suggests that historically high soil 3H concentrations may be reflected as high OBT concentrations in soils of the present.

Environmental Monitoring↗

In vitro activity of the hairpin ribozyme derived from the negative strand of arabis mosaic virus satellite RNA.

The negative strand of the satellite RNA of tobacco ringspot virus [(-)sTRSV] is a self-cleaving RNA, of which self-cleaving domain is called the hairpin ribozyme. The negative strand of the satellite RNA of arabis mosaic virus [(-)sArMV] has been suggested to have a hairpin ribozyme-like secondary structure, and we have previously shown that this hairpin domain of (-)sArMV has ribozyme activity. Here we report characterization of the cleavage reaction of the (-)sArMV hairpin ribozyme. Mutagenesis analyses in a trans-acting system revealed, surprisingly, that the wild-type ribozyme was less active than almost all the other mutant ribozymes tested. In a cis-acting system (self-cleaving reaction), however, the reaction of the RNA containing the wild-type sequence proceeds highly efficiently. This result suggests that the inefficient cleavage of the wild-type substrate in trans-acting system may be due to low efficiency at the substrate-binding step but not at the chemical cleavage step in the reaction. We also constructed a chimeric ribozyme between the catalytic hairpin domain from (-)sArMV and the substrate-binding site from (-)sTRSV. This chimeric ribozyme had the highest activity among the trans-acting hairpin ribozymes tested.

Base Sequence↗

Tritium concentrations of blood samples collected throughout Japan.

Tritium concentrations were measured for blood samples collected from 20 cities throughout Japan during 1989-1990. The mean 3H concentration was found to be 1.4 +/- 0.4 Bq L-1 and 1.0 +/- 0.4 Bq L-1 (combustion water) for free water 3H and organically-bound 3H, respectively, excluding the abnormally high data of one city. The organically-bound 3H contents clearly depended on the latitudes of sampling locations, although the free water 3H concentrations showed no correlation with the latitudes. Organically-bound 3H is considered to be more suitable than free water 3H as an indicator of long time 3H exposure to human.

Environmental Exposure↗

Hairpin ribozyme-mediated cleavage of the full-length beta-glucuronidase (GUS) mRNA.

We designed three hairpin ribozymes to cleave Escherichia coli beta-glucuronidase (GUS) mRNA and tested those activities in vitro. One of the ribozymes was designed to form 9 base pairs in total with the target GUS mRNA, and the other two ribozymes had longer substrate binding sites. All ribozymes cleaved the model substrate (100 bases long) at the predicted target site. Two ribozymes containing longer substrate binding sites cleaved the substrate much more efficiently than the other ribozyme containing shorter substrate binding site. Also, the ribozymes with long substrate binding sites had high activity against the full-length GUS mRNA (1.9 kilobases) and maintained the activity even at a low temperature, 26 degrees C, a general growth condition of plant cells. Effects of the substrate binding site length of the ribozyme on cleavage activity are discussed.

Base Composition↗

Cis and trans reactions of hairpin ribozymes derived from the negative strand of arabis mosaic virus satellite RNA.

We have previously shown that the hairpin ribozyme-like structure of the negative strand of the satellite RNA of arabis mosaic virus [(- )sArMV] has indeed ribozyme activity. However, some mutagenesis analyses revealed surprisingly that the wild type ribozyme was less active than almost all the other mutant ribozymes tested. These results were derived from a trans-acting system. Here we tested this ribozyme activity in a cis-acting system. We show that the (-)s ArMV hairpin ribozyme has different target-site specificities between cis and trans cleavages. The wild type ribozyme has the highest self-cleaving activity among the ribozyme variants tested.

Base Sequence↗

Acquirement of hairpin ribozyme activity by the long substrate-binding site.

Three hairpin ribozymes were designed to cleave Escherichia coli beta-glucuronidase (GUS) mRNA at a target site. Two of the designed ribozymes (HG10L and HG10L2) had long substrate binding sites and the other (HG10) had short substrate binding site. All three ribozymes cleaved the substrate, and HG10L and HG10L2 cleaved it efficiently. However, HG10 had very low activity. Effect of length of substrate binding site of the ribozyme will be discussed.

Base Sequence↗