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1.
Mouse pronuclear stage embryos with intact slit zona pellucida (manipulated) were cultured in vitro until the hatched blastocyst stage in simplex optimized medium with higher K+ concentration (KSOM) prepared with three different water types: tap, deionized reverse osmosis (D‐O) water and Milli‐Q system (M‐Q) water. The culture media were supplemented with or without protein and ethylenediaminetetraacetic acid (EDTA, disodium salt). The rates of hatched blastocysts were significantly affected (p < 0.01) by micromanipulation, protein supplement and water source. The water source has no influence (p > 0.05) on development in EDTA‐supplemented protein‐free culture media, whereas in EDTA‐ and protein‐free culture media, the water quality significantly (p < 0.001) affected the rates of development, with higher rates in media prepared with M‐Q water. The micromanipulated embryos showed higher sensitivity to the water quality (p < 0.01). It worth mentioning that the rates of hatched blastocysts in protein‐free culture media were very low (0–7.5%). Furthermore, the three different water types were analysed by measuring the electrical conductivity, inorganic ions, total organic carbon and endotoxins to evaluate the purity. M‐Q water showed the lowest levels of inorganic ion, total organic carbon and endotoxin concentrations. We concluded that manipulated mouse embryos are good system to evaluate the quality of water used in biological system. 相似文献
2.
E Ogawa A Kawakami T Yagi T Amaya H Fujise R Takahashi 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》1992,54(1):57-62
Oxidative damage to the membrane in canine erythrocytes with inherited high Na, K-ATPase activity (HK cells) was compared with that in normal canine cells (LK cells). When 30 mM beta-acetylphenylhydrazine (APH) was applied to HK and LK cells, lipid peroxidation and hemoglobin denaturation occurred. Lipid peroxidation determined from malondialdehyde (MDA) formation was significantly lower in HK than in LK cells so far as endogenous glutathione (GSH) concentration was maintained at appropriate levels. With the depletion of GSH, MDA formation was accelerated and difference between HK and LK cells was not significant. Denatured hemoglobin bound to the membrane protein was less in HK than in LK cells. During incubation with APH, osmotic fragility increased markedly in LK cells, while HK cells showed very little change. The amounts of total lipid, total and free cholesterol, glycolipid, phospholipid and fatty acids were essentially the same in both cell types. Fatty acid compositions showed very small differences. The membrane of HK cells thus appear to have greater protection against oxidative damage induced by APH, owing to the presence of excess GSH in HK cells. The capability of HK cells to withstand oxidative damage would not be due to differences in membrane lipid compositions. 相似文献
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Kobayashi S Sakatani M Kobayashi S Okuda K Takahashi M 《The Journal of reproduction and development》2007,53(6):1305-1311
Inhibition of specific gene expression using RNA interference (RNAi) is a valuable tool for functional analysis of a target gene. However, there is little information available concerning RNAi for analysis of gene function in relation to the reproductive physiology of follicular cells in ruminants. Thus, the aim of this study was to evaluate the interfering effect of small interference RNA (siRNA) on expression of cyclooxygenase-2 (Cox-2) mRNA and prostagrandin F(2alpha) (PGF(2alpha)) production in bovine cumulus-granulosa (CG) cells. Bovine CG cells were collected from aspirated follicles and cultured. After reaching confluency, two experiments were conducted. In experiment 1, to investigate the effective concentration of siRNA, 0, 100, 250 and 500 pM of Cox-2 siRNA was introduced into the CG cells, respectively. After 24 h, the amount of Cox-2 mRNA expression was measured by RT-PCR and real-time PCR. In experiment 2, to investigate the time required for effective interference of siRNA and Cox-2 activity, 250 pM siRNA was introduced for 0, 3, 6, 12 and 24 h. After culture, the amount of Cox-2 mRNA expression was measured and the culture medium was collected to determine the PGF(2alpha) concentration by enzyme immunoassay. The Cox-2 mRNA expression was not affected by introduction of 100 pM siRNA into CG cells for 24 h, but 250 and 500 pM Cox-2 siRNA significantly reduced the Cox-2 mRNA expression. Moreover, the significant suppressive effect of 250 pM siRNA was observed 6 h after introduction, and the reduction of mRNA expression by RNAi became more obvious over 12 h. On the other hand, the PGF(2alpha) concentration in the culture medium was not significantly different 12 h after siRNA introduction; however, the PGF(2alpha) concentration 24 h after siRNA introduction was significantly decreased compared with the control at the same time point. These results suggest that gene silencing of Cox-2 with siRNA is capable of analyzing the function and expression of specific genes in bovine CG cells. 相似文献
7.
Seiko Yoshikawa Hidehiro Takahashi Yasuko Sasada Hidetoshi Mochizuki 《Soil Science and Plant Nutrition》2013,59(6):898-909
AbstractThe aim of this study was to evaluate the impact of land use on nitrate nitrogen (NO3-N) in shallow groundwater (G-N) and total nitrogen (N) in river water (R-N). The study area consisted of 26 watersheds (1342 km2) covering 72% of Kagawa Prefecture in Japan. We estimated G-N specific concentrations, which showed the magnitude of the upland fields, paddy fields, forests and urban land-use contributions to watershed-mean G-N. G-N specific concentrations were gained as partial regression coefficients using a multiple regression analysis of the watershed-mean G-N concentrations and the land-use ratios in each of the 26 watersheds. The results showed that the G-N specific concentration, which was gained as the partial regression coefficient for the multiple regression analysis, was 15.2 mg L?1, 10.3 mg L?1, 2.3 mg L?1 and 2.5 mg L?1 for the upland fields, paddy fields, forests and urban land-use types, respectively. R-N pollution load runoff to the river mouth was calculated by multiplying R-N specific concentration (previously reported) by river flow at the river mouth. Similarly, G-N pollution load arrival to groundwater was calculated by multiplying G-N specific concentration by the groundwater flow. The R-N pollution load runoff was 19.3 kg ha?1 y?1, 7.7 kg ha?1 y?1, 1.7 kg ha?1 y?1 and 7.6 kg ha?1 y?1, while the G-N pollution load arrival was 7.3 kg ha?1 y?1, 5.0 kg ha?1 y?1, 1.1 kg ha?1 y?1 and 1.2 kg ha?1 y?1, for upland fields, paddy fields, forests and urban areas, respectively. These results showed that the N in river water and groundwater was derived mainly from runoff and leaching from croplands. Therefore, the relationships between watershed-mean non-absorbed, applied nitrogen (NAA-N: nitrogen applied to cropland via fertilizer and manure without being absorbed by crops), R-N concentration and watershed-mean G-N concentration were investigated. A curvilinear correlation was observed between NAA-N and R-N concentrations (r2 = 0.68) except for one small, high-density, urban watershed, and a weak linear correlation was observed between NAA-N and G-N concentrations (r2 = 0.42). 相似文献
8.
Arata Katayama Hong-Ying Hu Mamie Nozawa Sayuri Takahashi Koichi Fujie 《Soil Science and Plant Nutrition》2013,59(6):841-846
Based on the respiratory quinone profile, changes in the structure of microbial communities in the soil samples from Nagoya University Farm were monitored after the treatment with 1% of a mixture of glucose and peptone. Samples of two soils differing in the fertilization history were examined: CF-soil with the application of only chemical fertilizers and FYM-soil with the application of only farmyard manure at a high rate. In the CF-soil, the amount of water-soluble organic carbon (WOC), indicator of the mixture of glucose and peptone, decreased to the original level after 14 d. After 7 d, the soil pH reached the maximum level, then decreased gradually. Changes in the inorganic nitrogen levels in the water extract also reflected the 14-d period of mineralization. The amount of respiratory quinones reached maximum levels after 7 d and gradually decreased, reflecting the changes in the microbial biomass. The quinone composition significantly changed during the 14-d period and returned to a profile similar to the original one after 28 d. Diversity of quinones significantly decreased during the 14-d period due to the predominance of ubiquinone with 9 isoprenoid units. In the FYM-soil, the amount of WOC decreased to the original level after 1 d, and the pH and inorganic nitrogen levels in the water extract reflected the one-day mineralization period, and nitrification started after 3 d. Although the amount of quinones indicated an increase in the microbial biomass for 14 d, the quinone composition did not change. These findings suggested that long-term application of farmyard manure resulted in stable microbial communities in response to the incorporation of organic matter in soil. 相似文献
9.
Kazuo Satoh Tomohiro Tanaka Yuuichi Oguro Reiji Takahashi Tatsuaki Tokuyama 《Soil Science and Plant Nutrition》2013,59(5):777-781
Long-term preservation of ammonia-oxidizing bacteria had been achieved until now by using the silica gel-freezing method. Recently, the freeze-drying method for the preservation of microorganisms has been widely used, and has been applied to the preservation of ammonia-oxidizing bacteria. The 10% skim milk-l% pyruvate or glutamate ratio was optimum as a dispersion-medium, and the preservation temperature of the freeze-dried cells was effective at -80°C. This method of preservation was found to be considerably more effective than the silica gel-freezing method. 相似文献
10.
Daisuke Takahashi 《Ecology of Freshwater Fish》2008,17(1):71-77
Abstract – Age at maturity in males of the freshwater goby Tridentiger brevispinis , a species with exclusive paternal care, was investigated in two populations in Lake Biwa, Japan, that differed markedly in nest site abundance. At Ohmi-maiko, where nest sites were scarce, most males matured at age 3, and mean body size of males guarding eggs in nests was larger than that of males sampled randomly (including both guarding and nonguarding males) in the population. Conversely, at Minamihama, where nest sites were abundant, many males matured at age 1, and there was no difference in body size between guarding males and males collected randomly. The slope of regression lines between body size of the guarding male and the number of eggs in his nest was greater in Ohmi-maiko than in Minamihama. These results suggest that the shortage of nest sites enhances reproductive success in larger males, probably through male–male competition for nest sites and female mate choice for larger males. I conclude that in nest spawners, the availability of nest sites should strongly affect life-history traits of males through sexual selection. 相似文献