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1.
This study was carried out to investigate the effects of minimum essential medium (MEM) vitamins during in vitro maturation (IVM)/in vitro culture (IVC) of porcine nuclear transfer (NT) embryos on subsequent developmental capacity in vitro. Porcine cumulus-oocyte complexes (COCs) were divided into five groups, matured for 44 h in maturation medium with various concentrations of MEM vitamins (0, 0.05, 0.1, 0.2 and 0.4%), and observed for maturation rate. Also, COCs were matured in NUSU-23 media without MEM vitamins for 44 h and cultured in PZM-3 media with various concentrations of MEM vitamins (0, 0.05, 0.4 and 1.0%) for 6 days following nuclear transfer. Factorial (IVM/IVC) experiments were also performed in NCSU-23 medium with or without 0.05% MEM vitamins and PZM-3 medium with or without 0.4% MEM vitamins. They were then tested by examining in vitro development of the porcine reconstructed embryos. The maturation rates of the COCs treated with the MEM vitamins did not differ significantly among the MEM vitamin-treated groups. Addition of vitamins to culture medium did not affect development of porcine reconstructed embryos in vitro. However, addition of low concentrations of MEM vitamins only to maturation medium increased (P<0.05) the proportion of NT embryos developing into blastocysts compared with the control group. Addition of MEM vitamins to IVC medium did not enhance the developmental rate compared with the control group. Thus, addition of MEM vitamins to IVM medium could improve subsequent blastocyst development of porcine NT embryos.  相似文献   
2.
The pharmacophore of the neonicotinoid insecticide imidacloprid, nitroiminoimidazolidine, was modified to heterocycles such as thiazolidine, pyrrolidine, dihydroimidazole, dihydrothiazole, and pyridone conjugated to nitroimine (=NNO2) or nitromethylene (=CHNO2). Their 6-chloro-3-pyridylmethyl or 5-chloro-3-thiazolylmethyl derivatives were examined for insecticidal activity against the American cockroach by injection and for neuroblocking activity using the cockroach ganglion. Most of the compounds having the neonicotinoidal pharmacophore exhibited insecticidal activity at the nanomolar level, which was enhanced in the presence of synergists, and high neuroblocking activity at the micromolar level. Quantitative analysis for the compounds showed that the neuroblocking potency is proportional both to the Mulliken charge on the nitro oxygen atom and to the partition coefficient log P value. The equation for the insecticidal versus neuroblocking potencies indicated that both potencies are related proportionally with each other when the other factors are the same.  相似文献   
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We introduced a fusion gene of human albumin and enhanced green fluorescent protein (EGFP) into porcine oocytes using the sperm vector method, and produced a piglet that showed clear expression of GFP in the hooves and skin. PCR and Southern blotting analysis of genomic DNA extracted from the piglet's tissues, including the liver, showed that the tissues carried the transgene. RT-PCR analysis demonstrated that both the human albumin and EGFP genes were expressed in the tissues. The fact that human albumin gene was integrated and expressed in the liver of the transgenic pig opened a way for us to achieve our goal, which was the use of transgenic pigs for the bioartificial liver support system.  相似文献   
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ABSTRACT:   Restriction fragment length polymorphism analysis was performed on polymerase chain reaction-amplified DNA fragments containing the D-loop , ND2 , and CO I genes of fiddler crab Uca arcuata mitochondrial DNA. In total, 316 individuals from six populations in Japan and two populations in Taiwan were analyzed using five restriction endonucleases ( Afa I, Bcn I, Cfr 13I, Hae III and Hin fI), yielding 85 haplotypes. Samples were taken from Nakagusuku Bay, Okinawajima Island, which is the only known distribution of U. arcuata in the Ryukyu Archipelago. The Okinawajima Island population is isolated geographically from others and showed a marked low genetic variability ( h  = 0.2539, π  = 0.0005) and significant differentiation from other population samples in haplotype composition. We suggest that a substantial decrease in the genetic variability of the Okinawajima Island population was caused by genetic drift under the conditions of small population size and low gene flow from other populations. It is important to conserve the intertidal zone in Nakagusuku Bay for the maintenance of this endangered population.  相似文献   
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In the production of cattle nuclear transfer embryos, the production efficiency is affected by the oocyte developmental competence and successful enucleation rate. This study investigated the effect of treating oocytes with milrinone, a phosphodiesterase inhibitor, on these two characteristics. When cumulus-oocyte complexes (COCs) were cultured for 19 h with 0, 50 or 100 μM of milrinone, the enucleation rate was significantly improved by 100 μM milrinone. However, milrinone treatment during in vitro maturation (IVM) also delayed meiotic progression by at least 2 h, which would affect the examination of enucleation rate and developmental competence of oocytes. Thus, in the second experiment, meiotic resumption was temporarily inhibited with butyrolactone I (BL-I; 100 μM, 18 h) to decrease the delayed maturation caused by milrinone; this enabled a more accurate comparison of the effects of milrinone after oocyte maturation. In nuclear transfer embryo production, oocytes treated with milrinone (100 μM, 20 h) showed a significantly higher rate of enucleation compared with that of control oocytes. This improved enucleation rate was associated with a closer location of the metaphase plate to the first polar body in the treated oocytes compared with that in control oocytes. Furthermore, milrinone improved the frequency of development to the blastocyst stage in the resulting embryos. In conclusion, milrinone supplementation during IVM improved enucleation rates by rendering the metaphase plate in close proximity to the first polar body, and this treatment also improved oocyte developmental competence. These benefits additively improved the yield of cloned embryos that developed to the blastocyst stage.  相似文献   
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Fine- to medium-textured soils are distributed on the Early Pliocene rocks and Pleistocene sandy sediments in the Ka-Etsu plateau. The Jingaoka (JIN) soil was classified into Aluandic Andosols by WRB, and Low-humic Non-allophanic Kuroboku soils by the Fourth Committee for Soil Classification and Nomenclature of the Japanese Society of Pedology, although it was similar morphologically to Yellow-Brown Forest soils. On the other hand, the Tomitsu (TMI) and Yamamuro (YAM) soils were classified into Hyperdystri-Chromic Cambisols by WRB, and Typic Yellow-Brown Forest soils by the Fourth Committee for Soil Classification and Nomenclature based on the morphological, physical and chemical properties. These soils were influenced considerably by the Akahoya tephra, especially the JIN soil. Based on the results of clay-mineralogical analysis and the measurement of the ESR-signal intensity associated with the oxygen vacancies in fine quartz, a large part of parent materials in the soils distributed on the Ka-Etsu plateau was derived from the eolian dust which was brought with the NW winter monsoon from the Precambrian sediments in the northern part of the Asian continent during MIS 2 (24–11 thousand years ago) at the Last Glacial Maximum. The parent materials of the fine-textured JIN soil located on the highest terrace near the rocky coastline were mostly composed of eolian dust, although the influenced of the Akahoya tephra was recognized. The parent materials of the TMI soil were admixed with a small amount of autochthonous materials from the sandy coast located windward. In the YAM soil which was located on the side of a hilly area distant from the coastline, the ESR-signal intensity in fine quartz was considerably low, and the admixture of autochthonous materials was also considered since the texture of this soil was coarser than that of the JIN and TMI soils.  相似文献   
9.
海栖热袍菌极端耐热木聚糖酶B的提纯   总被引:6,自引:0,他引:6  
克隆并在大肠杆菌中表达的海栖热袍菌的 xyn B基因 ,其表达产物木聚糖酶 B的 C 末端带有 6×His标签 ,研究了这种基因重组酶的提纯方法。通过对粗酶提取液的热变性处理 ,Ni NTA亲和柱层析和离子柱层析 ,最终得到了电泳纯的木聚糖酶 B,提纯倍数 4 4.4 ,得率 11。SDS PAGE法测定木聚糖酶 B的相对分子质量为 4 2 ku,与理论推算值 4 2 333u相吻合  相似文献   
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