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The relationship between donor cell cycle and the developmental ability of somatic cell nuclear transfer (SCNT) embryos has not fully been elucidated. Donor cells that are usually prepared by serum starvation or confluent-cell culture for SCNT represent a heterogeneous population that includes mainly G0 phase cells, other cells in different phases of the cell cycle and apoptotic cells. In this study, we compared the developmental ability of porcine SCNT embryos reconstructed from G0 phase cells (G0-SCNT embryos) and strictly synchronized-G1 phase cells (G1-SCNT embryos), and examined the developmental rates and timing of first DNA synthesis. The G0 phase cells were synchronized by confluent culture, and the G1 phase cells were prepared from actively dividing M phase cells. The G1-SCNT embryos showed a significantly higher (P<0.05) developmental rate to the blastocyst stage per cleaved embryo (59%) than the G0-SCNT embryos (43%). Moreover, initiation of first DNA synthesis and cleavage occurred significantly earlier in the G1-SCNT embryos than in the G0-SCNT embryos. Delay of initiation of first DNA synthesis in the SCNT embryos by aphidicolin resulted in decreased developmental rates to the blastocyst stage without any effect on cleavage rates. Our data demonstrates that synchronized-G1 phase cells can be used as donor cells for SCNT embryos and that earlier initiation of first DNA synthesis may be important for subsequent development of SCNT embryos. The SCNT system using G1-synchronized cells, in terms of their highly uniform and viable cell states, can be useful for studying the reprogramming processes and embryonic development of SCNT embryos.  相似文献   
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Abstract

Emission rates of dimethyl sulficle (DMS), carbonyl sulfide (COS), and carbon disulfide (CS2) to the atmosphere from paddy fields at Ryugasaki, Ibaraki Prefecture, Japan were measured by using the closed chamber method. DMS was emitted mainly through the rice plant and its emission rate was much higher than those of COS and CS2. During a cropping period COS was slightly absorbed by the rice plant. Significant diurnal and seasonal variations of DMS iuxes were observed. The highest DMS iux was observed immediately after the heading day of rice plant. The annual DMS emission rate was in the order of mineral plot > straw plot > no-N plot, ranging from 4.5 to 6.9 mg S m-2 yr-1. The annual COS emission rate was in the order of straw plot > no-N plot > mineral plot, ranging from -0.2 to 1.8 mg S m-2 yr-1. The annual CS2 emission rate was slightly higher in the straw plot, ranging from 0.9 to 2.0 mg S m-2 yr-1.  相似文献   
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The meat quality of farmed yellowtail Seriola quinqueradiata fed on extruded pellets (EP) containing 0.5% (v/v) red pepper (experimental group) was compared with yellowtail of the same age fed on EP (control group). In 1-year-old yellowtail, the crude lipid content of the dorsal muscle of the experimental group tended to be lower than that of the control group. In contrast, there was no difference in the lipid content of the dorsal muscle between the control group and the experimental group in 2-year-old yellowtail. The muscle texture of the experimental group was significantly firmer than that of the control group, with the effect of red pepper unrelated to fish age and lipid content. Color change of red muscle of the experimental group was significantly lower than that of the control group, and the content of thiobarbituric acid-reactive substances in the red muscle was significantly lower in the experimental group than in the control group. These results are the first to demonstrate that the inclusion of red pepper in the diet is able to reduce the loss of muscle texture firmness and to slow down color change in red muscle of yellowtail.  相似文献   
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Lancefield group C Streptococcus dysgalactiae (GCSD) causes severe necrotic lesions in the caudal peduncle in the genus Seriola farmed in Japan. To develop a sero‐diagnostic method for GCSD infection in farmed fish, we attempted to identify a surface immunogenic protein that induces an antibody after infection with GCSD by immunoblot analysis using sera collected from infected fish. A protein obtained from sodium dodecyl sulfate (SDS) extracts of GCSD was identified as S. dysgalactiae surface immunogenic protein (Sd‐Sip). Sd‐Sip exhibited more than 94% homology with a surface antigen or a hypothetical protein from S. dysgalactiae mammalian isolates at the nucleotide sequence level. Expression of the recombinant Sd‐Sip (rSd‐Sip) was confirmed by immunoblot analysis, that is, its reactivity to GCSD‐infected sera. Antibody detection ELISA using rSd‐Sip and their usefulness for diagnosis of GCSD infection were examined. GCSD‐infected sera collected from farmed amberjack, Seriola dumerili (Risso), showed strong reaction with immobilized rSd‐Sip. Meanwhile, sera immunized by other pathogenic bacteria of fish were showed ELISA values similar to those of non‐infected sera. These results of this study suggest that the antibody detection ELISA using rSd‐Sip is an effective diagnostic method for GCSD infection in fish.  相似文献   
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An automatic counting method was developed for fish blood cells using a fluorescent dye, 3, 3-dihexyloxacarbocyanine (DiOC (6)(3)), that selectively stain lipid bilayers in living cells. In the present study, the DiOC(6)(3) method was applied to quail (Cotumix cotumix japonica) blood cells. After quail blood cells were stained with DiOC(6)(3), absolute counts and relative proportions of erythrocytes, granulocytes, monocytes, and lymphocytes plus thrombocytes in whole blood were obtained by means of flow cytometry (FC). The number of each cell types by the FC was in good agreement with those counted microscopically. This method will offer new possibilities for routine blood cell counting for avian medicine.  相似文献   
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There have been diverse reports on the effects of diazepam on cardiac contractility. The purpose of this study was to examine whether diazepam modifies the inotropic response elicited by histamine on an isolated guinea-pig papillary muscle. The responses of electrically driven papillary muscle to histamine and cyclic AMP-related inotropic agents were recorded in the absence and in the presence of diazepam. Histamine and forskolin, which directly stimulate adenylate cyclase, significantly increased the contractile force in the papillary muscle in a concentration-dependent manner. A histaminergic H2-receptor antagonist, cimetidine, but not a H1-receptor antagonist, diphenhydramine, at 10 μ M produced a rightward shift in the concentration-response curve for histamine. Diazepam (10 μ M ) shifted the concentration-response curve for histamine and forskolin to the left by 1.8 and 1.6 times, respectively. Neither a central type (fulmazenil) nor a peripheral type (PK11195) of benzodiazepine receptor antagonist modified the effect of diazepam on the histaminergic-evoked contraction. Phosphodiesterase blockade by 3-isobutyl-1-methylxanthine shifted the concentration-dependent curve for histamine to the left. A combination of 3-isobutyl-1-methylxanthine also produced a leftward shift of the curve. However, there was no significant difference between the 3-isobutyl-1-methylxanthine only group and the combination group. These results indicate that diazepam potentiates the positive inotropic effect produced by histamine, probably mediated via an increase in cyclic AMP levels induced by histamine.  相似文献   
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