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This study was conducted to evaluate the microtubule distribution following control of nuclear remodeling by treatment of bovine somatic cell nuclear transfer (SCNT) embryos with caffeine or roscovitine. Bovine somatic cells were fused to enucleated oocytes treated with either 5 mM caffeine or 150 µM roscovitine to control the type of nuclear remodeling. The proportion of embryos that underwent premature chromosome condensation (PCC) was increased by caffeine treatment but was reduced by roscovitine treatment (p < 0.05). The microtubule organization was examined by immunostaining β- and γ-tubulins at 15 min, 3 h, and 20 h of fusion using laser scanning confocal microscopy. The γ-tubulin foci inherited from the donor centrosome were observed in most of the SCNT embryos at 15 min of fusion (91.3%) and most of them did not disappear until 3 h after fusion, regardless of treatment (82.9-87.2%). A significantly high proportion of embryos showing an abnormal chromosome or microtubule distribution was observed in the roscovitine-treated group (40.0%, p < 0.05) compared to the caffeine-treated group (22.1%). In conclusion, PCC is a favorable condition for the normal organization of microtubules, and inhibition of PCC can cause abnormal mitotic division of bovine SCNT embryos by causing microtubule dysfunction.  相似文献   
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Newcastle disease (ND) caused by virulent Newcastle disease virus (NDV) is a highly contagious viral disease of poultry. Virulent NDVs characteristically have a multibasic amino acid sequence (virulence motif) such as 112RRQKRF117 at the cleavage site of the precusor fusion (F0) protein. The antigenic and immunogenic characteristics of the virulence motif 112RRQKRF117 in the F0 protein of virulent NDVs were investigated. Epitope mapping analysis revealed that a RRQKRF-specific monoclonal antibody 4G2 recognized the KRF section of the motif. A synthetic peptide bearing the RRQKRF motif reacted strongly with sera from virulent NDV (with RRQKRF motif)-infected chickens. These sera also showed reactivity to peptides bearing other virulence motifs (112KRQKRF117, 112RRQRRF117 and 112RRRKRF117) but not an avirulence motif (112GRQGRL117) by ELISA. The synthetic bearing RRQKRF motif reacted with 60% to 91% of sera taken from surviving chickens on ND outbreak farms but not with sera from vaccinated birds, even though most of the sera had antibody to NDV due to vaccination. This indicates that the virulence motif has the potential to differentiate virulent NDV infected birds from vaccinated birds.  相似文献   
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We investigated the effects of hydrogen peroxide on mucous cells and lysozymes in gill tissue of olive flounder. Morphological analysis revealed significant changes in the number of mucous cells after exposure to 300 and 500 mg L ? 1 hydrogen peroxide; however, there was no detectable difference in the 100 mg L ? 1 group. Lysozyme activities declined gradually in the gill mucus, but in contrast activities in serum increased significantly following the completion of treatment. The hepatic c‐ and g‐type lysozyme levels showed significant responses, indicating that the liver is a major lysozyme producer in response to stress condition; however, in the head kidney, there were no significant changes in the g‐type lysozyme level in all the groups, while the only detectable difference with the c‐type gene in the 300 mg L ? 1. While the treatments showed no significant changes compared with the control for either branchial lysozyme, there were significant differences in c‐type lysozyme between the 3rd treatment and 5 day. Taken together, our data indicate that hydrogen peroxide treatment may lead to modulation of the innate immune response of olive flounder; therefore, the application of hydrogen peroxide in a field system will require more attention to dose and timing effects.  相似文献   
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Nitric oxide (NO) produced by endothelial nitric oxide synthase (eNOS) is a fundamental regulator of systemic blood pressure. Ginsenosides from Panax ginseng have been investigated in vitro for the molecular and biochemical mechanisms by which they stimulate NO release in vascular endothelial cells; however, little research has been done to confirm the physiological relevance of these in vitro studies. To address this research gap, the effects of a P. ginseng extract rich in ginsenosides from protopanaxatriol on spontaneously hypertensive rats (SHRs) was examined. Ginseng extract administration stimulated nongenomic Akt-mediated eNOS activation, enhanced NO production, improved vessel wall thickening, and alleviated hypertension in SHRs, confirming the physiological relevance of previous in vitro studies with ginsenosides.  相似文献   
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A pineapple protease, bromelain, was used to improve the dyeing properties of protein fibers such as wool and silk. The optimal condition for the activity of the pineapple protease was about 60 °C at pH 7. The wool and silk were treated with the protease extracted from a pineapple and the K/S values of the dyed wool and silk were measured using a spectrophotometer in order to compare the dye uptake. The protease treatment enhanced the dyeing properties of protein fibers without severe changes in mechanical properties. The surface appearances of protease-treated fibers were observed by microscopy.  相似文献   
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Background: Critically ill horses are susceptible to thrombotic disease, which might be related to increased platelet reactivity and activation. Objectives: To compare the effect of oral clopidogrel and aspirin (ASA) on equine platelet function. Animals: Six healthy adult horses. Methods: Horses received clopidogrel (2 mg/kg PO q24h) or ASA (5 mg/kg PO q24h) for 5 days in a prospective randomized cross‐over design. Platelet aggregation responses to adenosine diphosphate (ADP) and collagen via optical aggregometry, and platelet secretion of serotonin (5HT) and production of thromboxane B2 (TXB2) by ELISA were evaluated. In horses receiving clopidogrel, high‐performance liquid chromatography analysis for clopidogrel and its carboxylic‐acid metabolite SR 26334 was performed. Results: SR 26334 was identified in all clopidogrel‐treated horses, although the parent compound was not detected. Clopidogrel resulted in decreases in ADP‐induced platelet aggregation persisting for 120 hours after the final dose. ADP‐induced platelet aggregation decreased from a baseline of 70.2 ± 14.7% to a minimum of 15.9 ± 7.7% 24 hours after the final dose (P < .001). Collagen‐induced aggregation decreased from a baseline of 93 ± 9.5% to a minimum of 70.8 ± 16.9% 48 hours after the final dose (P < .001). ASA did not decrease platelet aggregation with either agonist. ASA decreased serum TXB2 from a baseline value of 1310 ± 1045 to 128 ± 64 pg/mL within 24 hours (P < .01). Conclusions and Clinical Importance: Clopidogrel effectively decreases ADP‐induced platelet aggregation in horses, and could have therapeutic applications for equine diseases associated with platelet activation.  相似文献   
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