Effect of centrifugation treatment before vitrification on the viability of porcine mature oocytes and zygotes produced in vitro     

Somfai T  Kashiwazaki N  Ozawa M  Nakai M  Maedomari N  Noguchi J  Kaneko H  Nagai T  Kikuchi K 《The Journal of reproduction and development》2008,54(3):149-155

The aim of the present study was to investigate the effects of centrifugation pretreatment on the viability and nuclear status of porcine in vitro matured (IVM) oocytes and on the developmental competence of in vitro fertilized (IVF) oocytes (zygotes) after cryopreservation by vitrification (Solid Surface Vitrification; SSV). Mature oocytes having the first polar body after IVM and zygotes having the second polar body at 10 h after IVF were centrifuged at 10,000 x g at 37 C for 20 min and then subjected to SSV. Their viability was evaluated by morphological appearance and fluorescein diacetate staining. The nuclear status of oocytes was evaluated 6 h after vitrification. The developmental ability to the blastocyst stage of vitrified zygotes was evaluated after 6 days of in vitro culture. Although centrifugation did not damage the oocytes directly, it drastically reduced the rate of live oocytes after SSV. The rates of vitrification-induced parthenogenetic activation were similar in both centrifuged and non-centrifuged oocytes (42.4 and 47.4%, respectively). Centrifugation had no significant effects on the viability of pronuclear oocytes. The development of vitrified zygotes to the blastocyst stage was significantly lower than that of the control irrespective of centrifugation pretreatment. There was no difference in the cleavage and blastocyst rates between the control and centrifuged zygotes after vitrification. There was also no difference in the total cell numbers of blastocysts between the control and centrifuged zygotes irrespective of vitrification. These results reveal that, in IVM porcine oocytes, centrifugation pretreatment is highly detrimental to cryotolerance; however, in zygotes, it has only a slight effect on viability and does not alter the developmental competence of surviving zygotes.  相似文献   

微管相关的细胞型朊病毒蛋白NH2端细胞内定位     

Naomi S.Hachiya  宁章勇  赵德明 《中国畜牧兽医》2004,31(7):22

利用细胞型朊病毒蛋白(PrPc)的双荧光标记,发现PrPc NH2端和COOH端片段在N2a细胞和HpL3～4细胞内有明显不同的分布,N2a细胞是鼠成神经瘤细胞,HpL3～4细胞是prnp基因敲除鼠的海马细胞系细胞.值得注意的是,主要定位于细胞内部分的PrPc的NH2端片段,采用微管解聚剂(nocodazole)处理后在细胞内聚集.鼠PrPc的残基片段1～121、1～111和1～91在细胞内呈现适当的分布轮廓,而残基1～52和1～33不能表现出明显的分布区域,这些资料显示微管相关的PrPcNH2端片段的定位至少需要包含91个氨基残基.  相似文献   

The effect of M‐phase stage‐dependent kinase inhibitors on inositol 1,4,5‐trisphosphate receptor 1 (IP3R1) expression and localization in pig oocytes          下载免费PDF全文

Anucha Sathanawongs  Katsuyoshi Fujiwara  Tsubasa Kato  Masahiko Hirose  Maki Kamoshita  Richard J. H. Wojcikiewicz  Jan B. Parys  Junya Ito  Naomi Kashiwazaki 《Animal Science Journal》2015,86(2):138-147

At fertilization, inositol 1,4,5‐trisphosphate receptor type 1 (IP₃R1) has a crucial role in Ca²⁺ release in mammals. Expression levels, localization and phosphorylation of IP₃R1 are important for its function, but it still remains unclear which molecule(s) regulates IP₃R1 behavior in pig oocytes. We examined whether there was a difference in localization of IP₃R1 after in vitro or in vivo maturation of pig oocytes. In mouse oocytes, large clusters of IP₃R1 were formed in the cortex of the oocyte except in a ring‐shaped band of cortex adjacent to the spindle. However, no such clusters of IP₃R1 were observed in pig oocytes and there was no difference in its localization between in vitro and in vivo matured oocytes. We next tried to clarify which factor(s) regulates IP₃R1 localization, phosphorylation and expression using M‐phase stage‐dependent kinase inhibitors. Our results show that treatments with roscovitine (p34^cdc2 kinase inhibitor) or U0126 (mitogen‐activated protein kinase inhibitor) did not affect IP₃R1 expression or localization in pig oocytes, although the latter strongly inhibited phosphorylation. However, treatment with BI‐2536, an inhibitor of polo‐like kinase 1 (Plk1), dramatically decreased the expression level of IP₃R1 in pig oocytes in a dose‐dependent manner. From these results, it is suggested that Plk1 is involved in the regulation of IP₃R1 expression in pig oocytes.  相似文献   

Effects of intravenous administration of tranexamic acid on hematological,hemostatic, and thromboelastographic analytes in healthy adult dogs          下载免费PDF全文

Efrat Kelmer DVM  MS  DACVECC  Gilad Segev DVM  DECVIM  Victoria Papashvilli  Naomi Rahimi‐Levene MD  MHA  Yaron Bruchim DVM  DACVECC  Itamar Aroch DVM  DECVIM  Sigal Klainbart DVM  DACVECC 《Journal of Veterinary Emergency and Critical Care》2015,25(4):495-501

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Bacillus cereus from the environment is genetically related to the highly pathogenic B. cereus in Zambia     

Hirohito OGAWA  Miyuki OHNUMA  David SQUARRE  Aaron Simanyengwe MWEENE  Takayuki EZAKI  Daisuke FUJIKURA  Naomi OHNISHI  Yuka THOMAS  Bernard Mudenda HANG’OMBE  Hideaki HIGASHI 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2015,77(8):993-995

To follow-up anthrax in Zambia since the outbreak in 2011, we have collected samples from the environment and the carcasses of anthrax-suspected animals, and have tried to isolate Bacillus anthracis. In the process of identification of B. anthracis, we collected two isolates, of which colonies were similar to B. anthracis; however, from the results of identification using the molecular-based methods, two isolates were genetically related to the highly pathogenic B. cereus, of which clinical manifestation is severe and fatal (e.g., pneumonia). In this study, we showed the existence of bacteria suspected to be highly pathogenic B. cereus in Zambia, indicating the possibility of an outbreak caused by highly pathogenic B. cereus.  相似文献   

Optimization of cryoprotectant treatment for the vitrification of immature cumulus-enclosed porcine oocytes: comparison of sugars,combinations of permeating cryoprotectants and equilibration regimens     

Tamás SOMFAI  Nguyen Thi MEN  Junko NOGUCHI  Hiroyuki KANEKO  Naomi KASHIWAZAKI  Kazuhiro KIKUCHI 《The Journal of reproduction and development》2015,61(6):571-579

Our aim was to optimize the cryoprotectant treatment for the preservation of immature porcine cumulus-oocyte complexes (COCs) by solid surface vitrification. In each experiment, the vitrification solution consisted of 50 mg/ml polyvinyl pyrrolidone, 0.3 M of the actual sugar and in total 35% (v/v) of the actual permeating cryoprotectant (pCPA) combination. After warming, the COCs were subjected to in vitro maturation, fertilization and embryo culture. In Experiment 1, trehalose and sucrose were equally effective during vitrification and warming in terms of facilitating oocyte survival and subsequent embryo development. In Experiment 2, when equilibration was performed at 38.5 C in a total of 4% (v/v) pCPA for 15 min, the combination of ethylene glycol and propylene glycol (EG + PG = 1:1) was superior to EG and dimethyl sulfoxide (EG + DMSO = 1:1) in terms of oocyte survival after vitrification and the quality of resultant blastocysts. In Experiment 3, equilibration in 4% (v/v) pCPA for 15 min before vitrification was superior to that in 15% (v/v) CPA for 5 min for achievement of high survival rates irrespective of the pCPA combination used. In Experiment 4, when equilibration was performed in 4% EG + PG for 5 min, 15 min or 25 min, there was no difference in oocyte survival and subsequent embryo development after vitrification and warming; however, the developmental competence of cleaved embryos was tendentiously reduced when equilibration was performed for 25 min. In conclusion, trehalose and sucrose were equally effective in facilitating vitrification, and the optimum pCPA treatment was 5–15 min equilibration in 4% (v/v) of EG + PG followed by vitrification in 35% (v/v) EG + PG.  相似文献   

Developmental capacity and implantation potential of the embryos with multinucleated blastomeres     

Akiyoshi EGASHIRA  Nobuhiko YAMAUCHI  Keiko TANAKA  Chihiro MINE  Hitomi OTSUBO  Masao MURAKAMI  Md. Rashedul ISLAM  Misako OHTSUKA  Naomi YOSHIOKA  Takashi KURAMOTO 《The Journal of reproduction and development》2015,61(6):595-600

The presence of multinucleated blastomeres (MNBs) in embryos is associated with poor developmental competence in assisted reproductive technologies. This phenomenon is observed not only in humans but also in other animal species. The purpose of the present study was to investigate the characteristics of embryos with MNBs (MNB embryos) that could be utilized in embryo transfer. The developmental rate of MNB embryos to the blastocyst stage (50.8%) was significantly lower than that of normal embryos (73.3%) (P < 0.05). The clinical pregnancy rates of fresh embryo transfer (ET) using day 2 or day 3 embryos were significantly lower in MNB embryos (5.1%) compared with normal embryos (24.0%) (P < 0.05). In the case of frozen-thawed ET using a single vitrified/warmed blastocyst, however, the clinical pregnancy rate of MNB embryos was close to that of normal embryos (59.1% vs. 52.8%). Thus, the findings of the present study suggest that the frozen-thawed ET of MNB embryos might improve the potential for implantation followed by successful pregnancy.  相似文献   

Possible involvement of phosphatidylinositol 3-kinase in the maintenance of metaphase II attest in porcine oocytes matured in vitro     

Junya ITO  Masayuki SHIMADA  Naomi KASHIWAZAKI 《Animal Science Journal》2010,81(1):42-47

It has been reported that phosphatidylinositol 3-kinase (PI3K)-protein kinase B (PKB) pathway plays a crucial role in the meiotic resumption and progression to the metaphase II (MII) stage of oocytes. However, the role of this pathway in meiotic arrest at the MII stage (cytostatic activity) is not well understood. In this study the effect of a PI3K inhibitor, LY294002, on the MAPK and p34^cdc2 kinase activities of matured porcine oocytes was examined. After maturation culture, both the MAPK and p34^cdc2 kinase activities in the oocytes were gradually decreased in a time-dependent manner. Although 25 µmol/L LY294002 did not affect either the MAPK or p34^cdc2 kinase activities, 50 µmol/L LY294002 suppressed the PKB phosphorylation and slightly decreased MAPK activity, but not the p34^cdc2 kinase activity. Therefore the effect of 10 µmol/L Ca²⁺ ionophore which was reported as inducing a transient decrease of p34^cdc2 kinase but not MAPK activities, was also examined in LY294002-treated oocytes. By additional treatment with LY294002 after Ca²⁺ ionophore, both the MAPK and p34^cdc2 kinase activities were decreased in a time-dependent manner, concomitantly with improvement of pronuclear formation. Therefore, we concluded that PI3K is involved in the maintenance of MAPK activity in matured porcine oocytes.  相似文献   

The use of an angularis oris axial pattern flap in a dog after resection of a multilobular osteochondroma of the hard palate     

Dicks N  Boston S 《The Canadian veterinary journal. La revue veterinaire canadienne》2010,51(11):1274-1278

An 11-year-old neutered male boxer was presented for treatment of a multilobular osteochondroma of the hard palate. The mass was surgically resected and the hard palate defect was reconstructed using an angularis oris axial pattern buccal mucosal flap. No local recurrence was reported 6 mo after surgery.  相似文献   

Phytotoxicity of GF-120 NF Naturalyte fruit fly bait carrier on sweet cherry (Prunus avium L.) foliage     

DeLury NC  Thistlewood H  Routledge R 《Pest management science》2009,65(1):52-59

BACKGROUND: Six sweet cherry (Prunus avium L.) cultivars were tested with GF-120 with spinosad (0.2 g L(-1) spinosad bait) or without it (blank bait) to understand leaf phytotoxicity observed in the field. RESULTS: Spinosad bait and blank bait did not differ significantly with respect to damage observed. Leaf damage was found almost exclusively at the abaxial (lower) surfaces with the doses (0, 17, 20, 25 or 40%) and cultivars tested. The effects of the blank bait on abaxial surfaces increased from 24 to 168 h, and with dose, in terms of the proportion of droplets (0.00, 0.42, 0.52, 0.75 or 0.94) and area (0.0, 18.7, 23.5, 40.5 or 91.6 mm) burned. In addition, chlorophyll was reduced with increasing dose on abaxial surfaces (SPAD = 44.6, 36.1, 34.1, 31.0, 21.5), but not on adaxial (upper) surfaces (SPAD = 44.6, 44.2, 44.0, 44.8, 44.4). The chlorophyll level in undamaged leaves (adaxial surfaces) differed by cultivar. Cherry leaves were less damaged by a 20% bait application in June (0.26) than in July (0.46) and August (0.50). Incidental insect leaf feeding at bait locations occurred at a low rate and was highest on abaxial bait surfaces. CONCLUSIONS: Applying GF-120 to the adaxial leaf surface, or at doses of 相似文献