Bovine OPU‐Derived Oocytes can be Matured In Vitro for 16–28 h with Similar Developmental Capacity     

JS Merton  APW de Roos  EPC Koenen  BAJ Roelen  PLAM Vos  E Mullaart  HM Knijn 《Reproduction in domestic animals》2012,47(6):1037-1042

The aim of this study was to determine the optimal maturation culture period of ovum pick up (OPU)‐derived cumulus oocytes complexes (COCs) in relation to their developmental capacity. Embryo production, embryo cryotolerance, post‐transfer embryonic survival and calf characteristics such as gestation length, birthweight and sex ratio were investigated. This retrospective study covers the analyses of ovum pick up –in vitro production and calving results from a commercial programme that took place between March 1994 and September 2004. Donors were both heifers (of which approximately 90% pregnant) and cows (of which approximately 10% pregnant). Embryo production analyses were based on 7800 OPU sessions conducted from January 1995 until January 1999. Analyses of calving rate were based on 13 468 embryo transfers performed during January 1995 until May 2002. Analyses on calf characteristics were based on 2162 calves born between March 1994 and September 2004. The in vitro maturation culture period ranged from 16 to 28 h. The mean production rate of transferable embryos was 16.5% (1.2 embryos per OPU session). Length of maturation culture period did not affect the production of transferable embryos. Mean calving rate was 40.9% and 38.7% for fresh and frozen/thawed embryos, respectively. Calving rate was not affected by the maturation culture period. Mean birthweight, gestation length and proportion of male calves were 46 kg, 281.9 days and 52.8%, respectively. Maturation culture period did not affect these variables. In conclusion, this study shows that the in vitro maturation culture period within the range of 16–28 h does not affect in vitro embryo production, embryo cryotolerance, post‐transfer embryonic survival and calf characteristics, suggesting that all COC batches collected by OPU on the same day, can be fertilized in one IVF session without a significant loss in the production from oocyte to calf.  相似文献   

Neurogenic and cardiomyogenic differentiation of mesenchymal stem cells isolated from minipig bone marrow     

Kumar BM  Maeng GH  Lee YM  Kim TH  Lee JH  Jeon BG  Ock SA  Yoo JG  Rho GJ 《Research in veterinary science》2012,93(2):749-757

The present study investigated the potential of minipig bone marrow-mesenchymal stem cells (BM-MSCs) to differentiate in vitro into neuron- and cardiomyocyte-like cells. Isolated BM-MSCs exhibited a fibroblast-like morphology, expressed CD29, CD44 and CD90, and differentiated into osteocytes, adipocytes and chondrocytes. Upon induction in two different neuronal specific media, most of BM-MSCs acquired the distinctive morphological features and positively stained for nestin, neurofilament-M (NF-M), neuronal nuclei (NeuN), β-tubulin, galactocerebroside (Gal-C) and glial fibrillary acidic protein (GFAP). Expression of nestin, GFAP and NF-M was further demonstrated by RT-PCR and RT-qPCR. Following cardiomyogenic induction, MSCs exhibited a stick-like morphology with extended cytoplasmic processes, and formed cluster-like structures. The expression of cardiac specific markers α-smooth muscle actin, cardiac troponin T, desmin and α-cardiac actin was positive for immunofluorescence staining, and further confirmed by RT-PCR and RT-qPCR. In conclusion, our results showed the in vitro differentiation ability of porcine BM-MSCs into neuron-like and cardiomyocyte-like cells.  相似文献   

Effects of Developmental Stage, Embryonic Interferon-τ Secretion and Recipient Synchrony on Pregnancy Rate after Transfer of in vitro Produced Bovine Blastocysts     

HM Kubisch  S Sirisathien  P Bosch  HJ Hernandez-Fonseca  G Clements  JR Liukkonen  BG Brackett 《Reproduction in domestic animals》2004,39(2):120-124

Three separate trials of bovine embryo transfers were performed consisting of 32, 41 and 33 transfers, respectively, to examine the effects of (a) the developmental stage of in vitro‐derived blastocysts, (b) the amount of interferon‐τ (IFN‐τ) they secreted during culture and (c) the cyclic stage of the recipient at the time of transfer on the probability of establishment of pregnancy. One blastocyst was transferred into the ipsilateral uterine horn to the CL. At the time of transfer, blastocysts were classified into one of three developmental stages (early blastocyst, blastocyst and expanded blastocyst) and the cyclic stage of each cow was assessed (?12 h, on time, +12 h, +24 h, >24 h). Prior to the second and third trials, blastocysts were individually cultured for 24 h in 50 μl medium droplets and the IFN‐τ concentration in the droplet was determined. Logistic regression analyses revealed that expanded blastocysts had a significantly higher likelihood of establishing pregnancy (p = 0.009), and that there was a significant interaction with the cyclic stage of the recipient in this group with lower rates of pregnancy resulting from decreasing synchrony with the recipient (p = 0.033). IFN‐τ secretion during culture was significantly higher in expanded blastocysts than in the other two groups (p < 0.05). A significant effect of the pre‐transfer level of IFN‐τ secretion was found only in the ‘Blastocyst’ group where transfer of embryos with lower IFN‐τ production prior to transfer resulted in higher pregnancy rates (p = 0.047). These results demonstrate that IFN‐τ secretion may be a useful tool to predict pregnancy outcome, but only within certain developmental stages.  相似文献   

Impact of Growth Factors on In Vitro Development of Caprine Oocytes at Pre-antral Stage     

HM Zhou  Y Zhang 《Reproduction in domestic animals》2005,40(2):161-165

The objective of this study was to examine the effect of various growth factors such as the epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I) either individually or in association, in the presence of follicle-stimulating hormone (FSH) on the in vitro growth and viability of caprine oocytes at pre-antral stage. Pre-antral follicles were disassociated enzymatically and mechanically from pre-pubertal caprine ovaries after the animals were anaesthetically ovariectomized. Caprine pre-antral follicles in group 1, 2, 3 and 4 were cultured in growth culture medium, growth culture medium + EGF, growth culture medium + IGF-I and growth culture medium + IGF-I + EGF, respectively, for 9 days. The results indicated that EGF (50 mg/l) increased the survival rate of oocytes, but decreased the growth rate of oocytes; IGF-I (100 mg/l) effectively maintained the survival of oocytes and stimulated their growth; IGF-I (100 mg/l) and EGF (50 mg/l) in combination produced a higher effect on both the survival and the growth rate of oocytes than IGF-I or EGF alone. Conclusively, the growth factors can effectively maintain the survival of caprine oocytes from pre-antral follicles and regulate their growth in culture. EGF and IGF-I in association could synergically meliorate the culture system of caprine pre-antral follicles.  相似文献   

Influence of Seasons, Extenders, Slow and Rapid Freezing on Seminal Characters in Korean Native Bucks     

Chang-Yong Choe  Jung-Gon Kim  Sang-Rae Cho  Dong-Soo Son  Young-Keun Kim  S Balasubramanian  Sang-Yong Choe  Gyu-Jin Rho 《Reproduction in domestic animals》2006,41(1):55-60

The success of AI technology is based on both semen quality and freezing process. In order to establish the semen freezing techniques in Korean native bucks, factors affecting the success were evaluated in the present study. Semen collected by electro-ejaculation from bucks during four distinct seasons was evaluated for semen volume and pH, sperm motility and survivability. The semen volume, concentration and total cell were higher in spring, summer and less in winter. Yet, there were no seasonal differences in the proportional data of live sperm, motility score and pH of semen among seasons. The percentage of live sperm after thawing was found to increase with increased concentration of lactose in Tris-Egg yolk-glycerol (TY-G), being highest in TY-G supplemented with 180 mm lactose (TYL180-G), but did not differ between TY-G and TYL120-G. Sperm motility was enhanced by employing 2.0 h equilibration time with rapid freezing method. In conclusion, semen could be frozen with high success rates for further use of AI in breeding techniques and to preserve the Korean native bucks.  相似文献   

Identification of single-nucleotide polymorphisms of the prion protein gene in sika deer (Cervus nippon laiouanus)     

Hyun-Jeong Jeong  Joong-Bok Lee  Seung-Yong Park  Chang-Seon Song  Bo-Sook Kim  Jung-Rae Rho  Mi-Hyun Yoo  Byung-Hoon Jeong  Yong-Sun Kim  In-Soo Choi 《Journal of veterinary science (Suw?n-si, Korea)》2007,8(3):299-301

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Presumed secondary immune‐mediated haemolytic anaemia following elapid snake envenomation and its treatment in four dogs          下载免费PDF全文

HM Ong  A Witham  K Kelers  M Boller 《Australian veterinary journal》2015,93(9):319-326

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Oxygenation and plasma endothelin‐1 concentrations in healthy horses recovering from isoflurane anaesthesia administered with or without pulse‐delivered inhaled nitric oxide     

Tamara Grubb  Anna Edner  Jan HM Frendin  Pia Funkquist  Anneli Rydén  Görel Nyman 《Veterinary anaesthesia and analgesia》2013,40(6):e9-e18

ObjectiveTo assess oxygenation, ventilation‐perfusion (V/Q) matching and plasma endothelin (ET‐1) concentrations in healthy horses recovering from isoflurane anaesthesia administered with or without pulse‐delivered inhaled nitric oxide (iNO).Study DesignProspective experimental trial.AnimalsHealthy adult Standardbred horses.MethodsHorses were anaesthetized with isoflurane in oxygen and placed in lateral recumbency. Six control (C group) horses were anaesthetized without iNO delivery and six horses received pulse‐delivered iNO (NO group). After 2.5 hours of anaesthesia isoflurane and iNO were abruptly discontinued, inhaled oxygen was reduced from 100% to approximately 30%, and the horses were moved to the recovery stall. At intervals during a 30‐minute period following the discontinuation of anaesthesia, arterial and mixed venous blood gas values, shunt fraction (Qs/Qt), plasma ET‐1 concentration, pulse rate and respiratory rate were measured or calculated. Repeated measures anova and a Bonferroni post hoc test was used to analyze data with significance set at p <0.05.ResultsAt all time points in the recovery period, NO horses maintained better arterial oxygenation (oxygen partial pressure: NO 13.2 ± 2.7–11.1 ± 2.7 versus C 6.7 ± 1.1–7.1 ± 1.1 kPa) and better V/Q matching (Qs/Qt NO 0.23 ± 0.05–0.14 ± 0.06 versus C 0.48 ± 0.03–0.32 ± 0.08%) than C horses. Mixed venous oxygenation was higher in NO for 25 minutes following the discontinuation of anaesthesia (NO 6.3 ± 0.2–4.5 ± 0.07 versus C 4.7 ± 0.6–3.7 ± 0.3 kPa). In both groups of horses arterial oxygenation remained fairly stable; venous oxygenation declined over this time period in the NO group but still remained higher than venous oxygen in the C group. ET‐1 concentrations were higher at most time points in C than NO. Changes in other parameters were either minor or absent.Conclusions and Clinical RelevanceDelivery of iNO to healthy horses during anaesthesia results in better arterial and venous oxygenation and V/Q matching (as determined by lower Qs/Qt) and lower ET‐1 concentrations throughout a 30‐minute anaesthetic recovery period.  相似文献   

New Constituents from the Korean Sponge Plakortis simplex     

Jung Soon Oh  Buyng Su Hwang  Ok-Hwa Kang  Dong-Yeul Kwon  Jung-Rae Rho 《Marine drugs》2013,11(11):4407-4418

Six new cyclic peroxides (1–6) were isolated from the Korean sponge Plakortis simplex, along with two new alkylpyridinium alkaloids (7 and 8). The structures of these compounds were completely determined by a combination of NMR analysis and chemical reactions. Compounds 1–6 exhibited cytotoxic/antifungal activities against RAW264.7 cells and Candida albicans.  相似文献   

Characterization of Transgenic Pigs That Express Human Decay Accelerating Factor and Cell Membrane‐tethered Human Tissue Factor Pathway Inhibitor     

HJ Lee  BC Lee  YH Kim  NW Paik  HM Rho 《Reproduction in domestic animals》2011,46(2):325-332

The expression of human complement regulatory proteins (hCRP; hDAF, hCD59, and hMCP) in pig tissues has been suggested as one of strategies to overcome the hyperacute rejection (HAR) in pig‐to‐human transplantation. Expression of human tissue factor pathway inhibitor (hTFPI) in porcine endothelial cells has been suggested as a remedy to overcome microvascular thrombosis. To investigate the effects of these combined transgenes, we established transformed pig cells expressing human decay accelerating factor (hDAF) under the control of enhancer promoter (5′LTR‐PCMVIE), and the fusion protein (hTFPI/hCD4) consisting of the functional domains (K1 and K2) of hTFPI and membrane‐tethering domains (D3 and D4) of hCD4 under the control of PCMVIE. Transgenic pigs were generated with the transformed porcine cells through somatic cell nuclear transfer (SCNT) technology. Analysis of quantitative PCR and real‐time quantitative RT‐PCR showed that four copies of hDAF were integrated and 391 copies of hDAF mRNA expressed in the cells of the transgenic pig. The enhancing activity of 5′LTR was approximately 2 fold compared to CMVIE promoter only. The cell viability test showed that more than 80% of ear cells were viable in the presence of 50% human serum. The chromogenic substrate assay and immunocytochemical staining with tail cells showed that the TFPI activity of fusion protein was observed on the cell membrane. The membrane localization of hDAF and hTFPI proteins was observed by immunocytochemical staining, and the expression of transgenes in heart and liver tissues was also confirmed by immunohistochemistry.  相似文献