Factors Affecting the Development of Somatic Cell Nuclear Transfer Embryos in Cattle

Nuclear transfer is a complex multistep procedure that includes oocyte maturation, cell cycle synchronization of donor cells, enucleation, cell fusion, oocyte activation and embryo culture. Therefore, many factors are believed to contribute to the success of embryo development following nuclear transfer. Numerous attempts to improve cloning efficiency have been conducted since the birth of the first sheep by somatic cell nuclear transfer. However, the efficiency of somatic cell cloning has remained low, and applications have been limited. In this review, we discuss some of the factors that affect the developmental ability of somatic cell nuclear transfer embryos in cattle.     

治沟骨干工程台阶式泄水建筑物的消能防冲研究

进行了可用于治沟骨干坝的3种不同坡度的台阶式泄水建筑物的模型试验。结果表明,在跌落水流时,消力池跃前断面单位能量随斜坡变陡和相对坝高Hdam/yc增加而增大,但当Hdam/yc增加到某一值后,逐渐趋于平缓。消力池跃前断面底部压力yp与堰上临界水深yc呈现正比线性相关,yp大小与台阶个数N无关。比较跃前断面压力修正系数λ>1和λ=1时的消能率,当Hdam/yc=10左右时,其消能率相对误差可达7%左右,但当Hdam/yc≥70时,其相对误差则降至1%。试验结果也同时显示出λ与yp/y1同样有着很好的线性关系,其相关系数r=0.99999。     

Glucocorticoid sensitivity depends on expression levels of glucocorticoid receptors in canine neoplastic mast cells

Glucocorticoid (GC) administration with or without other chemotherapeutic reagents is a commonly used option in the treatment of mast cell malignancies. However, the responsiveness of mast cell tumors to GC treatment varies in individuals, and the regulatory mechanisms determining the GC sensitivity of malignant mast cells remain unclear. Since the expression of the GC receptor (GR) has been reported to be associated with GC sensitivity in human neoplastic lymphocytes, we attempted to investigate the relationship between GR levels and GC sensitivity by using neoplastic mast cells derived from canine mast cell tumors (MCTs). To elucidate the regulatory mechanisms involved in GC responsiveness, we analyzed various canine MCT cell lines and tissue samples from dogs with MCT. While the proliferation of canine MCT cells was suppressed by the addition of GC to the culture, we found that MCT cells derived from humans and rodents, as well as canine lymphoma cells, responded poorly to GC. However, there were also some variations in responsiveness to GC treatment among canine MCT cell lines used in this study. Using real-time polymerase chain reaction and Western blot analysis, we elucidated the relationship between GR expression and responsiveness to GC in canine MCT cells. Furthermore, to assess the involvement of GR expression in GC sensitivity in vivo, clinical investigations were conducted on dogs with cutaneous MCT. Written informed consent was obtained from owners, and the affected dogs were treated with prednisolone (0.5-2.0 mg kg(-1)day(-1), administered orally) 1 or 2 weeks prior to the surgical removal of the tumors. Tumor volume was measured according to WHO criteria both before and after prednisolone treatment, and the GC sensitivity of each MCT was determined on the basis of the reduction in tumor volume. Of the 15 dogs with MCT, 11 responded to treatment with prednisolone completely or partially, whereas 4 dogs showed no response. Examination of clinical samples obtained by surgical removal revealed that GR expression levels were significantly lower in GC-resistant MCT tissues than in GC-sensitive MCT tissues. Thus, these results strongly indicate that GR expression may contribute to GC sensitivity in canine MCT.     

Associations between age of gilts at first mating and lifetime performance or culling risk in commercial herds

Age of gilts at first mating (AFM) is a factor associated with reproductive performance of female pigs. The objectives of the present study were to compare AFM and reproductive performance across parity between three herd groups based on a productivity measurement and to determine lifetime performance by AFM and the herd groups. The female data included 38,212 mated gilts entered between 2001 and 2003, and the herd data included mean measurements from 2001 to 2006 in 101 herds. The average female inventory of the 101 herds was 370.2 females. Females were categorized into five groups: AFM 188-208, 209-229, 230-250, 251-271 or 272-365 days. Three herd groups were formed on the basis of the upper and lower 25th percentiles of pigs weaned per mated female over six years: high-, intermediate- and low-performing herds. Multilevel mixed-effects models were performed to analyze comparisons. The AFMs (± SEM) in the high-, intermediate- and low-performing herds were 239.5 ± 0.22, 247.4 ± 0.21 and 256.7 ± 0.35 days, respectively. As the AFM increased from 209-229 to 272-365 days, annualized lifetime pigs born alive (PBA) decreased from 18.2 to 15.3 pigs, and the number of parities at removal decreased from 4.8 to 4.1 (P<0.05). In parity 1, females with an AFM of 209-229 days had fewer PBA, but had a lower culling risk and shorter weaning-to-first mating interval than those with an AFM of 251-271 days (P<0.05). In conclusion, we recommend management practices such as boar exposure to hasten puberty in gilts and decrease AFM.     

Effect of calcitonin on adrenocorticotropic hormone secretion stimulated by corticotropin‐releasing hormone in the hen anterior pituitary

The presence of a receptor for calcitonin (CT) and the effect of chicken CT (cCT) on adrenocorticotropic hormone (ACTH) secretion stimulated by rat/human corticotropin‐releasing hormone (rhCRH) in the hen anterior pituitary were studied. The specific [¹²⁵I]cCT binding component was present in the plasma membrane of hen anterior pituitary and this binding component had properties of a receptor which has binding specificity to cCT, reversibility, saturable binding, high affinity and limited capacity. When anterior pituitary cells were incubated in vitro, cCT increased the maximal secretion of chicken ACTH stimulated by rhCRH. These results suggest that CT may act directly on the anterior pituitary via its receptor binding and enhances the ACTH secretion by CRH.     

Piebald mutation on a C57BL/6J background

The classic piebald mutation in the endothelin receptor type B (Ednrb) gene was found on rolling Nagoya genetic background (PROD-s/s) mice with white coat spotting. To examine whether genetic background influenced the phenotype in the piebald mutant mice, we generated a congenic strain (B6.PROD-s/s), produced by repeated backcrosses to the C57BL/6J (B6) strain. Although B6.PROD-s/s mice showed white coat spotting, 7% of B6.PROD-s/s mice died between 2 and 5 weeks after birth due to megacolon. The PROD-s/s, s/s and Japanese fancy mouse 1 (JF1) strains, which also have piebald mutations on different genetic backgrounds with B6, showed only pigmentation defects without megacolon. In expression analyses, rectums of B6.PROD-s/s with megacolon mice showed ~5% of the level of Ednrb gene expression versus B6 mice. In histological analyses, aganglionosis was detected in the rectum of megacolon animals. The aganglionic rectum was thought to lead to severe constipation and intestinal blockage, resulting in megacolon. We also observed an abnormal intestinal flora, including a marked increase in Bacteroidaceae and Erysipelotrichaceae and a marked decrease in Lactobacillus and Clostridiales, likely inducing endotoxin production and a failure of the mucosal barrier system, leading ultimately to death. These results indicate that the genetic background plays a key role in the development of enteric ganglion neurons, controlled by the Ednrb gene, and that B6 has modifier gene (s) regarding aganglionosis.