Improvement of canine somatic cell nuclear transfer procedure

The purpose of the present study on canine somatic cell nuclear transfer (SCNT) was to evaluate the effects of fusion strength, type of activation, culture media and site of transfer on developmental potential of SCNT embryos. We also examined the potential of enucleated bovine oocytes to serve as cytoplast recipients of canine somatic cells. Firstly, we evaluated the morphological characteristics of in vivo-matured canine oocytes collected by retrograde flushing of the oviducts 72 h after ovulation. Secondly, the effectiveness of three electrical strengths (1.8, 2.3 and 3.3 kV/cm), used twice for 20 micros, on fusion of canine cytoplasts with somatic cells were compared. Then, we compared: (1) chemical versus electrical activation (a) after parthenogenetic activation or (b) after reconstruction of canine oocytes with somatic cells; (2) culture of resulting intergeneric (IG) embryos in either (a) mSOF or (b) TCM-199. The exposure time to 6-DMAP was standardized by using bovine oocytes reconstructed with canine somatic cells. Bovine oocytes were used for SCNT after a 22 h in vitro maturation interval. The fusion rate was significantly higher in the 3.3 kV/cm group than in the 1.8 and 2.3 kV/cm treatment groups. After parthenogenesis or SCNT with chemical activation, 3.4 and 5.8%, respectively, of the embryos developed to the morula stage, as compared to none of the embryos produced using electrical activation. Later developmental stages (8-16 cells) were transferred to the uterine horn of eight recipients, but no pregnancy was detected. However, IG cloned embryos (bovine cytoplast/canine somatic cell) were capable of in vitro blastocyst development. In vitro developmental competence of IG cloned embryos was improved after exposure to 6-DMAP for 4 h as compared to 0, 2 or 6h exposure, although the increase was not significantly different among culture media. In summary, for production of canine SCNT embryos, we recommend fusion at 3.3 kV/cm, chemical activation, culture in mSOF medium and transfer of presumptive zygotes to the oviduct of recipient animals. The feasibility of IG production of cloned canine embryos using bovine cytoplasts as recipient of canine somatic cells was demonstrated.     

Physiological changes and growth promotion induced in poplar seedlings by the plant growth-promoting rhizobacteria Bacillus subtilis JS

Photosynthetica - This study aimed to determine the effects of plant growth-promoting rhizobacteria Bacillus subtilis JS on the growth and physiological changes of Populus euramericana and Populus...     

Inhibitory role of RhoA on senescence-like growth arrest by a mechanism involving modulation of phosphatase activity

Recently, negative effects of phosphatase in tumorigenesis and metastasis have been suggested in various tumor types. In this study, we showed that RhoA activation modulated phosphatase during senescence-like arrest in human prostate cancer cells. Under senescence-inducing condition, decreased Erk phosphorylation was detected in caRhoA-transfected cells and inactivation of Erk, but not p38, prevented doxorubicin-induced cell senescence. Cells were induced to senescence by inhibition of phosphatase activity (VHR, MKP3, or PP2A) without additional cellular stress. Of interest, caRhoA prevented doxorubicin-induced decrease of phosphatase. Thus, we postulate that RhoA signaling may protect cells against cellular senescence by maintaining phosphatase activity and Erk dephosphorylation.     

Controlled amine functionalization on conducting polypyrrole nanotubes as effective transducers for volatile acetic acid

Pristine (carboxylated) and aminated polypyrrole nanotubes were successfully fabricated using vapor deposition polymerization with a template. In particular, aminated polypyrrole nanotubes were readily synthesized by modifying the nanotube surface with open polyamine chains. Pristine and aminated polymer nanotubes were used as the transducer to acetic acid vapor. Amino-functionalized nanotubes revealed more enhanced sensitivity than the pristine carboxylated nanotubes with the increasing number of amine spacers due to the increased polymer/analyte partition coefficient and mass uptake of the analyte. Moreover, polyamine-functionalized nanotubes presented a reversible and reproducible response to acetic acid up to 40% sensitivity. The aminated polypyrrole nanotubes demonstrated the potential capability to be excellent transducers for volatile fatty acids in disposable sensors.     

D-glucosamine inhibits proliferation of human cancer cells through inhibition of p70S6K

Although D-glucosamine has been reported as an inhibitor of tumor growth both in vivo and in vitro, the mechanism for the anticancer effect of D-glucosamine is still unclear. Since there are several reports suggesting D-glucosamine inhibits protein synthesis, we examined whether D-glucosamine affects p70S6K activity, an important signaling molecule involved in protein translation. In the present study, we found D-glucosamine inhibited the activity of p70S6K and the proliferation of DU145 prostate cancer cells and MDA-MB-231 breast cancer cells. D-glucosamine decreased phosphorylation of p70S6K, and its downstream substrates RPS6, and eIF-4B, but not mTOR and 4EBP1 in DU145 cells, suggesting that D-glucosamine induced inhibition of p70S6K is not through the inhibition of mTOR. In addition, D-glucosamine enhanced the growth inhibitory effects of rapamycin, a specific inhibitor of mTOR. These findings suggest that D-glucosamine can inhibit growth of cancer cells through dephosphorylation of p70S6K.     

黑线姬鼠两亚种(中国东北地区及俄罗斯远东地区的东北亚种和朝鲜半岛的朝鲜亚种) 的线粒体DNA 序列缺少趋异性

为了检测黑线姬鼠两亚种(来自中国东北地区、俄罗斯远东地区的东北亚种和朝鲜半岛的朝鲜亚种)线
粒体DNA 的变异水平并确定朝鲜亚种的分类地位,我们测序分析了两亚种的线粒体DNA 细胞色素b 的部分序列
(1 054 bp)和控制区的部分序列(860 bp),并与基因库中黑线姬鼠相应的单倍型序列进行了比较。可以看出东
北亚种的序列显示出某些分异,可以被分为2 或3 个亚群,所以我们提出需要更多标本的DNA 分析来确定东北
亚种的分类地位。另外,来自韩国的朝鲜亚种的序列,与来自中国东北地区龙江和哈尔滨的东北亚种的两个亚
群相似(1 个亚群是细胞色素b 的两个单倍型,另1 个是控制区的两个单倍型),表明基于线粒体DNA 序列的遗
传多样性与现今基于形态特征对这些姬鼠的分类所得结果是不一致的。因此我们认为来自韩国的朝鲜亚种是一
个只在形态特异上不同于东北亚种的地方亚种,我们建议通过其他DNA 标记来进一步验证其亚种地位。我们还
认为朝鲜半岛不是最近的冰川期黑线姬鼠残遗种的保护区。     

Proteomic analysis of differential protein expression in atherosclerosis

Although recent studies have shown that several pro-inflammatory proteins can be used as biomarkers for atherosclerosis, the mechanism of atherogenesis is unclear and little information is available regarding proteins involved in development of the disease. Atherosclerotic tissue samples were collected from patients in order to identify the proteins involved in atherogenesis. The protein expression profile of atherosclerosis patients was analysed using two-dimensional electrophoresis-based proteomics. Thirty-nine proteins were detected that were differentially expressed in the atherosclerotic aorta compared with the normal aorta. Twenty-seven of these proteins were identified in the MS-FIT database. They are involved in a number of biological processes, including calcium-mediated processes, migration of vascular smooth muscle cells, matrix metalloproteinase activation and regulation of pro-inflammatory cytokines. Confirmation of differential protein expression was performed by Western blot analysis. Potential applications of the results include the identification and characterization of signalling pathways involved in atherogenesis, and further exploration of the role of selected identified proteins in atherosclerosis.     

利用稻瘟霉模型初级筛选抗肿瘤物质研究进展

稻瘟霉(Pyricularia oryzae)活性物质初筛模型是近年来筛选抗肿瘤活性物质的重要模型之一,具有操作便捷、安全经济等优点。为更好地利用该模型,对其原理、方法、应用现状和存在问题进行了综述,同时介绍了利用其他植物病原真菌作为靶标菌建立抗癌药物初筛模型的应用前景,以期对更有效地获得抗肿瘤活性物质起到一定的积极作用。