Survivin protein expression in bovine follicular oocytes and their in vitro developmental competence

This study examined the relationship between survivin expression and the stage of development of in vitro cultured bovine oocytes and embryos; and whether survivin expression is affected by the quality of cumulus–oocyte complexes (COCS) or the quality of pre-implantation embryos. A polyclonal antibody was prepared using recombinant bovine survivin protein. Expression of survivin mRNA and protein was analyzed by real-time quantitative RT-PCR and immunocytochemistry. In the first experiment, survivin mRNA expression was examined at developmental stages from germinal vesicle (GV) oocyte to blastocyst, it was significantly decreased after fertilization of matured oocytes (P < 0.05), then increased slightly to the 8-cell stage followed by rapid increases at the morula and blastocyst stages (P < 0.05). In the second experiment, the effect of oocyte quality on survivin protein, pro-apoptotic (bax, caspase-3) and anti-apoptotic (survivin, bax inhibitor) mRNA expression was examined. Survivin protein was more strongly expressed in good quality COCS than in poor quality COCS. The expression of the anti-apoptotic genes, survivin and bax inhibitor, was significantly higher (P < 0.05) and that of the pro-apoptotic genes, bax and caspase-3, was significantly lower (P < 0.05) in good compared to poor quality COCS. The developmental competence of good quality COCS (30.4% blastocysts) was significantly better than that of poor quality COCS. In the last experiment also, we confirmed that significantly higher expression of survivin and bax inhibitor genes and significantly lower expression of bax and caspase-3 genes was resulted in good quality blastocysts than in poor quality blastocysts (P < 0.05). It was concluded that the expression of survivin was related to the quality of COCS, their developmental competence and the quality of in vitro produced blastocysts. Consequently, survivin may be a good candidate marker for embryo quality.     

Progress in Microbial Carotenoids Production

Carotenoids are versatile isoprenoids pigments, play a vital role in the cellular system, starting from antioxidant to gene regulation. Carotenoids are widely used in food, nutraceuticals, and cosmetics owing to their vitamin A, antioxidant and anticancer activities. The demand of carotenoids in various sectors has triggered the research to explore a commercially viable and environmentally friendly production. This article presents a short review of progress in carotenoids production from microbial platforms.     

Ge-Jee-Bok-Ryung-Hwan induces apoptosis in human cervical carcinoma HeLa cells--an endoplasmic reticulum stress pathway--

Ge-Jee-Bok-Ryung-Hwan (GJBRH), a commonly used herb formulation in Korea, Japan and China, caused a decrease of viability in HeLa human cervical carcinoma cells. The treatment of GJBRH resulted in genomic DNA fragmentation as well as the increase of Sub-G1 portion in cell cycle analysis. In this study, GFP-Bax over-expression system showed that Bax, pro-apoptotic Bcl-2 family protein, was translocated to mitochondria by the presence of GJBRH. The treatment of BAPTA-AM, permeable endogenous calcium chelator, inhibited GJBRH-induced caspase-3 and -9 activations, the release of cytochrome c and Smac/DIABLO into cytoplasm and the resultant cell death in HeLa human cervical carcinoma cells. The treatment of BAPTA-AM increased the expression of XIAP, which mediates binding to and inhibiting caspases and showed protective effect, in GJBRH-treated cells. GJBRH induced the expression of Glucose Response Protein 78 (GRP 78), a positive ER stress marker protein. However, BAPTA-AM did not interfere with the ER-stress response pathway that triggers the expression of GRP 78. This study showed that GJBRH induces cell death, which occurs downstream of or parallel to this point in the ER-stress pathway linked to apoptosis. In conclusion, GJBRH induces apoptosis in HeLa cells via ER stress-pathway associated mitochondria-dependent apoptosis mechansim.     

Preparation of porcine small intestinal submucosa sponge and their application as a wound dressing in full-thickness skin defect of rat

Small intestinal submucosa (SIS) sponge was prepared by crosslinking with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). The prepared SIS sponges exhibited elastic and soft property on touch and were ease to handle. The SIS sponges have the pore diameter of 100-200 microm and an interconnective porous structure. The SIS sponges exhibited high water absorption ability over 8000%. The water uptake of SIS sponges decreased as SIS concentration used to manufacture SIS sponge increased. In wound healing test, SIS sponge attained uniform adherence to the wound surface. The SIS sponges absorbed higher extent of exudation for wound than that covered with Tegaderm as control. Wound area contracted above 80% at the 21st postoperative day. The SIS sponge treated wound was almost completely covered with a thin layer of epidermis at 4 weeks. In addition, the dermal collagen in the wound regenerated at only SIS sponges treated wounds. The progress of granulous tissue formation was faster in SIS sponges as wound dressing than in Tegaderm. In conclusion, we found that the SIS sponges might be a potential material as a wound dressing.     

Adenoviral delivery of human connexin37 induces endothelial cell death through apoptosis

Gap junction channels formed of connexins directly link the cytoplasm of adjacent cells and have been implicated in intercellular signaling that may regulate the functions of vascular cells. To facilitate connexin manipulation and analysis of their roles in adult endothelial cells, we developed adenoviruses containing the vascular connexins (Cx37, Cx40, and Cx43). We infected cultured human umbilical vein endothelial cells with control or connexin adenoviruses. Connexin expression was verified by immunoblotting and immunofluorescence. Infection with the Cx37 adenovirus (but not control or other connexin adenoviruses) led to a dose-dependent death of the endothelial cells that was partially antagonized by the gap junction blocker alpha-glycyrrhetinic acid and altered the intercellular transfer of Lucifer yellow and neurobiotin. Cell morphology, Annexin V and TUNEL staining, and caspase 3 assays all implicated apoptosis in the cell death. These data suggest that connexin-specific alterations of intercellular communication may modulate endothelial cell growth and death.     

Active ingredients in cade oil that synergize attractiveness of alpha-ionol to male Bactrocera latifrons (Diptera: Tephritidae)

Cade oil, a commercially available essential oil produced by destructive distillation of juniper, Juniperus oxycedrus L., twigs, is known to synergize the attractancy of alpha-ionol to male Bactrocera latifrons (Hendel). Through chemical fractionation and outdoor olfactometer-based bioassays, seven compounds in cade oil were identified that potentially could provide some level of synergism. Tests with sterile laboratory flies showed that four of the seven compounds (eugenol, isoeugenol, 2-methoxy-4-ethylphenol, and 2-methoxy-4-propylphenol), together with a closely related compound not found in cade oil, 2-methoxy-4-methylphenol, are capable of synergizing the attractiveness of alpha-ionol to male B. latifrons under field conditions. The similarity in structures of these five synergistic compounds shows that there is a response to a core 2-methoxyphenol structure, with fly response little affected by some variation in the composition of the side chain on the number 4 carbon. Because identified synergists were structurally similar, only one compound, eugenol, was selected for further field studies. In an 8-wk weathering test, using released sterile flies, traps baited with alpha-ionol + eugenol had catches comparable with catches at traps baited with alpha-ionol + cade oil, with catches generally increased with a higher eugenol loading. For both eugenol and cade oil, catches tended to be better when these synergists were deployed on separate wicks from the alpha-ionol. Eugenol and alpha-ionol, however, were unable to provide attraction comparable with that of cade oil and alpha-ionol in tests with wild fly populations.