In canine, oocytes are ovulated at the GV (germinal vesicle) stage and they have to fulfill maturation phase before reaching metaphase II stage. The efficiency of in vitro maturation is still very low. Therefore, the aim of this study was to investigate the effect of in vitro maturation on nuclear changes of immature canine oocytes recovered from different reproductive stages ovaries and different culture conditions. The oocytes were cultured in TCM-199 with supplement at 5% and for 72 h. The nuclear maturation of canine oocytes was evaluated with Hoechst 33342 stain under fluorescence microscope (Fig. 1). The results of this study detected differences in in vitro maturation rate between oocytes recovered from follicle status and non-follicle status ovaries. However, these differences were not significant as indicated in Table 1 and Fig. 2. In regard to the effect of culture condition with supplements, we did not found significant differences compared with control group (Table 2, Table 3). One of the reasons for this data could be the conditions that ovaries were exposed during slaughtering process or the long distant transportation of the ovaries. Although these data have not shown clearly significant differences results compared with control, furthermore the different reproductive status ovaries was beneficial for maturation of oocytes in vitro and can be a basic part of knowledge to improve in vitro maturation of canine oocytes.

The purpose of this study was to assess follicular viability and competence through in vitro culture of preantral follicles isolated from vitrified mouse whole ovaries. Mouse preantral follicles were enzymatically isolated from vitrified-warmed and fresh ovaries and cultured for 10 days followed by in vitro oocyte maturation. In vitro matured oocytes were fertilized and cultured to the blastocyst stage. Five minutes pre-exposure to vitrification solution of whole ovaries had significantly higher (p<0.05) oocyte survival and maturation rates than between 10 min exposure groups. Oocyte diameter was significantly smaller (p<0.05) in the 5 and 10 min exposure groups (69.4±2.8 and 67.8±3.1) when compared to that of control group (71.7±2.1). There was no statistical significant difference in blastocyst development rates between vitrification group (8.6%) and the fresh control group (12.0%). The mean number of cells per blastocyst was significantly lower (p<0.05) in the vitrification group (41.9±20.2) than in the fresh control group (55.1±22.5). The results show that mouse oocytes within preantral follicles isolated from the vitrified whole ovaries can achieve full maturation, normal fertilization and embryo development.

The purpose of the present study was to investigate the effect of IFN- on prostaglandin synthesis, cyclooxygenase-2 (COX-2) gene expression in vitro and concentration of progesterone (P4) in endometrial cells. Epithelial and stromal cells cultured in vitro were isolated from bovine endometrium and stimulated with increasing doses of IFN- (0, 0.02, 0.2 and 2 ug/ml). Human chorionic gonadotropin (hCG, 1.5 IU/ml) was used as a positive control. Prostaglandin and levels in the culture media were analyzed by enzyme immunoassays and total RNA was extracted from the cells for RT-PCR. P4 concentrations of blood samples were assayed by chemiluminescent immuno assays system. In epithelial cells, COX-2 gene expression was increased in the presence of IFN- (p<0.05), but it was not significantly different in all groups of stromal cells except for 2 ug/ml IFN- group (p<0.05). Although IFN- did not affect and production in epithelial cells, it decreased and production significantly in stromal cells (p<0.05). In vivo experiment, blood concentration of P4 was significantly increased after addition of IFN- (1 ug/ml). The results indicate that PG production was mediated by COX-2 expression in stromal cells but it was not affected in epithelial cells and this suggest that treatment of IFN- could improve the implantation environment of uterine by maintenance of high P4 concentration.

The purpose of this study was to investigate anti-oxidant effects of loach muscle-derived peptides in vitro and in vivo. Loach muscle peptides were prepared by 4 different methods: boiling (B), enzymatic hydrolysis (E), boiling and enzymatic hydrolysis (BE), alkaline and enzymatic hydrolysis (AE). Two different in vitro analyses, DPPH radical scavenging activity and xanthine-xanthine oxidase-induced superoxide radical scavenging activity, were performed. All the four preparations showed concentration-dependent DPPH radical scavenging activity. However, superoxide radical scavenging activity was found only with AE and E preparations. To evaluate in vivo effects, mice were fed with 10% AE-containing diets for 4 weeks before hepatotoxicity induction with CCl4. In serum glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) levels, total antioxidant levels, and relative hepatic weight ratio, no evidence for anti-oxidant effects was found with AE indicating the absence of anti-oxidant effect in the in vivo mouse experiment. It needs to be clarified why anti-oxidant activity of loach protein hydrolysates was not evident in vivo. Furthermore, these results suggest that in vivo evaluation is crucial in demonstrating anti-oxidant activities.

In the present study, acetone extract and sub-fractions of Albizia amara stem bark were evaluated for their free radical potential. The results showed that the crude extract and all the fractions exhibited antioxidant and free radical scavenging activities under different in vitro assays. Among the different fractions, the ethyl acetate fraction exhibited higher DPPH and ABTS radical scavenging activities than the standard quercetin. A. amara stem bark might be valuable source of natural antioxidants that could be used for medicinal and food applications.

거베라의 조직배양과정 중 증식배양단계에서 LEDs 파장이 기내배양묘의 생육에 미치는 영향을 조사하고 자 이 실험을 수행하였다. 절화용 거베라 ‘바네사’의 증식배양 시 발광다이오드(LEDs) 광질에 따른 생육특 성을 보면 주당 신초수가 red와 red(4) + blue(1) LEDs에서 대조구인 형광등에 비해 많았고 far-red에서 는 다소 증가 또는 blue LEDs에서는 줄어드는 경향이 었으나 그 차이는 통계적 유의성이 없었다. 초장은 red LEDs에서 가장 길었으며 주당 엽수는 red(4) + blue(1) LEDs에서 가장 많았고 엽면적은 far-red LEDs를 제외 하고는 비슷한 경향이었다. 주당 생체중과 건물중은 red LEDs에서 가장 무거웠으나 red(4) + blue(1) LEDs, 형광 등과도 큰 차이가 없었다. 증식배양단계에서 가장 중요 한 것은 신초를 확보하는 것으로 다른 생육특성들이 형 광등 대비 큰 차이가 없는 것으로 보아 red LEDs 또 는 red(4) + blue(1) 혼합 LEDs를 광원으로 사용하는 것이 적당하다고 판단된다.