In this study, we examined the effectiveness of in vitro fertilization of porcine immature oocytes on the embryo development of blastocysts or hatched blastocysts and the number of cells according to the in vitro fertilization conditions. In the in vitro fertilization of in vitro matured porcine oocytes, there were no significant differences between treatment groups regarding fertilization rate, blastocyst rate, and embryo development of hatched blastocysts according to the storage periods of liquid sperm of 24, 48, and 72 hours. The embryo development rate of hatched blastocysts after the fertilization according to different spermatozoa concentrations (, , and cells/ml) showed the highest rate in the group with a spermatozoa concentration of cells/ml; in particular, this rate was significantly higher than that in the cells/ml group (p<0.05). The total number of blastocysts cells as well as trophectoderms (TE) that developed in each treatment group were also significantly higher in the cells/ml group than in any other groups (p<0.05). In contrast, the embryo development rate of blastocysts according to different co-incubation periods of sperm and oocyte (1, 3, and 6 hr) was high in the 6-hour group; in particular, the rate was significantly higher than that of the I-hour group (p<0.05). Furthermore, the total number of oocytes cells and TEs that developed was significantly higher in the 6-hour group than any other group (p<0.05). In this study, the most effective treatment conditions for porcine embryo development and high cell number were found to be as follows: a sperm storage period of less than 72 hours, a spermatozoa concentration of cells/ml, and a 6-hour co-incubation period for sperm and ooocyte.

The first morphogenetic event of preimplantation development, compaction, was required efficient production of porcine embryos in vitro. Compaction of the porcine embryo, which takes place at post 4-cell stage, is dependent upon the adhesion molecule E-cadherin. The E-cadherin through -catenin contributes to stable cell-cell adhesion. Rho-associated kinase (ROCK) signaling was found to support the integrity of E-cadherin based cell contacts. In this study, we traced the effects of ROCK-1 on early embryonic development and structural integrity of blastocysts in pigs. Then, in order to gain new insights into the process of compaction, we also examined whether ROCK-1 signaling is involved in the regulation of the compaction mediated by E-cadherin of cellular adhesion molecules. As a result, real-time RT-PCR analysis showed that the expression of ROCK-1 mRNA was presented throughout porcine preimplantation stages, but not expressed as consistent levels. Thus, we investigated the blastocyst formation of porcine embryos treated with LPA and Y27632. Blastocysts formation and their qualities in LPA treated group increased significantly compared to those in the Y27632-treated group (p < 0.05). Then, to determine whether ROCK-1 associates embryonic compaction, we explored the effect of activator and/or inhibitor of ROCK-1 on compaction of embryos in pigs. The rate of compacted morula in LPA treated group was increased compared to that in the Y27632-treated group (39.7 vs 12.0%). Furthermore, we investigated the localization and expression pattern of E-cadherin at 4-cell stage porcine embryos in both LPA- and Y27632-treated groups by immunocytochemical analysis and Western blot analysis. The expression of E-cadherin was increased in LPA-treated group compared to that in the Y27632-treated group. The localization of E-cadherin in LPA-treated group was enriched in part of blastomere contacts compared to that Y27632-treated group. ROCK-1 as a crucial mediator of embryo compaction may plays an important role in regulating compaction through E-cadherin of the cell adhesion during the porcine preimplantation embryo. We concluded that ROCK-1 gene may affect the developmental potential of porcine blastocysts through regulating embryonic compaction.

본 논문에서는 저열포틀랜드 시멘트와 steel aggregates인 Ferro-Silicon, 실리카흄, 충전재로서 미세석영과 고강도화에 따른 취성파괴 문제를 개선하기위해 강섬유를 사용하여 압축강도 400MPa이상의 초고강도 분체 콘크리트를 개발 하고자 하였다. 콘크리트의 초고강도화의 영향을 고려하여 물-시멘트비 저감이 가능한 저열포틀랜드 시멘트와 비교대상으로 보통포틀랜드 시멘트를 사용하고, 골재 대체 재료로 Ferro Silicon을 각각의 배합비, 양생조건을 달리하여 압축강도를 비교분석 하였다. 초고강도 콘크리트는 보통콘크리트와 달리 사용재료의 영향이 대단히 중요하며, SEM 촬영결과 Type III, Type IV의 C-S-H수화물이 비교적 많이 생성되었고, 고온고압양생으로 토버모라이트와 조놀라이트가 생성된것을 확인 하였다. 또한 골재의 세립화, 분체의 치밀충전화 및 반응성 재료의 사용으로 인해 페이스트가 고강도화 되고, 강섬유를 사용하여 인성을 보강하므로써, 28일 압축강도 420Mpa의 초고강도 분체콘크리트를 성공적으로 개발 하였다.