We observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. According to the results from each freezing extender, the sperm membrane integrity (HOST: Hypoosmotic Swelling Test) analysis in TCGGD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Glycerol 3%, Dimethylsulpoxide 3.5 M) is 59.8 ± 0.7, TCGSD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Sucrose 0.1 M, Dimethylsulpoxide 3.5 M) is 59.3 ± 0.5 were significantly higher (p < 0.05) among the experimental groups. And MMPs analysis result, we observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. The expression of active MMP-2 was the highest in sperms frozen in TCGSD and TCGD (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Dimethylsulpoxide 3.5 M), Meanwhile, sperms from the TCGGD and TCGED (Tris 250 mM, Citric acid 88 mM, Glucose 47 mM, Ethylene glycol 3%, Dimethylsulpoxide 3.5 M) group showed lower level of active MMP-2 expression. Together, these results indicate that adding glycerol or sucrose to the sperm freezing buffer would not only suppress MMPs expression but also minimize DNA fragmentation, providing a mean to improve the success rate in the in vitro manipulation of rabbit sperms. Therefore, these results suggest that TCGGD or TCGSD extender method for freezing-thawing of rabbit sperm increased the viability after thawing.

The nature of molecular mechanisms governing embryonic cell block is largely unknown, but recent reports have demonstrated that proper execution of programmed cell death is crucial for this process. The main objective of this study is to determine effects of programmed cell death on porcine oocytes development in vitro after parthenogenesis. Among the blastocysts matured in 3MA, MAP1LC3A and ATG5 RNA gene expression level increased in the order of Cyst < 3MA < RP. However, Casp-3 and TNF-r RNA gene expression level decreased in the order of RP < 3MA < Cyst. Expression of mTOR within the RP-cultured blastocyst was the most highly to the inner cell mass, while 3MA-cultured blastocyst showed very lowest expression in inner cell mass. The expression of mTOR showed a pattern opposite to that of MAP1LC3A. That is, its expression was the lowest in Cyst group. When the enzymatic activity of MMP-2 and MMP-9 was assessed in culture, the level of active MMP-9 was higher expression in the medium of each RP treatment group, with the level of another treatment group being relatively higher. Analyses of TIMP-2 and TIMP-3 revealed that their expression was higher in groups that did not receive RP treatment. More specifically, the level of TIMP-2 was not affected by Cyst treatment, while the level of TIMP-3 was higher in 3MA and RP treatment group. There was highly cell division activation efficiency of parthenogenesis on cultured system of RP supplement IVC medium. Therefore, these results suggest that embryo development was significantly increased in conditional culture medium with active autophagy as compared to common cultured condition. Further investigation of this distinction may enable the development of innovative improvements for the production of porcine somatic cell nuclear transfer.

It is known that bones get damaged by accidents and aging. Since the discovery of Bioglass, various kinds of ceramics have been also found to bond to living bone; some of these ceramics are already being clinically used as bone-repairing materials. In the present study, antibacterial calcium silicate gel (Ag-30CaO·70SiO2 gel) was prepared by sol-gel method in order to control the microstructure, which is related to the dissolution rate and induction period of apatite formation in body environment. In addition, biological Ag-30CaO·70SiO2 is tested. This was done to impart antimicrobial activity to the 30CaO·70SiO2. Ag ion was added during sol-gel synthesis to replace the H2O added during the making of the 30CaO·70SiO2 gel, which has silver solutions of various concentration. After the sol-gel process, 1N-HNO3 solution was used to wash the gel when synthesizing the gel, in order to maintain the porous structure and remove PEG, water soluble polymers. Then, the apatite forming ability of the sol-gel derived CaO-SiO2 gels was investigated using simulated body fluid (SBF), which had almost the same ion concentration as that of human blood plasma. The gels were analyzed by FT-IR spectroscopy, SEM observation, XRD, and fluorescent microscopy. The apatite was successfully created even after washing the gel; apatite is present in an amorphous state, and was found to affect the concentration of the Ag ion in cells in MC3T3 live & dead assay results. From these results, it is suggested that a good material that can be used to repair defects of nature bone is Ag-30CaO·70SiO2 gel.

롤 투 플레이트 시스템은 인쇄 전자에서 가장 경제적이고 효과적인 방법 중 하나이다. 본 논문에서는 롤 투 플레이트 시스템의 틈새출구에서 PDMS의 공동 변화가 유체 흐름의 어떠한 영향을 미치는지, 또 플레이트에 어떻게 전이되는가를 시뮬레이션을 통해 규명하였다. 실험은 IGT-C1 인쇄적성 실험기를 이용하여 전이율, 인쇄 밀도, 프린트 스루, 분할점 등과 같은 데이터를 측정하여, 컴퓨터 시뮬레이션 결과 값들과 비교 분석하였다. 컴퓨터 시뮬레이션은 Flow 모델을 뉴토니언 모델로 생성하여, Navier-Stokes 방정식을 기반으로 한 패키지 소프트웨어인 Polyflow와 Flow 3D로 시뮬레이션 하였고, 시뮬레이션 결과와 실험 결과는 잘 일치하였다.

Roll to roll system을 기반으로 한 인쇄 전자는 유기 박막 트렌지스터(OTFTs), 디스플레이 그리고 센서와 같은 장치를 제작하는데 유용하다. 그 중에서 그라비어 인쇄는 인쇄 전자에 가장 유망한 고속도 roll to roll 기술로서 자리매김하고 있다. 본 연구에서, 우리는 전통적으로 그라비어 인쇄에 사용되어 왔던 저점도 잉크의 단점, 즉 낮은 패턴 충실도와 전도성을 개선하고자 trench 패턴을 기반으로 한 그라비어 연구를 수행하였다. 특히, 미세패턴 형성을 위해 새로운 나노 입자의 실버 페이스트 잉크를 제작하였고 고점도 잉크가 trench로 잘 채워지지 않는 단점을 극복하기 위한 flooding 기술을 소개하였다. 또한, 이러한 환경에 맟는 doctoring 및 printing 조건들을 최적화 시키는 연구를 수행함으로서 printing reliability를 높이고자 하였다. 그 결과 line width 20 ㎛, 두께 2 ㎛ 그리고 resistivity가 1.3E-05 Ωcm인 균일한 패턴을 인쇄할 수 있었다.

The main purpose of this study is to estimate the effect of adding Tea-N-Tris (TES) to the freezing buffer for miniature pig sperm. In particular, we attempted to identify the association between the MMPs expression and the fertility and viability of frozen sperm from each extender (LEY (Lactose Egg-Yolk), TLE (TES + LEY), TFGE (TES + Fructose + Glucose Egg-Yolk)). In accordance with this, Hypoosmotic Swelling Test (HOST) respond test was the lowest among sperms frozen in LEY while the highest HOST respond was observed among sperms frozen in TLE. Furthermore, we observed MMPs expression in all sperm groups, with pro-MMP showing lower expression than active MMPs. The expression of MMP-9 and MMP-2 was the highest in sperms frozen in LEY, Meanwhile, sperms from the TFGE and TLE group showed lower level of MMP-9 and MMP-2 expression in the order of TLE being the lowest. LEY group showed lower rate of blastocyst development than the TES supplement group, although the difference was not statistically significant. Meanwhile the rate of blastocyst development appeared similar when sperms from TLE and TFGE group were used for IVF. Together, these results indicate that adding Tea-N-Tris to the sperm freezing buffer only suppresses MMPs protein activation but also maximize in-vitro fertility, providing a means to improve the success rate in the in vitro manipulation of miniature pig sperm.

To describe the macroscopic anatomy and ovarian-physiological difference of the genital organs of the female Korean water deer, organs from captured animals in a wild area of Korea were dissected. The ovary of estrus group was about 1.10 ± 0.02 mm long and weighed about 0.50 ± 0.02 g. And pregnant group was about 1.3 ± 0.10 mm long and weighed about 0.40 ± 0.05 g. And the crowns of corpora lutea were found in the estrus group, but we couldn't find crowns at the pregnant group. Especially, the estrus ovaries tended (p=0.04) to be heavier than the ovaries during pregnancy. The MMP-9 activity was higher at the Graafian follicles of pregnant group than that in estrus group. However, with regard to follicles of estrus group, MMP-2 level was higher than that in pregnant group. Furthermore, apoptosis detection marker (Casp-3) was highly expressed in Graafian follicle of the pregnant group and the corpora lutea of estrus group. Thus, the differential expression of MMPs observed in this study suggests that the reflected the mechanisms underlying of monovulatory in estrus and/or pregnancy. Our results may be very useful as it provides with information that may be considered for the development of reproductive biotechnologies in endangered animals.

Ink-jet printing technology has been widely attractive due to its facility for direct and fine printing on various substrates. Recent studies have focused on expanding the application of ink-jet printing technology from general consumer use and design companies to the prototype production of precision parts and parts manufacturing. The use of ink-jet printing technology in decorated tableware, tiles, and other ceramic products also has many advantages. The printing process is fast and can be adaptable to various kinds of objects because there is no direct contact point between the printer and the substrates to be printed. For application to ceramic product decoration, inks containing highly dispersed inorganic nano-pigments are required. Here we report the synthesis and characterization of blue CoAl2O4 nanopigment for ink-jet printing. Blue ceramic ink based on the obtained CoAl2O4 pigment was prepared by dissolving CoAl2O4 pigment in a mixed solution of ethylene glycol and ethanol with volume ratios of 7:3 and 8:2, respectively, to obtain the appropriate viscosity for ink-jet printing. The ink solution contained 15 wt% of CoAl2O4 pigment and Cetyltrimethyl ammonium bromide(CTAB) and Sodium dodecyl sulfate(SDS) as dispersive agents. The prepared blue ceramic ink was stably jetted and formed a sphere-shaped droplet from an ink-jet printer.

Bovine coat color is decided by the melanocortin receptor 1 (MC1R) genotype mutation and melanogenesis. Specially, in the various cattle breeds, dominant black coat color is expressed by dominant genotype of ED, red or brown is expressed in the frame shift mutation of recessive homozygous e by base pair deletion and wild type of E+ is expressed in various coat colors. However, not very well known about the effected of MC1R genotype mutation on the coat color through family lines in KBC. Therefore, this study were to investigate effect of MC1R genotype mutation on the coat color, and to suggest mating breed system in accordance with of MC1R genotype for increased on brindle coat color appearance. Parents (sire 2 heads and dam 3 heads) and offspring (total : 54 heads) from crossbreeding in KBC family line with the MC1R genotype and phenotype records were selected as experimental animals. The relationship between melanocortin 1 receptor (MC1R) genotypes expression verified by PCR-RFLP, and brindle coat color appearance to the family line of the cross mating breed from MC1R genotype pattern was determined. As a result, 4MC1R genetic variations, E+/E+ (sire 1), E+/e (sire 2 and dam 3), E+/e with 4 bands of 174, 207 and 328 bp (dam 1) and E+/e with 3 bands of 174, 207, 328 and 535 bp (dam 2) from parents (sire and dam) of KBC. However, 3 genetic variations, e/e (24%), E+/E+ (22%) and E+/e (56%) were identified in offspring. Also, brindle coat color expressrated was the e/e with the 0%, E+/E+ with 67% and E+/e with 77% from MC1R genotype in offspring on the cross mating of KBC. Furthermore, when the sire had E+/e genotype and the dam had E+/E+ with the 3 bands or E+/e genotype, and both had whole body-brindle coat color, 62% of the offspring had whole body-brindle coat color. Therefore, the seresults, the mating system from MC1R genotype patterns of the sires (E+/e) and dams (E+/E+ with the 3 bands or E+/e) with brindle coat color may have the highest whole body-brindle coat color expression in their offspring.

The present study was conducted to compare on embryo survival rates by blastomere isolation methods, and establish the optimal PCR procedure for perform the sexing of bovine blastocysts produced by IVF. IVF embryos used in the study was used the Bisected or Sliced methods for blastomere isolation, and the survival rates of blastocyst with rapid way of sexing PCR was assessed. In the present study for survival rates in blastocyst was the total cleavage rate was 75% and a blastocyst development among cleaved embryos was 40%. Survival rate of embryos treated with intact, bisected or sliced method was 100, 63.3 or 81.3%, respectively. Therefore, survival rate of embryos treated with sliced method was higher compared to that of embryos treated with bisected method. The sexing rate of female or male was not significantly different between S4BFBR primer and BSY + BSP primer (1.75 : 1 vs. 1.43 : 1), respectively. Because of the PCR amplification using the S4BFBR primer was simpler method than multiplex PCR amplification method. Furthermore, the accuracy of sexing rate and reduction of PCR work time between 2-step and 3-step of PCR methods was 98.0% / 1.5 hr and 97.0% / 3.5 hr, respectively. Based on these results, it can be suggested that the sliced and PCR methods we developed was very effective method to reduce time consuming and procedure of PCR amplification for sexing with the increase of survival rate on the blastocyst.

The objective of this study is to estimate the effect of adding TES to LEY and FGE freezing extender for the sperm viability, acrosomal morphology and DNA fragmentation from miniature pig sperm, we evaluated sperm characteristics in TFGE, TLE and LEY with various thawing condition ( for 20 sec, 45 sec and for 5 sec, respectively), and in different concentration of glycerol at 1%, 1.5%, 3%. The sperm viability and normal acrosome intact(NAI) in TFGE (Viability : , NAI : ), TLE (, ) extender significantly(p<0.05) increased than that in LEY (, ) extender thawed at for 5 sec. According to the results from glycerol concentration, the viability and NAI of miniature pig sperm in 1.5% glycerol TLE (, ) was highest among the experimental groups. In accordance with this, DNA fragmentation rates was the lowest in TLE () while that in LEY () is the highest. Therefore, these results suggest that TLE extender method for freezing- thawing of miniature pig sperm increased the viability after thawing.

쇠고기의 육질 등급을 좌우하는 요인으론 육색, 지방색, 조직감 및 상강도(마블링, 근내 지방도) 등이 있는데 그중 근육 내 지방 축적도는 고기의 연도 및 다즙성에 영향을 미치 며 등급을 결정하는 중요한 요인 중 하나이다. 최근 들어 근육 내 지방 합성에 관련된 유전자 연구 및 SNP 탐색을 통한 DNA maker 개발 등 분자생물학적으로 많은 연구보고 가 있으나 등급 간 유의성에 대한 정확한 증명에 관한 연구는 미비하다. 따라서 본 연구는 한우등급 판정소에서 공인하여 선정된 1++의 등급을 판정받은 한우 6두와 2등급으로 판정받은 한우 6두에서 근내지방도와 관련된 유전자로 알려져 있는 GPAT, ACSS2, MGAT, EXT1, FABP4, BCO2, LPL, DGAT, PLIN2, ADSF, TG와 연도와 관련된 유전자인 CAST와 CAPN3의 mRNA 발현양상을 분석하고 유의적 차이점을 확인 하여 경제 형질 선발 마커의 사용 가능성을 선발하기 위하여 등심조직 내 mRNA를 추출 하였고, 각 유전자를 탐침으로 이용하여 sqRT-PCR 및 양적 분석을 위한 nano-spector mate fluorescence system으로 발현양상을 측정하였다. 이후 유의적 평가를 검증 위하여 SAS program (V9.1)의 t-test 검정을 실시하였다. 각 경제 형질 유용 유전자의 발현양상을 분석한 결과 ADSF와 LPL의 경우 평균 근내 지방도가 8±1인 1++등급보다 2±1인 2등급에서 높은 발현양상을 보이고 있었으며, 그 값 이 유의적 차이를 보이지 않았다. 이 외 근내지방도 및 연도와 관련된 유전자 그룹의 경 우 1++등급이 2등급보다 발현양상이 높게 측정되었다. 근내지방도 및 연도에 관련된 유 전자로 선발한 GPAT, ACSS2, DGAT, PLIN2의 경우 유의성 분석결과 유의적 차이점이 없는 것으로 나타났으며, MGAT, EXT1, CAPN3, FABP4, BCO2, CAST, TG의 경우 유의 성(p<0.05)이 있었다. 따라서 본 연구 결과에 의하여 근내지방도 선발 표지 마커로 사용이 가능한 MGAT, EXT1, TG, FABP4, BCO2와 연도 측정에 관련한 CAST, CAPN3의 총 7개의 유전자를 선발 할 수 있었다.

칡소의 고유유전자표현형 방식은 E+/E+와 E+/e 2개의 유전자형만이 출현하고, e/e 유 전자형을 가지는 개체는 전혀 출현하지 않아 제주재래흑우에서와 같이 흑색 호반모가 발 현되기 위해서는 기본적으로 E+ allele이 필요한 것으로 보고됨에 따라 본 연구는 수정란 이식 기법을 이용한 울릉칡소 특화단지 조성 연구용역에 의해 생산되어진 칡소의 모색발 현의 현황에 대해 조사하여 칡소의 우량유전자원의 선발과 증식을 통하여 칡소의 고정과 복원을 위해 실시되었다. 조사에 공시한 칡소는 2007년 수정란 이식 기법을 이용한 칡소 울릉특화단지 조성 연 구용역에 의해서 생산되어진 칡소를 대상으로 실시하였으며, 수정란 이식 후 임신 5-6개 월에 울릉관내 농가에 입식된 한우에서 생산된 칡소(이하 입식우) 159두와 울릉도 관내 의 한우에 칡소 수정란을 이식하여 생산된 칡소(이하 관내이식) 117두를 조사 대상으로 공시하였다. 먼저 울릉칡소의 모색 분포는 황모 24.3%(67/276두), 흑모 13.0%(36/276두) 와 호반모 62.7%(173/276두)의 분포를 보였으며, 입식우의 호반모 출현율이 66%(105/ 159두)로 관내이식에 의해 생산된 칡소의 호반모 비율 58.1%(68/117두)보다 다소 높은 경향이었으며, 성별에 따른 모색의 분포를 조사한 결과 수컷의 경우 황모의 비율이 18.1 %(25/ 138두)로 암컷의, 31.4%(43/137두)에 비해 출현율이 낮은 반면 호반모의 비율은 68.1% (94/138두)로 암컷의 56.9%(78/137두)에 비해 다소 높게 나타났다. 을릉도 칡소 모색 분포에 따른 조사를 바탕으로 비경의 흑반점의 강약에 따른 호반모 의 발현 양상을 분석한 결과 비경에 흑반점이 강한 개체들 중 호반모의 발현 정도를 분 석한 결과 호반모 1, 호반모 2, 호반모 3, 호반모 4, 호반모 5의 비율이 각각 2.3% (3/ 109), 24.8%(27/109), 43.1%(47/109두), 19.3%(21/109두), 10.1%(11/109두)로 나타났으며 흑반점이 중인 개체는 각각 7.9%(3/38), 52.6%(20/38), 34.2%(13/38두), 5.3%(2/38두), 0 %(0/38두)로 나타났다. 또한, 흑반점이 약한 개체는 각각 42.5%(17/40두), 45%(18/40두), 7.5%(3/40두), 0%(0/40두), 5%(2/40두)로 나타났다. 이 결과로 미루어 볼 때 비경에 있는 흑반점의 강약이 호반모 발현에서 황모와 흑모의 비율에 영향을 미치는 것으로 판단된다.

MMP-2, 9의 경우 난포의 발달, 난자의 성장, 그리고 배란 시 extracellular matrix를 재 구성하는 중요한 역할을 하는 것으로 알려져 있으며, 혈청배지 및 무혈청배지를 이용한 체외수정란의 배아발달 과정에 따른 MMPs(2, 9)와 TIMPs(2, 3)의 발현양상의 차이가 있 을 것으로 사료된다. 따라서 본 연구는 소의 체외성숙난자와 체외수정방법에 따른 체외수정란의 발달율과 cumulus cell(CC), single oocyte cell(SOC), cumulus oocyte complex(COC) 및 blastocyst 에서 MMPs와 TIMPs의 활성 및 단백질발현양상을 zymography와 ELISA에 의하여 분석 하였으며, immunofluorescence를 이용하여 발현위치분석을 실시하였다. 체외성숙 이후 Real-time PCR을 이용하여 mRNA의 발현양상을 분석한 결과 MMP-2, 9은 CC가 SOC보 다 높은 발현을 보였으며, TIMP-2, 3의 경우 상반된 발현양상을 보이고 있었다. MMPs의 활성도 분석결과도 mRNA의 결과와 같았으며, 배양배지의 MMPs의 단백질양적분석에서 도 같은 양상을 나타내고 있었으나, TIMP-2의 경우 SOC가 높은 발현을 보였으며, TIMP-3는 CC에서 높은 발현을 나타냈다. 위의 결과를 토대로 COC의 MMPs와 TIMPs의 단백질 작용위치를 분석한 결과 MMP-2는 난자의 세포질을 중심으로 발현양상을 나타내 고 있었으며, MMP-9에 비하여 높은 발현을 확인할 수 있었다. TIMPs의 경우 난자의 세 포질보다 난구세포에서의 발현이 높게 나타났으며, TIMP-3의 발현이 높게 나타났다. 체 외배양방법에 따른 blastocyst의 발달율은 무혈청배지(ES)(61.22% 60/98)가 혈청배지(CR- 1aa)(48.28% 28/58)보다 높은 발달율을 보였다. 체외수정배양배지의 MMPs의 단백질양적 분석결과 MMP-2는 ES가 CR-1aa보다 상대적으로 높은 발현을 보였고, MMP-9은 CR- 1aa가 ES보다 높은 발현을 보였다. TIMPs의 발현양상의 경우 대체적으로 TIMP-3의 발 현이 높게 나타났으며, apoptosis의 활성도 분석에서 Casp-3의 발현이 CR-1aa배양에서 높게 나타남을 확인할 수 있었다. MMPs와 TIMPs의 단백질작용위치를 분석한 결과는 ES는 MMPs와 TIMPs의 발현이 inner cell mass를 중심으로 발현이 높게 나타났으며, trophoblast에서는 낮은 발현이 나타났다. 이와 반대로 CR-1aa의 경우 ES와 다른 위치의 발현을 나타냈으며, TIMPs의 발현은 대체적으로 낮게 발현되었다. 따라서 본 연구 결과 는 무혈청배양 방법이 수정란의 발달 시 MMPs의 활성을 높여 배아형성에 영향을 줄 수 있을 것이라 사료된다.

This study was performed to the expressions of pregnancy-associated plasma protein-A (PAPP-A) and 20alpha-hydroxysteroid dehydrogenase (-HSD) in bovine corpus luteum during early pregnancy. To determine the function of PAPP-A gene during early pregnancy, we collected corpus luteum samples on 30, 60 and 90 days of pregnancy in bovine. The mRNA expression of PAPP-A, -HSD, progesterone-receptor (PR) and insulin-like growth factor binding protein4 (IGFBP4) gene was conducted by Real-time PCR. In parallel with mRNA levels, The protein expressions of PAPP-A and -HSD were detected by immunological analysis. The mRNA expressions -HSD and PAPP-A significantly increased on day 90 in the corpus luteum during pregnancy. The mRNA expression of PR and JGFBP4 in the corpus luteum progressively was enhanced at 30 to 60 day, but decreased on 90 day of pregnancy in the corpus luteum. The expression patterns of these genes, PAPP-A and -HSD were similar pattern in these tissues. In conclusion, PAPP-A and -HSD activity in corpus luteum could be played a role for early pregnancy manifestation.

This study was performed to comprehend the plasma proteins expressed specifically during early pregnancy in pregnant or non-pregnant Hanwoo using proteomic analysis technique. Plasma samples (0, 2, 3, 4, 7, and 11 weeks after AI) were obtained from pregnant (P, n=3) or non-pregnant (NP, n=4) Hanwoo, respectively. To evaluate proteins differentially expressed, 2-dimensional electrophoresis (2DE) was conducted. Normalized protein spots were selected for the significant expression variation deviated over two fold in its expression level between two groups. Molecular functions of the proteins were DNA binding, protein binding, hemoglobin binding, ferrochelatase and transporter activity and arylestera, respectively. According to western blotting, haptoglobin was specifically expressed only in NP group during early pregnancy; however, paraoxonase 1 was highly expressed in pregnant group. Based on these results, pregnancy was maintained successfully by the activation of specific plasma proteins associated with immune system and antioxidant regulation during early pregnancy in Hanwoo

Matrix metalloproteinases (MMP) play important roles in extracellular matrix (ECM) remodeling during ovarian follicular development, oocytes development and ovulation. In an attempt to investigate the effect of MMP activation in development cumulus-oocytes complexes, we examined the localization and expression of MMP, and monitored MMP expression profile. Cumulus-oocytes complexes were collected and matured in vitro for 24 hr, 36 hr and 48 hr. A mRNA expression of MMP-2, MMP-9, TIMP-2 and TIMP-3 was detected in all culture medium regardless of CC, OC and COCs. Activity of MMP-2 in the OC progressively was increased from 24 hr to 48 hr. But MMP-9 was not detected in all culture medium. The localization of MMP-2 was also measured by immunohistochemistry analysis. The MMP-2 and TIMP-2 was detected in cumulus cell and oocyte zone pellucida. Expression of MMP-2 protein in the COCs was progressively increased from 24 hr to 48 hr. However, MMP-9 protein was progressively decreased from 24 hr to 48 hr. And TIMP-2 protein was most highly expressed in the COCs 36 hr. Expression of TIMP-3 protein in the COCs was progressively increased from 24 hr to 48 hr. In conclusion, these results suggest that MMP-2 plays a role in maintaining normal maturation and development by controlling the ECM inhibitor concentration on cumulus cell and oocytes.

This study was conducted to investigate the specific expression genes in the cloned bovine tissues. Donor cells, cloned tissues were analysed by RAPD-RFLP method. The results were detected three genes (CH-U7B, CH-U7M and CH-U7P) in the cloned fetus. It was found a single copy genes by southern hybridization. Sequence analysis of CH-U7M gene was shown 99% homology to a previously reported EST from a cloned bovine fetus. The putative ORF was encode a protein of hydrophobicity index 0.03. Semi-quantitative RT-PCR by using the CH-LS001 specific primer was remarkably detected in the lung tissue of cloned fetus. Further investigation of these genes may provide one of the key information to explain the early death, abnormal fetus, large off-spring and the low pregnancy rate in the production of cloned bovine.

Non-sintering cement was manufactured with briquette ash. Alkali activator for compression bodies used a NaOH solution. In order to apply alkali-activated briquette ash and the non-sintering cement to concrete, several experimental studies were performed. It was necessary to study the binder obtained by means of a substitute for the cement. This study concentrated on strength development according to the concentration of NaOH solution, the curing temperature, and the curing time. The highest compressive strength of compression bodies appeared as 353kgf/cm2 cured at 80˚C for 28 days. This result indicates that a higher curing temperature is needed to get a higher strength body. Also, geopolymerization was examined by SEM and XRD analysis after the curing of compression bodies. According to SEM and XRD, the main reaction product in the alkali activated briquette ash is aluminosilicate crystal.