This study was performed to identify the differentially methylated region (DMR) and to examine the mRNA expression of the imprinted H19 gene in day 35 of SCNT pig fetuses. The fetus and placenta at day 35 of gestation fetuses after natural mating (Control) or of cloned pig by somatic cell nuclear transfer (SCNT) were isolated from a uterus. To investigate the mRNA expression and methylation patterns of H19 gene, tissues from fetal liver and placenta including endometrial and extraembryonic tissues were collected. The mRNA expression was evaluated by real-time PCR and methylation pattern was analyzed by bisulfite sequencing method. Bisulfite analyses demonstrated that the differentially methylated region (DMR) was located between -1694 bp to -1338 bp upstream from translation start site of the H19 gene. H19 DMR (-1694 bp to -1338 bp) exhibits a normal mono allelic methylation pattern, and heavily methylated in sperm, but not in oocyte. In contrast to these finding, the analysis of the endometrium and/or extraembryonic tissues from SCNT embryos revealed a complex methylation pattern. The DNA methylation status of DMR Region In porcine H19 gene upstream was hypo methylated in SCNT tissues but hypermethylated in control tissues. Furthermore, the mRNA expression of H19 gene in liver, endometrium, and extraembryonic tissues was significantly higher in SCNT than those of control (p<0.05). These results suggest that the aberrant mRNA expression and the abnormal methylation pattern of imprinted H19 gene might be closely related to the inadequate fetal development of a cloned fetus, contributing to the low efficiency of genomic reprogramming.

희소 한우인 칡소의 정액 동결을 위해서 레시딘을 기본 희석제로 하는 AndroMed와 Tris-egg yolk extender를 사용하여 정자의 생존율과 활력 조사를 위해서 본 연구를 수행하였다. AndroMed 희석제를 사용하였을 때 생존율과 활력은 와 의 결과를 보였다. 그리고 Tris-egg yolk extender의 경우는 각각 와 결과를 보여 생존율에서는 Tris-egg yolk 희석제가 AndroMed 희석제를 사용하였을 때보다 유의적으로(p

The objective of this work was to determine the effect of corpus luteum (CL) grade on pregnancy rate after embryo transfer in Korean cattle and we found that CL development was linked to pregnancy rate. The in vivo derived blastocyst-stage embryos were transferred to 15 recipients synchronized in the estrus cycles. Based on size and palpable characteristics, CLs were categorized into three grade. The grade three CL is not to be identified by rectal palpation. The pregnancy rates tended to increase with the increase in CL size of recipients. In grade one, two, and three, the pregnancy rates were 62.5%, 50.0%, and 0%, respectively. This result suggests that pregnancy rates after embryo transfer might be affected by the CL status of recipients.

As directly imported apparel brands are actively entering the domestic market, studies of the marketing aspects of these imported apparel products and their overall consumption trend have been done. However, studies of whether are not the directly imported apparel products provide Korean consumers with an appropriate fit are not as common. Therefore, this study investigates the satisfaction with and problems associated with the fit along with the recognition of and preference for directly imported US apparel brands. 100 female college students who purchased directly imported US apparel brands participated in a survey. The results of this survey showed that their recognition of, preference for, possession rate of and satisfaction with the directly imported US apparel brands tend to be high. The major purchasing motivations were the excellent ‘design’ and ‘color’ of the products. The subjects reported that they were satisfied with the quality of ‘materials’ and ‘sewing condition’. However, they were not satisfied with the ‘comfort’ or the ‘size’. Their level of satisfaction with the fit of these products differed depending on the specific item. Their level of satisfaction with the fit of T-shirts and knits was high, whereas the level for pants was relatively low. They complained mostly about the sleeve length and garment length of upper-body garments and coats. They were not satisfied with how the pants fit. The areas of dissatisfaction regarding the pants were the waist girth, the crotch length and the pants length.

This study was conducted to investigate the survival rate of frozen-thawed spermatozoa in equine by glycerol concentration and freezing speed. Two stallions (1 Thoroughbred-13 year old and 1 Arab-7 year old) bred in Korea Racing authority was examined for 1 times in a couple of weeks. Semen was collected by condom method standing heated mare and were centrifuged 650 g for 15 min. and isolated the seminal plasma. Thick fraction of semen was diluted EDTA-Lactose-egg yolk diluents to 1:1 and contained in 0.5 ml straw as 6～14×107cells/ml. Final concentrations of glycerol were 3, 5 and 7% in cryopreseved diluents and added 4 times for 2 hours equilibration. For the freezing, equilibrated straws were located 3 or 5 cm above LN2 gas for 5 or 10 min. Survival rates of pre-frozen sperm were 65.0±13.2%, 68.3±10.4%, 66.7±11.5% and post-frozen were 53.3±23.1%, 45.0±15.0%, 50.0±18.0% in 3, 5, 7% glycerol concentration, respectively. There was no difference between glycerol concentrations. Survival rates of frozen-thawed sperm on freezing speed were 36.7±10.4%, 40.0±7.1%, 30.0±13.2% at 3 cm－5 min and 33.3±11.5%, 31.7± 2.9%, 21.7±10.4% at 3 cm－10 min in 3, 5, 7% glycerol concentration, respectively. Survival rates of frozen-thawed sperm on freezing speed were 43.3±15.3%, 32.0±17.9%, 22.3±15.7% at 5cm－5 min and were 47.5±15.0%, 43.3±12.6%, 48.3±15.3% at 5cm－10 min in 3, 5, 7% glycerol concentration, respectively. There were significantly different between groups (p<0.05). These results suggest that glycerol concentration did not affect cryopreservation of stallion semen within 3～7% but freezing speed affects. In our experiment, the best cryopreservation condition was at 5 cm above LN2 gas for 10 min for pre-freezing and 7% of glycerol concentration. These results lead to commercial AI with frozen-thawed stallion semen.

This study was performed to investigate the characteristics within ages and freezing tolerance of spermatozoa in Jindo Dog. Experimental animals were selected 12 herds within 1～8 year’s old and collected semen for 2 times in a week. Collected semen was evaluated whole volume and sperm number with CASA system (SIAS, Medical Supply, Korea). Then seminal plasma were separated and diluted with modified Tris-egg yolk extender and added 4, 6 and 8% glycerol for 4 times to final concentration and equilibrated for 1.5 hrs. Before and after freezing, equilibrated semen were evaluated the survival rates. Total volume of sperm at 1～2 year old group is as 5.2×108 cells/ ml largest and there were no significance among groups. The motility of 1～2 year old group is highest as 90.9% and there were significance among groups. Abnormal sperm showed similar among groups. The survival rate in terms of pre-freezing and post-freezing were decreased all levels of glycerol and reveled 87.0% to 64.5% in 4%, 87.5% to 51.9% in 6% and 73.4% to 29.7% in 8%, there were significant difference among the groups (p<0.05). These results suggest that the optimal sperm-freezing methods in Jindo Dog are utilized with modified Tris egg-yolk extender with 4% glycerol and were improve the reproductive activity by these methods.

The objective of this study was investigate the superovulation treatment and to relate concentrations of blood urea nitrogen(BUN) in Hanwoo donors. Thirty six, at random stages of the estrous cycle, received a CIDR. Four days later, the animals were superovulated with a total of 28AU FSH (Antorin, 2AU=1 ml) administered twice daily in constant doses over 4 days. On the 3th administration of FSH, CIDR was withdrawn and 25 mg PGF2α was administered. Cows were artificially inseminated twice after estrous detection at 12 hr intervals. The cows received 100 μg GnRH at the time of 1st insemination. Embryos were recovered 7 or 8 days after the 1st insemination. Cows with BUN ＜10, 11～18 and ≥19 mg/dl had return of estrus of 34.6, 30.5 and 30.4 days respectively. Return of estrus after superovulation treatment was not significantly lower for cows with blood urea nitrogen (BUN) above 10 mg/dl than for cows with BUN below 10 mg/dl. Cows with BUN ＜10, 11～18 and ≥19 mg/dl had number of transferable embryos of 3.2±1.2, 5.4±1.9 and 4.1±2.1 respectively.

This studies were conducted to investigate the survival rate of frozen-thawed spermatozoa of Jindo Dog by monosaccharide and freezing rates. Experimental animals were prepared 12 males within 1~8 year's old and collected once in a couple of weeks by digital manuplation methods. Collected semen was diluted 1:1 with Tris-egg yolk extender and added 4, 6 or 8% of glycerol and none, 4 mM glucose or 4 mM fructose as cryoprotectant and was equilibrated for 2 hrs in . In monosaccharide groups, the freezing rate was 5 cm-5 min. above . The survival rates without monosaccharide were , , in 4, 6 or 8% glycerol, respectively. In addition of glucose, the survival rates were , , in 4, 6 or 8% glycerol, respectively and in fructose, were , , in 4, 6 or 8% glycerol, respectively. There showed significantly different between glycerol groups and monosaccharides groups (p<0.05). The survival rates of freezing rate in 5 cm-5 min. group was , , and in 10 cm-10 min. group was , , in 4, 6 or 8% glycerol, respectively. There were significantly different between freezing rates (p<0.05). These results suggest that the addition of fructose with 6%-glycerol and slow freezing improve the survival of frozen-thawed sperm in Jindo Dog.

We investigated the cleavage rate and blastocyst yield for each culture condition to enhance tolerance of cryo-preservation of bovine IVF embryo with relatively lower cryo-tolerance compared to in vivo embryo. The cleavage rate and blastocysts yield for CR1aa, IVMD, IVD, CR1aa+10% FBS were 73.2, 69.3, 72.8, 68.5% and 44.1, 30.8, 33.3, 48.0%, respectively. The values did not differ among each treatments without serum. For embryo vitrification, In vivo and In vitro blastocysts were exposed to VS1(10% glycerin, 0.1 M glucose, 0.1 M sucrose, PEG 1%) for 5 min, and VS2 (10% glycerin, 10% EG, 0.2 M glucose, 0.2 M sucrose, PEG 2%) for 5 min and then VS3 (10% glycerin, 30% EG, 0.3 M glucose, 0.3 M sucrose, PEG 3%) for 1 min. The exposed embryos were then loaded into the 0.25 ml plastic straws and then plunged into liquid nitrogen. The straws were held for period of 1 to 2 weeks before thawing. In embryo viability, no differences in blastocyst re-expansion rates were found between in vivo and in vitro embryos. whereas expansion-BL rates was significantly higher for in vivo-derived embryos (72.7%) when compared to in vitro-derived embryos (51.4%), respectively (P<0.05). In conclusion, our results indicate that combined use of CRIaa culture medium with vitrification might enhance tolerance of cryopreservation for bovine IVF embryo production.

The objective of this study was to investigate the effects of abnormal ovarian cycles after superovulation treatment of Hanwoo donors. Thirty six, at random stages of the estrous cycle, received a CIDR. Four days later, the animals were superovulated with a total of 28AU FSH (Antorin, 2AU=1 ml) administered twice daily in constant doses over 4 days. On the 3th administration of FSH, CIDR was withdrawn and 25 mg was administered. Cows were artificially inseminated twice after estrous detection at 12 hr intervals. The cows received GnRH at the time of Ind insemination. Embryos were recovered 7 or 8 days after the 1st insemination. The cows were considered to have resumed ovarian cyclicity on the day of ovulation if followed by regular ovarian cycles. 50.0 percentage of the cows (18/36) had normal resumption of ovarian cyclicity (resumption within 40 days after superovulation), and 50.0% (18/36) had delayed resumption(resumption did not occur until>40 days after superovulation). Delayed resumption Type II (first ovulation did not occur until 40 days after superovulation, i.e. delayed first ovulation 33.3%) were the most common types of delayed resumptions. The mean numbers of total ova from < 10 and 10 of corpora lutea (CL) was 7.3 and 13.9, respectively. The number of transferable embryos differed between < 10 and 10 CL was 4.2 and 5.1, respectively. 11.1 percentage of the cows (4/36) did not resumption their ovarian cyclicity until 60 days after superovulation treatment.

Oxygen consumption has been regarded as a useful indicator for assessment of mammalian embryo quality. This study was performed to investigate whether oxygen consumption reflects morphological grade of in vivo derived bovine blastocyst-stage embryos (blastocyst). The oxygen consumption of in vitro produced blastocyst was compared to its total cell number. In addition, pregnant rate was measured after transplantation of in vivo blastocysts with different oxygen consumption. The quality of blastocyst collected on day 7 after artificial insemination was categorized as grade I and II (G I and G II) based on microscopic observation of the morphology. Oxygen consumption of blastocyst was measured using a scanning electrochemical microscopy (SECM) and total cell number of in vitro blastocyst was enumerated by counting cells stained by propidium iodide. Pregnancy of recipient cow was confirmed with rectal palpation after 60 days of embryo transfer. The oxygen consumptions of G I blastocysts were significantly higher than those of G II blastocysts ( versus , p<0.05). Total cell numbers of in vitro blastocysts were 74.8, 90.7, and 110.2 in the oxygen consumption of below 10.0, 10.0~12.0, and over respectively. Total cell number was significantly increased in embryos with high oxygen consumption (p<0.05). Pregnant rate in recipient cow was 0, 50, and 85.7% in the transplantation of embryo with the oxygen consumption of below 10.0, 10.0~12.0, and over , respectively. These results suggest that measurement of oxygen consumption may help increase the pregnant rate of bovine embryos.