• Reproductive and Developmental Biology
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• v.32 no.1
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• pp.9-14
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• 2008

Efficient gene transfer into hematopoietic stem cells is a great tool for gene therapy of hematopoietic disease. Retrovirus have been extensively used for gene delivery and gene therapy. However, current in vitro gene transfer has some obstacles suck as induction of differentiation loss of self-renewal capacity, and down-regulation of homing efficiency for in vitro hematopoietic stem cells transplantation. To overcome these problems, we developed efficient in vitro retroviral transfer technique by direct intra-bone marrow injection (IBM). We identified effective retrovirus gene transfer in bone marrow hematopoietic cells in vitro. Two weeks after retrovirus transfer via IBM injection, we observed stable EGFP gene expression in bone marrow, lymph node, spleen, and liver cells. In addition, $6.4{\pm}2.7%$ of hematopoietic stem/progenitor cells were expressed EGFP transgene from flow cytometry analysis. Our results demonstrate that in vitro retrovirus gene transfer via IBM injection can provide a viable alternative to current or moo gene transfer approach.

• Journal of Animal Science and Technology
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• v.54 no.4
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• pp.283-290
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• 2012

Cryopreservation induces sublethal damage to the spermatozoa, which leads to their reduced fertile life. The objective of this study was to investigate the effect of taurine, hypotaurine and trehalose as antioxidants on the function of the freezing-thawed sperm in Korean Jeju Black Bull. The semen was cryopreserved with tris egg yolk extendercontaining 7% glycerol and treated with 20mM taurine, hypotaurine and trehalose. Frozen-thawed sperms were evaluated for sperm motility, viability, membrane integrity, acrosome integrity and sperm penetration ability. The results were compared to semen cryopreserved in tris egg yolk extender containing 7% glycerol only as control. Frozen-thawed semen evaluation clearlyindicated that the addition of taurine or hypotaurine significantly improved (p<0.05) the motility and viability compared to control spermatozoa. Moreover, in membrane integrity, swollen sperm ratio was significantly increased (p<0.05) in taurine, hypotaurine or trehalose compared to control. In sperm acrosome integrity, F pattern ratio was increased (p<0.05) in hypotaurine among treatments, and AR pattern was significantly lowered (p<0.05) in taurine, hypotaurine and trehalose. In assessed sperm fertilizing ability, taurine, hypotaurine or trehalose significantly improved (p<0.05) the ratio of pronucleus formation and SFI. Finally, compared with the control, addition of taurine, hypotaurine or trehalose as an antioxidant to the freezing extender showed more positive effects on the frozen-thawed spermatozoa. It is concluded that the addition of taurine, hypotaurine, or trehalose to the freezing extender could reduce cryodamage of the Korean Jeju Black Bull spermatozoa.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.6
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• pp.514-520
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• 2019

The reduction in performance due to inbreeding (i.e. inbreeding depression) has long been documented in plant and animal populations. The population of Korean native pigs are small and this breed is valuable in Korea.. This study was aimed to investigate effects of inbreeding depression on total number of piglets born (TNB) and number of piglets born alive (NBA) in Korean native pigs. We used 2,806 pedigree and 303 sows's data with 483 phenotypic records. After estimating genetic parameters for each traits, inbreeding depression was estimated using a mixed model in which the inbreeding coefficient was included as a covariate. Korean native pigs had high heritability for each traits. Inbreeding coefficient constantly increased from 1998 to 2017~2018 but there is no reduction for each traits in 2017~2018 in compared with those in 1998. Significant inbreeding depression was detected for TNB (p=0.03) but not for NBA (p=0.41). In addition there are significantly positive interactions between inbreeding coefficient and breeding value for both traits (p<0.05). These results suggest that Korean native pigs are still having genetic variation for TNB and NBA, which could overcome reproductive risks from inbreeding coefficient increase.

• Journal of Animal Environmental Science
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• v.17 no.1
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• pp.39-48
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• 2011

The present survey was conducted to provide basic information on automatic milking system (AMS) in relation to purchase motive, milk yield and quality, customer satisfaction, difficulties of operation and customer suggestions, etc. Purchase motives of AMS were insufficient labor (44%), planning of dairy experience farm (25%), better performance of high yield cows (19%) and others (6%), respectively. Average cow performance after using AMS was 30.9l/d for milk yield, 3.9% for milk fat, 9,100/ml for bacterial counts. Sixty-eight percentage of respondents were very positive in response to AMS use for their successors but 18% were negative. The AMS operators were owner (44%), successor (44%), wife (6%) and company worker (6%), respectively. The most difficulty (31%) in using AMS was operating the system and complicated program manual. The rate of response to system error and breakdown was 25%. The reasons for culling cow after using AMS were mastitis (28%), reproduction failure (19%), incorrect teat placement (12%), metabolic disease (7%) and others (14%), respectively. Fifty-six percentages of the respondents made AMS maintenance contract and 44% did not. Average annual cost of the maintenance contract was 6,580,000 won. Average score for AMS satisfaction measurement (1 to 5 range) was 3.2 with decrease of labor cost 3.7, company A/S 3.6, increase of milk yield 3.2 and decrease of somatic cell count 2.8, respectively. Suggestions for the higher efficiency in using AMS were selecting cows with correct udder shape and teat placement, proper environment, capital and land, and attitude for continuous observation. Systematic consulting was highly required for AMS companies followed by low cost for AMS setup and systematization of A/S.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.275-289
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• 2002

The objective of this study was to develop an in vitro assessment of sperm fertilizing capacity of bulls and investigate the factors influencing sperm function and characteristics of frozen-thawed bovine spermatozoa. in vitro fertilization (IVF), the evaluation of motility and normal morphology, HOST (hypoosmotic swelling test), Ca-ionophore induced acrosome reaction, luminol and lucigenin-dependent chemiluminescence for the measurement of reactive oxygen species (ROS), the measurement of malondialdehyde formation for the analysis of lipid peroxidation (LPO), and the evaluation of DNA fragmentation using the method of 747-mediated nick end labelling (TUNEL) by flow cytometry were performed in frozen-thawed bovine spermatozoa. Correlations between the rates of fertilization, blastocyst formation after IVF and the values of respective assays were investigated. 1. IVF rate and blastocyst formation rate averaged 64.4% and 34.3% for spermatozoa from high -fertility bull group and averaged 18.5% and 6.2% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. Sperm motility and percentage acrosome reaction averaged 79.0% and 66.2% for spermatozoa from high-fertility bull group and averaged 40.7% and 22.9% for spermatozoa from low-fertility bull group, respectivitely. There were not different between two bull groups. 2. Luminol depenent chemiluminescence, LPO and DNA fragementation averaged 6.4, 2.0 nmol and 2.6% from spermatozoa from high-fertility bull group and averaged 6.5, 3.1 nmol and 7.4% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. There was no significant difference in lucigenin dependent chemiluminescence between two bull groups. 3. Fertilization rate was positively correlated with motility and the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between fertilization rate and the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. 4. Blastocyst formation rate was positively correlated with the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between blastocyst formation rate and motility, the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. In conclusion, these data suggest that ROS significantly impact semen quality. The assays of this study may provide a basis fur improving in vitro assessment of sperm fertilizing capacity.

• Journal of Embryo Transfer
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• v.20 no.2
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• pp.185-190
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• 2005

This study was performed to investigate the effects of multiple superovulation and parity on embryo production in Hanwoo cows. Donors were superovulate 4 times $1\~2$ months interval and inserted CIDR plus (with the capsule of estradiol benzoate 10mg) on Day 10 from standing heat for 9 days and injected 2.5ml FSH (Antorin R-10) 2 times in a day on 6th day to 10th day from insertion of CIDR and the doses of FSH were decreasing 0.5ml on every 2 times. On 3th day of FSH injection, 25ml $PGF_2{\alpha}$ were injected i.m. and on 4th day, CIDR was removed. After 2 days from removing CIDR, AI was performed 2 times 12 hour apart with 2 straws of Korean Proved frozen Semen and simultaneously 200ug/ml GnRH was injected and embryos were recovered on 7th day from Al. The response rates of superovulated donors were $85.7\%,\;90.5\%,\;62.5\%,\;100\%$ from 1 to 4 times of superovulation, respectively. There were significant differences among No. of superovulation times (P<0.05). The results of transferable embryos were 3.7, 3.4, 3.4, 5.7 from 1 to 4 times of superovualtion, respectively. There were no differences among No. of superovulation times. The results of transferable embryos were 2.5, 3.0, 5.3, 3.0, 3.4 form heifer, first born to 4 the born, respectively. There were significant differences among the parities of donors (P<0.05). These results suggested that even 4 times of superovulations of Hanwoo donors could be able to recover transferable embryos, it might be used the donors maximally and improved the adaptation of embryo transfer to farms safely.

• Korean Journal of Weed Science
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• v.3 no.1
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• pp.14-22
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• 1983

To study ecological characteristics of Cyperus serotinus occuring in Korea its propagules were collected from 6 locations from the northern part to the southern part of Korea (Chuncheon, Suweon, Iri, Jeonju, Gwangju, Milyang) in 1981, cultured and replanted 4 times (May 20, June 5, June 20, July 5) in 1982. They flowered from August 10 to August 29 in the plants planted on May 20 and from August 22 to September 4 in the plants planted on July 5. Plant height, number of tillers and top fresh weight were 85-100cm, 375-1,500 tillers/$m^2$ and 500-1,750g/$m^2$, respectively, when they were planted on May 20, and 58-67cm, 300-625 tillers/ $m^2$ and 125-250g/$m^2$, respectively, when they were planted on July S. Weight of seeds and number of rhizomes per plant were 20-50g/$m^2$ and 20.75-61, respectively, whey, they were planted on May 20, and 5-17.5g/$m^2$ and 51.5-80.25 when they were planted on July 5. Local collections showed. the same morphological characteristics at the level of species identification, but there existed variations among the local collections. Cyperus serotinus from Chuncheon and Suweon were longer in the length of inflorescence, than those from Gwangju and Milyang and rhizomes from Chuncheon and Suweon were thicker than the others. Each of local collections may be regarded as different ecotype based on the above mentioned differences in morphology, growth and flowering response to the planting date. The results appear to imply that Cyperus serotinus weeds occuring in various locations of Korea are different one another in competitive ability with rice crop.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.235-243
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• 2002

This study was designed to examine the ability of the bovine (MII) oocytes cytoplasm to support several mitotic cell cycles under the direction of differentiated somatic cell nuclei of bovine, porcine, mouse and human. Bovine GV oocytes were matured in TCM-199 supplemented with 10% FBS. At 20h after IVM, recipient oocytes were stained with 5 $\mu\textrm{g}$/$m\ell$ Hoechst and their 1st polar body (PB) and MII plate were removed by enucleation micropipette under UV filter. Ear skin samples were obtained by biopsy from an adult bovine, porcine, mouse and human and cultured in 10% FBS added DMEM. Individual fibroblast was anlaysed chromosome number to confirm the specificity of species. Nuclear transferred (NT) units were produced by electrofusion of enucleated bovine oocytes with individual fibroblast. The reconstructed embryos were activated in 5 $\mu$M ionomycin for 5 min followed by 1.9 mM 6-dimethylaminopurine (DMAP) in CR1aa for 3 h. And cleaved NT embryos were cultured in CR1aa medium containing 10% FBS on monolayer of bovine cumulus cell for 8 days. Also NT embryo of 4~8 cell stage was analysed chromosome number to confirm the origin of nuclear transferred somatic cell. The rates of fusion between bovine recipient oocytes and bovine, porcine, mouse and human somatic cells were 70.2%, 70.2%, 72.4% and 63.0%, respectively. Also, their cleavage rates were 60.6%, 63.7%, 54.1% and 62.7%, respectively, there were no differences among them. in vitro development rates into morula and blastocyst were 17.5% and 4.3% in NT embryos from bovine and human fibroblasts, respectively. But NT embryos from porcine and mouse fibroblasts were blocked at 16~32-cell stage. The chromosome number in NT embryos from individual fibroblast was the same as chromosome number of individual species. These results show that bovine MII oocytes cytoplasm has the ability to support several mitotic cell cycles directed by newly introduced nuclear DNA.

• Korean Journal of Animal Reproduction
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• v.25 no.1
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• pp.19-28
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• 2001

The present study was undertaken to investigate the effects of cooling rate and equilibration time on the survival, in vitro maturation and development to embryos of frozen-thawed bovine immature oocytes(Germinal Vesicle Stage). The cryoprotectants are used 10% ethylene glycol(EG) as permeating cryoprotectant and 0.05M soc.ose(S) or trehalose(T) as low molecular weight nonpermeating cryoprotectants and 5% ficoll(F) or polyvinylpyrrolidone(PVP) as high molecular weight nonpermeating cryoprotectants. Four freezing solution were uysed in this experiment(EFT: 10% EG + 5% F + 0.05M T, EFS: 10% EG + 5% F + 0.05M S, EPT: 10% EG + 5% P + 0.05M T, EPS: 10% EG + 5% P + 0.05M S). The best equilibration time and freezing solution was 15 min in EPT(83% survival rate of frozen-thawed bovine immature oocytes). When frozen-thawed bovine oocytes were cultured following IVM and IVF, there was no significant difference in cleavage and development rates among the EFT, EFS, EPT and EPS solutions. When 9 blastocysts derived from frozen bovine oocytes were transferred to 6 recipients, two recipients were pregnant. And one was aborted at 45 days of pregnancy and the other had a stillbirth.

• Korean Journal of Animal Reproduction
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• v.27 no.1
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• pp.45-51
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• 2003

This study was conducted to investigate the effects of castration and ovariectomy on growth performance and plasma hormone concentration in pigs. A total of 48 pigs of 35 days of age were used. The results obtained in the present study are summarized as follows: 1. No significant difference was found in average daily gain between ovariectomy group (898.6g) and control gilt group (862.7g), and between castration group (926.0g) and control boar group (945.5g), respectively. Average daily gain of control boar group, however, was significantly higher than that of control gilt group (p<0.05). There was no significant difference in feed/gain between ovariectomy and control gilt group and between castration and control boar group, respectively. Backfat thickness was significantly (p<0.05) higher in ovariectomy or castration group than in control gilt or boar group, respectively. 2. Plasma concentration of IGF-I was significantly (p<0.05) increased during the period of 5 weeks of age (45.1 $\pm$0.72 ng/ml) to 15 weeks of age (356.3$\pm$3.05 ng/ml), and maintained constantly afterwards in control gilt group, as was in control boar group. That of ICF-I tended to be lower in ovariectomy or castration group than in control gilt or boar group, respectively. Regarding steroid hormones of estradiol-17$\beta$, progesterone, and testosterone, the concentration was extremely low at 5 weeks of age, however, increased from 11 weeks to 23 weeks of age in control gilt or boar group, while it was nearly under detection limit in ovariectomy or castration group. 3. Chemical compositions of pork loins were not affected by ovariectomy or castration, except that crude ash content was significantly (p<0.05) higher in castration group than in control boar group. These results indicated that ovariectomy or castration had no effects on growth performance and feed utilization. However, the concentration of sex steroid hormones was under detection limit in ovariectomy and castration group. Further studies, however, are needed to develope the techniques which minimize the stress related with castration or ovariectomy for the production of high quality pork.