• Journal of Embryo Transfer
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• v.15 no.1
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• pp.33-38
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• 2000

This study was undertaken in an effort to select the sire bull in Hanwoo through in vitro fertilization of proven bull semen. It was used for in vitro fertilization that of the 20 proven bull semen with follicular oocytes derived from slaughterhouse ovaries of Hanwoo. The stage of maturation on the time course of bovine cumulus-enclosed oocytes incubated for 24 hours was found the highest(96.4%) than hose of other maturationi time. In vitro fertilization rate of bovine oocytes with proven bull sperm showed from 61.5 to 88.9%. Polyspermy of in vitro fertilized oocytes according to proven bulls were the highest KP 491(61.5%) nothing but KP 486, KP 491 and KP 497.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.6
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• pp.1227-1236
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• 2001

Genotoxicological safety on 10 kGy-gamma irradiated vegetable juices such as Oenanthstolonifera DC., Daucus carota L., Brassica oleracea var. acephala and Angelica keiskei was determined by the Salmonella typhmurium reversion assay, the SOS Chromotest using in Escherichia cloi PQ37 and chromosome aberration test in cultured Chinese hamster lung (CHL) fibroblast cells. Vegetable juices exposed to 10 kGy-gamma ray revealed negative results in these three in vitro mutagenetic tests.

• BMB Reports
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• v.36 no.4
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• pp.390-398
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• 2003

The CDP/Cux transcription factor was previously shown to be proteolytically processed at the G1/S transition. In view of characterizing and eventually identifying the protease responsible for CDP/Cux processing, we have established an in vitro proteolytic processing assay. CDP/Cux recombinant proteins expressed in mammalian or bacterial cells were efficiently processed in vitro using as a source of protease either whole cell extracts, the nuclear or the cytoplasmic fraction. Processing was found to take place optimally at a lower pH, to be insensitive to variations in salt concentration, and to be inhibited by the protease inhibitors MG132 and E64D. Interestingly, the bacterially-produced substrate was more efficiently processed than the substrate purified from mammalian cells. Moreover, processing in vitro was more efficient when CDP/Cux substrates were purified from populations of cells enriched in the S phase than in the G1 phase of the cell cycle. Altogether, these results suggest that post-translational modifications of CDP/Cux in mammalian cells inhibits processing and contributes to the cell cycle-dependent regulation of processing. The in vitro processing assay described in this study will provide a useful tool for the purification and identification of the protease responsible for the processing of CDP/Cux.

• Reproductive and Developmental Biology
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• v.30 no.4
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• pp.301-305
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• 2006

The aim of this study was to investigate whether addition of porcine epididymal fluid (pEF) into culture medium during in vitro maturation influences the nuclear maturation of porcine germinal vesicle (GV) oocytes. Porcine cumulus-oocyte complexes (COCs) from follicles were cultured in tissue culture medium 199 (TCM 199) containing pEF. After 48hr of culture, oocytes were examined for evidence of GV breakdown, metaphase I, anaphase-telophase I, and metaphase II. The proportion of oocytes reaching at metaphase II (M II) stage was significantly (p<0.05) increased in oocytes cultured in the media supplemented with 10% pEF during in vitro maturation than in those without pEF regardless of cumulus presence or absence (54.6% vs 22.5%, 51.7% vs 24.2%). The supplementation of pEF during maturation of oocyte enhanced oocytes maturation in a dose-dependent manner in vitro. Also significant differences (p<0.05) in the percentage of MII oocytes were observed according to exposure period in pEF. Present study suggests that pEF contains a enhancing component(s) for nuclear maturation of porcine immature oocytes in vitro.

• Korean Journal of Animal Reproduction
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• v.20 no.3
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• pp.299-305
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• 1996

This study was conducted to investigate the effects of co-culture for the development rate to morula/blastocyst stages of early porcine embryos, derived from oocytes matured and fertilized in vitro, with porcine oviductal epithelial cell monolayers(POEC) in the two different media, respectively. The rates of embryos developed to 2-, 4-, 8∼16-cell and morula/blastocyst stage were 57.2, 48.2, 37.2 and 19.3% in Ham's F-10 with POEC, and 51.4, 41.2, 31.1, and 15.5% in TCM-HEPES with POEC, respectively. The above development rates to morula/blastocyst stages were significantly higher than those of the embryos cultured in the Ham's F-10 and TCM-HEPES with out POEC(P<0.05). The in vitro development rates to the morula/blastocyst stage of 1-cell embryos cultured in Ham's F-10 and TCM-HEPES without POEC were 1.1∼1.2%. Especially, most of embryos were observed to arrest the development beyond 4-cell stages. As shown in the above results, the co-culture of in vitro produced porcine embryos with POEC in the two different media enhanced the development of fertilized eggs to morula/blastocyst stages in vitro. However, we didn't find out any difference for the in vitro development to morula/blastocyst stages between Ham's F-10 and TCM-HEPES media.

• Journal of Plant Biotechnology
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• v.29 no.1
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• pp.15-18
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• 2002

Callus and shoot formation of leaf explants from in vitro propagated shoots and field grown plants depending on the position of leaf, and four boxthron cultivars were investigated. Callus formation of explants from both in vitro shoot and field grown plants as easily achieved at the cut surfaces of explants but the callus formed from leaf of in vitro shoots was hardened as the duration of culture was proceed. Calli were effectively induced from leaves detached from the middle position of both in vitro and in vitro plants on MS medium containing 0.5 mg/L NAA with 0.2 mg/L BA, and the growth of calli were better in field grown leaves than in vitro grown leaves. Shoot formation were effectively induced from leaves detached from the upper position in vitro plants, and the middle parts of in vitro plants on MS medium containing 0.01 mg/L (NAA with 0.2 mg/ BA. There was difference on the frequency of shoot formation among four different cultivars; 'Jindojaerae' was the best for shoot formation followed by 'Cheonyang', 'Younghagukija' and 'Cheongyangjaerae'.

• Journal of Embryo Transfer
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• v.11 no.3
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• pp.241-248
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• 1996

This study was conducted to improve the production efficiency of in vitro produced (IVP) embryos in Korean Native cows. The optimal conditions and procedures for in vitro maturation(IVM), in vitro fertilization(IVF) and in vitro culture(IVC) of bovine follicular oocytes and IVP embryos were evaluated. Immature follicular oocytes were collected fiom the follicles of bovine ovaries obtained from abattoirs. The oocytes of Grade I and II for IVM were cocultured with monolayered bovine oviductal epithelial cells(BOEG) or granulosa cells in TCM-199 solution supplemented with follicle stimulating hormone, lutenizing hormone, estradiol-17$\beta$ and heat inactivated fetal calf serum at 39$^{\circ}C$ under 5% $CO_2$ in air for 14 to 24 hours. Most of the oocytes(93%) matured to metaphase II in 24 hours. The cocultured IVM oocytes were fertilized in vitro at significantly(P<0.05) higher rate with BOEC(83.8%) and with granulosa cells(84.6%) than the non-cocultured IVM oocytes(73.6%). The IVM-IVF embryos developed to morula and blastocyst at significantly(P<0.05) higher rate in coculture with BOEC(41.2%) than with granulosa cells(23.1%) or conditioned medium(23.4%).

• Journal of the Korean Home Economics Association
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• v.24 no.1
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• pp.43-51
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• 1986

In order to study antioxidative acton of garlic, alliin, scordinin, garlic oil, ethanol fraction and non-kaolin fraction which have been discovered from garlic until now were extracted and isolated, and each fraction was utilized as the experimental materials. Antioxidative action of each fraction was compared through the in vitro and in vivo experiments. Electron donatingability on $\alpha$, $\alpha$-diphenyl-$\beta$picrylhydra-$\chi$yl, the inhibitory effect of lipoperoxide formation by TBA and peroxide value were measured and analyzed. RESULTS : 1. When observed antioxidative ability by EDA value, ethanol fraction of garlic components showed the strongest reaction as 15.25. 2. In vitro experiment with TBA value, garlic oil, alliin and ethanol fracton showed distinctive effect on inhibitory effect of lipoperoxide formation. 3. Comparing with the inhibitory effect of lipoperoxide formation with TBA value in vivo, the ethanol fraction was the most effective in the blood or liver by intraperitoneal administration, whereas the ethanol fraction in the blood and non-kaolin fraction in the liver was most effective each other by orally administration. 4. In vitro experiment with peroxide value, garlic oil was distinctive effect on the inhivitory effect of lipoperoxide formation, which was a similar to the trend of TBA value in vitro. 5. Examining the induction time for the first period of lipoperoxide formation in vitro, garlix oil, ethanol fraction and alliin were effective, which was a similar to the trend of TBA value and peroxide value in vitro.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.47-47
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• 2003

The present study was conducted to determine the expression of the antioxidant enzyme(CuZn-SOD, Mn-SOD and GPX and apoptosis gene(caspase-3) for in vitro culture in in vitro maturation and in vitro fertilization(IVM/IVF) embryos in porcine. Porcine embryos derived from IVM/IVF were cultured in NCSU23 medium under 5% $CO_2$ in air at 38.5$^{\circ}C$. The patterns of gene expression for several antioxidant enzyme and apoptosis genes during preimplantion porcine embryo development were examined by the modified semi-quantitative single cell reverse transcriptase- polymerase chain reaction (RT-PCR). Preimplantation porcine embryos produced by IVM/IVF have expressed mRNAs for CuZn-SOD and GPX, whereas transcripts for Mn-SOD have not detected at any developmental stages. Expression of caspase-3 mRNA was detected at 2 cell, 8 cell, 16 cell and morula stages. The fas ligand transcripts were detected in porcine blastocyst. These results suggest that various antioxidant enzymes and apoptosis genes play crucial roles in in vitro culture of porcine IVM/IVF embryos.

• Asian-Australasian Journal of Animal Sciences
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• v.9 no.4
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• pp.371-374
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• 1996

Experiments were conducted to determine the effect of time intervals between collecting and use of cattle faeces as a source of micro-organisms in in vitro digestibility assays of forages. The results suggested that temperature conservation capacity by faeces depended on the size of the sample. There was no significant difference(p>0.05) between the first (T1 or 08:30 h) and second using time (T2 or 10:30 h). In vitro organic matter digestibility was significantly lower when faeces was used 5 h (T3 or 13:30 h) after collection. However, the organic matter digestibility determined at the second using time (T2) and third using time (T3) were highly correlated ($R^2=0.99$) with the first using time. It was concluded that faeces can be used as a source of microorganisms for in vitro digestibility assays of forages even 5 h after being voided.