• 약학회지
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• 제39권3호
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• pp.337-345
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• 1995

The effects of ginkgolides(natural mixture of ginkgolides, ginkgolide B, ginkgolide C) and flavonoids(quercetin, kaempferol, myricetin), extracted from Ginkgo biloba, on ADP and PAF-induced platelet aggregation in vitro and ex vivo were investigated. In these experiments, both of ginkgolides and ginkgoflavonoids did not affect the ADP(5 $\mu{M}$) induced platelet aggregation in vitro. The IC$_{50}$ value on PAF (0.3 $\mu{M}$) induced platelet aggregation were 2.52 $\mu{M}$ (ginkgolide B) and 6.35 $\mu{M}$ (natural mixture of ginkgolides) and 2.80 $\mu{M}$ (mixture of ginkgolide B and quercetin). Oral administration of ginkgolide B (1 and 3 mg/kg) and quercetin (3 and 9 mg/kg) to rabbits inhibited ex vivo PAF induced platelet aggregation in a dose-dependent manner. Ginkomin-F tablets administered to the diabetic patients showed inhibitory activities on the ADP and PAF induced platelet aggregation in a dose and time dependent manner.

• 농업생명과학연구
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• 제43권6호
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• pp.67-75
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• 2009

본 연구는 한약재 (결명자, 계피, 산초, 감초) 추출물의 반추위내 발효와 미생물 활성에 대한 효과를 구명하기 위하여 수행되었다. In vitro 건물소화율은 계피와 산초 추출물 첨가구에서는 0시간대, 감초 추출물 첨가구에서는 3시간대에 대조구에 비해 현저히 (P<0.05) 낮았다. 한약재 추출물 첨가에 따른 발효 시간대별 휘발성 지방산 조성의 변화는 3시간대에서만 처리간 유의성이 인정되었는데, acetate 비율은 대조구가 천연 추출물 첨가구보다 유의적으로 높았으나, butyrate, isobutyrate, isovalerate 및 valerate은 대조구에서 가장 낮았다 (P<0.05). 미생물 성장율은 발효 3시간대에서 결명자 첨가구를 제외한 한약재 첨가구에서 대조구에 비해 유의적 (P<0.05)으로 높았으나 다른 발효 시간대에서는 차이가 없었다. 이상의 결과로부터 한약재로 사용되고 있는 계피, 산초 및 감초 추출물을 in vitro 반추위 배양액에 첨가하였을 때, 발효초기에 반추위 미생물의 활성을 억제하는 경향은 있었으나 미생물 성장에 대한 억제 효과는 없는 것으로 나타났다.

• 한국가축번식학회지
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• 제19권2호
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• pp.153-157
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• 1995

These studies were conducted to investigate the microbe control effect of antibiotics treatment in all media used for in vitro fertilization(IVF) embryo production. The bovine oocytes were recovered from follicles(2~5mm) and were cultrued for 22hrs at 38.5$^{\circ}C$ with 5% CO2 incubator. The contamination and development rate in vitro fertilized oocyte was evaluated everyday. The results were summerized as follow ; 1. Control, antibiotic-antimycotic solution(AAS, Gibco) 1%＋gentamycin, and AAS 1%＋kanamycin(Sigma, USA) treatment was contaminated with 72hrs. However Baytril and Kanamycin(Korea) added was not contaminated. 2. The blastocyst formation rate in Baytril supplementated 1, 2 and 3${mu}ell$/ml was 3.73, 1.28 and 0.00%, respectively. 3. The blastocyst formation in kanamycin added concentration of 50, 75 and 100$\mu\textrm{g}$/ml was 13.0, 9.4 and 3.49%, respectively.

• Journal of Nutrition and Health
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• 제33권1호
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• pp.80-85
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• 2000

This study was undertaken to investigate the inhibitory effects of propolis on the in vitro proliferation of human colon(HT-29) and hepatoma(HepG2) cancer cell lines. The growth of the HT-29 and HepG2 cells was respectively inhibited by the administration of propolis in a concentration response-dependent manner. The distributions of HT-29 and HepG2 cells cultured in the medium containing propolis were shifted to the smaller sizes, and then HT-29 and HepG2 cells were shrunken under microscopic observations. The progression of cell cycle from G1 to S phase was significantly inhibited by propolis in the HT-29 and HepG2 cell lines, respectively. Those observations suggest that propolis has anticancer effect against some of cancer cell lines in vitro. (Korean J Nutrition 33(1) : 80-85, 2000)

• 한국가축번식학회지
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• 제14권1호
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• pp.36-42
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• 1990

The effect of preincubation on in vitro maturation and fertility were investigated using preovulatory oocytes with and without cumulus cells obtained from superovulated ouot-bred ICR mice. Oocytes were recovered from fully grown folicle at 10 hr after hCG administration. A part of oocytes recovered were treated with the solution of 0.1% hyaluronidase to remove cumulus cells. Both intact and treated oocytes were then incubated for 0 to 6hr in mT6 medium containing 0.3% BSA. After incubation for various times, a part of oocytes were subjected to the investigation of nuclear maturation and the remaining oocytes were used fro the induction of in vitro fertilization by adding them into medium containing capacitated mice epididymal spermatozoa. Above all, the percentage of preovulatory oocytes at the stage of metaphase II after preincubation for 0, 2, 4 and 6hr was 15.8, 36.4, 47.5 and 66.7%, respectively, suggesting the in vitro maturation of oocytes during their incubation. On the other hand, fertilizatin rate of oocytes preincubated for 0, 2, 4 and 6hr with and without cumulus cells were 41.0, 58.7, 68.7 and 75.6%, and 50.0, 45.1, 37.8 and 39.2%, respectively. No significant differences in fertilization rate between preovulatory oocytes preincubated for 6hr with cumulus cells and naturally ovulated were observed. These results suggest that cumulus cells take very important role in maturtion of oocytes in vitro. The precentage of preovulatory oocytes developed to 2-cell stage in vitro fertilization following preincubation for 0 to 6hr with and without cumulus cells ranged from 48.5 to 82.4% and 36.9 to 56.1%, respectively. Also, the rates of oocytes developed to blastocyst in vitro fertilization after preincubation for 0 to 6hr with and without cumulus cells were 28.1, 39.3, 42.5 and 44.0% and 12.5, 32.6, 24.4 and 15.5%, respectively. From these results, it could be said that fertility of preovulatory oocytes with cumulus cells could be improved to the level of that of naturally ovulated oocytes by adquate preincubation in vitro. Cumulus cells may, therefore, affect in vitro maturation, fertilization and following development of oocytes by influencing zona hardening.

• Asian-Australasian Journal of Animal Sciences
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• 제9권3호
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• pp.247-254
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• 1996

Three stages of growth of Italian ryegrass (pre-blooming, P-B; early-blooming, E-B; and late-blooming, L-B) were used to evaluate the effect of maturity on in vitro digestion of dry matter, fiber components and gas production. The rumen digestibility and gas production values were obtained by incubation of each sample in the rumen fluid of sheep for 12, 24, 36, 48 and 72 hr, respectively. The results showed that digestibility of dry matter (DM) significantly reduced (p < 0.05) as advancing maturity of the grass. Similarly, the digestibility of neutral detergent fiber (NDF) and acid detergent fiber (ADF) also significantly decreased (p < 0.05) with advancing maturity at all incubation times. However, the effect of maturity on digestibility of cellulose and hemicellulose was only detected when the samples were incubated more than 36 hr, where L-B was lower than P-B and E-B. Potential digestibility of nutrients, the maximum digestibility attainable in the rumen theoretically, was also higher at P-B than those of E-B and L-B. The amount of gas produced by microbial fermentation was closely related to the extent of DM digestion, and it was negatively correlated with advancing maturity of the grass.

• Reproductive and Developmental Biology
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• 제32권4호
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• pp.237-243
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• 2008

The objective of this study was to examine the effect of caffeine and sodium bicarbonate in a fertilization medium on the fertilizability of boar spermatozoa that were frozen in straws. Boar spermatozoa were extended with Beltsville F5 extender and frozen in 0.25-ml straws. In vitro matured porcine oocytes were fertilized in vitro (IVF) with frozen-thawed boar spermatozoa for 6h in a modified tris-buffered medium (mTBM) or in its modified medium by substituting the tris with 25mM sodium bicarbonate (modified bicarbonate-buffered medium; mBBM). Some of inseminated oocytes were fixed and stained for examination of sperm penetration. IVF embryos were cultured in a North Carolina State University-23 medium for embryo development. The percentage of live sperm was $47{\pm}4%$ and morphological abnormality of acrosome was found in $14{\pm}3%$ of spermatozoa. Optimal sperm concentration for IVF was $0.75{\sim}1.0{\times}1.0{\times}10^6$ sperms/ml when mTBM containing 5mM caffeine was used as the fertilization medium. Sperm penetration was significantly (p<0.05) stimulated by increasing caffeine concentration in the IVF medium. In addition, mBBM significantly (p<0.05) increased sperm penetration (92%) compared to mTBM (65%). More (p<0.05) blastocysts (22% vs. 32%) developed from the oocytes that were fertilized in mBBM containing 1mM caffeine than from those fertilized in mTBM with 5mM caffeine. Our results indicate that boar spermatozoa can be frozen successfully in straws with holding their normal fertilizability and that caffeine and sodium bicarbonate stimulates sperm penetration in vitro.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.43-43
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• 2001

This study evaluated the effect of fertilization-promoting peptide (FPP) on fertilizing ability and glycosidase activity in vitro of spermatozoa frozen-thawed in pig, Using chlortetracycline fluorescence analysis, the various glycosidase analyses and the oocyte penetration test, we have obtained evidence that FPP can promote the fertilizing ability and glycosidase activity of frozen-thawed spermatozoa in vitro. When frozen-thawed spermatozoa was washed with different concentrations of FPP, there were significantly (P<0.05) more acrosome-reacted in medium with 100 nM than 0, 50, 200 and 400 nM. The penetration rates were also highest in medium containing with 100 nM FPP (P<0.05). On the other hand, the $\beta$-N-acetylglucosaminidase activity was at least twofold higher than other glycosidase. In same glycosidase, however, there were no difference in medium with different concentrations of FPP In another experiment, spermatozoa preincubated in medium with or without FPP for 0, 1, 2, 3 and 4 h were inseminated with oocytes matured in vitro. The percentages of spermatozoa that reached acrosome reaction were affected by preincubation and were higher in medium with that than without FPP. When oocytes were inseminated with spermatozoa preincubated in medium with and without FPP during the different periods, however, penetration rates were decreased with preincubation periods of spermatozoa. On the other hand, when the sperm-oocyte were cultured for 4, 8, 12, 16, 20 and 24 h, the penetration rates were higher in spermatozoa preincubated with that than without FPP and had a tendency to increase as time of culture periods. However, The activities of $\alpha$-fucosidase, $\alpha$ -mannosidase, $\beta$-galactosidase and N-acetyl- $\beta$-D-glucosaminidase were higher in medium with that than without FPP regardless of periods of sperm preincubation and sperm-oocyte culture. These results suggest that FPP may play a positive role in promoting of sperm function and glycosidase activity in vitro in pig.

• 한국가축번식학회지
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• 제20권2호
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• pp.163-170
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• 1996

The effect of several potential antioxidants and amino acids were examined as a means of increasing the development of in vitro matured and in vitro fertilized oocytes into morulae or blastocysts. Bovine embryos developed to the 2~8 cell stage after in vitro fertilization were cultured for 5 to 6 days at 39$^{\circ}C$ in CR1aa containing varing concentraton of the antioxidants and amino acid in a gas phase consisting of 5% CO2, high humidified air. At 5~6 days, embryo developments were reduced, and embryos were fixed and stained with Hochest 33342 DNA stain to facilitate counting of cells. In experiment 1, the proportion of embryos developed to morulae and blastocysts in CR1aa containing 1mM, 2.5mM taurine (22.6% and 20.4%) was slightly higher than those of 0, 5 and 10mM Taurine (5.7, 5.7 and 3.9%, P<0.05). In experiment 2, addition of glutathione did not improve blastocyst development (P>0.05). In experiment 3, concentations of superoxide dismutase(SOD) ranging from 300 to 1,000 U did not affect the propotion of embryos developing into blastocysts (P>0.05). In experiment 4, addition of 250 U catalase(38.5%) was slighty higher than those of 0, 500 and 1,000U. In experiment 5, the proportion of embryo developed beyond morula stage in CR1aa with taurine plus EDTA was slighty higher than other treatments(15.7, 26.0 and 29.2%), there were no significantly increases in cell number among treatments(P>0.05). These results are indicating that antioxidants and amino acids can increase the proportion of embryos that develop into morulae and blastocysts, but did not increas in cell number of blastocysts.

• Journal of Applied Biological Chemistry
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• 제44권2호
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• pp.71-76
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• 2001

Methanol extract and solvent fractions of Aster scaber Thunb. were tested to elucidate antioxidative characteristics in vitro. Methanol extract showed strong inhibition on linoleic acid autoxidation and conjugated diene production. Hexane and diethyl ether fractions showed stronger chelating effect on $10^{-4}M\;FeCl_3$ than the other fractions. Butanol fraction had the strongest capacity on $10^{-3}M\;CuSO_4$ similar to ${\alpha}$-tocopherol. Synergistic effect of butanol fraction with histidine, methionine, and lysine on linoleic acid autoxidation had been observed. Contents of total phenol, vitamin E, vitamin C, and carotenoid were evaluated. From the results, Aster scaber Thunb. could be expected to play a role as an antioxidant in vivo.