• Title/Summary/Keyword: In Vitro Experiment

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Utility of Structural Information to Predict Drug Clearance from in Vitro Data

  • Lee, So-Young;Kim, Dong-Sup
    • Interdisciplinary Bio Central
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    • v.2 no.2
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    • pp.3.1-3.4
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    • 2010
  • In the present research, we assessed the utility of the structural information of drugs for predicting human in vivo intrinsic clearance from in vitro intrinsic clearance data obtained by human hepatic microsome experiment. To compare with the observed intrinsic clearance, human intrinsic clearance values for 51 drugs were estimated by the classical methods using in vivo-in vitro scale-up and by the new methods using the in vitro experimental data and selected molecular descriptors of drugs by the forward selection technique together. The results showed that taking consideration of molecular descriptors into prediction from in vitro experimental data could improve the prediction accuracy. The in vitro experiment is very useful when the data can estimate in vivo data accurately since it can reduce the cost of drug development. Improvement of prediction accuracy in the present approach can enhance the utility of in vitro data.

In vitro Fertilization and Culture of Rabbit Egg (가토난자의 체외수정과 체외배양)

  • 박흠대;이경광;정길생
    • Korean Journal of Animal Reproduction
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    • v.4 no.1
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    • pp.75-82
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    • 1980
  • This experiment was carried out to improve a simple and effective procedure of egg transfer which is considered to be the most useful technique for the improvementand proliferation of domestic animals. Several experiment procedures such as superovulation, surgical recovery of ovulated egg, in vitro capacitation of ejaculated spermatozoa, in vitro fertilization and culture of embryo were conducted. The results obtained in this experiment were summarized as follows: 1. In rabbits treated with PMS in combination with estradiol and HCG, 10 to 43 eggs were obtained in one rabbit and the average ovulation number of 20 rabbits was 21 eggs. 2. Six to 24 eggs (average 13 eggs) were recovered from the removed oviducts by the methods of flushing technique and the average recovery rate of 20 rabbits was 61.9 percent. 3. Most rabbit spermatozoa ejaculated by artificial vagina were capacitated in vitro by the culture of the spermatozoa with PBI medium at 37$^{\circ}C$ for 4 hours after removal of seminal plasma. 4. Normal fetilization following in vitro fetilization was observed in 88 (42.7%) of 206 eggs. 5. The number of eggs developed to 2-, 4-, and 8-cell stage following in vitro culture with PBI medium at 37$^{\circ}C$ was 26 (70.3%), 20 (54.1%) and 17 (45.9%) of 37 eggs used for in vitro culture.

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PREVENTION OF MURAL THROMBUS IN POROUS INNER TUBE OF DOUBLE-LAYERED TUBE BY SALINE PERFUSION (생리식염수의 재료표면에의 분출에 의한 이중튜브의 응혈 방지)

  • Kim, Sung-S.;Park, Joon-B.
    • Proceedings of the KOSOMBE Conference
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    • v.1993 no.11
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    • pp.110-113
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    • 1993
  • An in vitro experiment under laminar non-pulsatile blood flow and an acute canine ex vivo femoral A-V series shunt experiment were undertaken to investigate the effectiveness of saline perfusion through pores of porous tubes to prevent formation of mural thrombus. PS/SBR porous tubes were used for the in vitro experiment. Commercially obtained ePTFE porous tubes were etched by sodium naphthalenide, and the etched tubes were used for the ex vivo experiment. According to the results of the in vitro experiment, mural thrombus on the surface of the porous tribe could be prevented by the saline perfusion. Adhered blood cells decreased semi-logarithmically with increased perfusion rate (up to $0.022\;ml/min-cm^2$) of isotonic saline solution. According to results of the ex vivo experiment, mural thrombus decreased with increased perfusion rate (upto $0.060\;ml/min-cm^2$).

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EFFECTS OF OVARY TYPE, OOCYTE GRADE, HORMONE, SPERM CONCENTRATION AND FERTILIZATION MEDIUM ON IN VITRO MATURATION, FERTILIZATION AND DEVELOPMENT OF BOVINE FOLLICULAR OOCYTES

  • Im, K.S.;Kim, H.J.;Chung, K.M.;Kim, H.S.;Park, K.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.8 no.2
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    • pp.123-127
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    • 1995
  • In vitro embryo production (IVP) is affected by various factors during in vitro maturation, fertilization, and development. In this experiment, the effect of ovary type, quality of follicular oocyte, medium used for fertilization, presence of hormone in medium, sperm concentration on in vitro maturation and fertilization were examined for effective IVP. In vitro maturation was carried out using TCM-199 supplemented with 15% FCS and hormones in 5% $CO_2$ incubator for 24h. In vitro fertilization was performed with frozen-thawed sperm in modified mTALP medium containing 0.3% BSA, $10{\mu}g/ml$ heparin, and 5mM/ml caffeine for 24h. The fertilized embryos were co-cultured on monolayer of cumulus cells in TCM-199. When oocytes were collected from functionally active and inactive ovaries, maturation rate was 76.9 and 7.7%, respectively. When oocytes were classified morphologically to good and poor grades, maturation rate was 75 and 58.8%, respectively. FSH + LH + $E_2$ (86.4%) showed higher maturation rate than control (53.0%) and FSH (73%). The fertilization rate was 28.2, 100 and 91.7% in $1.6{\times}10^5$, $5.0{\times}10^5$ and $10.0{\times}10^5$ sperm concentration per ml. When oocytes were fertilized in mTALP and BO media, fertilization and cleavage rates of oocytes in mTALP were higher (84.3 and 56.9%) than those (67.4 and 23.3%) in BO medium. In this experiment, in vitro maturation, fertilization and development of oocytes were affected by type of ovary, grade of oocyte, hormones, sperm concentration and fertilization medium.

A Study on Culture Environments of In Vitro Matured/In Vitro Fertilized Bovine Embryos I. Influence of Somatic Cells, Growth Factors or Culture Media on In Vitro Maturation of Bovine Oocytes (소 체외수정란의 발생배양에 적합한 배양환경 조성 연구 I. 체세포, 성장인자 또는 배양액 종류가 난포란의 체외성숙에 미치는 효과)

  • 이명식;박수봉;박진기;장원경;민관식;백광수;성환후;박용윤
    • Korean Journal of Animal Reproduction
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    • v.22 no.1
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    • pp.95-99
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    • 1998
  • Three experiments were conducted with follicular oocytes, to compare some somatic cells, growth factors and media for in vitro maturation of bovine oocytes. In the first experiment, the type of somatic cells had no effects on in vitro maturation of bovine follicular ooctyes. In the second experiment, oocytes were matured in TCM199 su, pp.emented with growth factors on IVM of bovine follicular oocytes, then all were co-cultured with cumulus cells. The proportion of used oocytes that developed to expanding blastocysts was 22.2%, 20.2%, 17.7%, 22.2%, 24.4% and 20.2% after maturation in TCM199 su, pp.emented with control, insulin, IGF-I, IGF-Ⅱ, FGF and EGF, respectively. In the third experiment, oocytes were matured in BO, Ham's F10 and TCM199, then all were fertilized in BO, and embryos cultured in BO, Ham's F10 and TCM199, respectively. Cleavage rates in BO were 90%, had higher than in Ham's F10(80%) or in TCM199(64%). But production of expanding blastocysts in TCM199(21%) or Ham's F10(20.6%), had higher than in BO(4.6%).

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Evaluation of Manganese Ion on Controlling Harmful Microorganisms In vitro and In vivo for the Early-Weaned Pig

  • Kim, I.H.;Kang, D.H.;Kim, C.S.;Seok, H.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.1
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    • pp.8-12
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    • 1998
  • Two experiments were conducted to determine the effects of $MnSO_4$ on controlling harmful microorganisms in vitro and in vivo. The in vitro experiment was conducted to examine the effects of manganese sulfate $(MnSO_4)$ on the reduction of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) by growth stimulation of Pediococcus acidilactici (P. acidilactici; lactic acid bacteria). Manganese ion (0.003 %) was found to stimulate the growth of P. acidilactici in the In Vitro system. When E. coli and S. aureus were grown in a mixture with P. acidilactici, their numbers were reduced. This may be the result of a reduction of pH in the medium as a result of better growth of P. acidilactici due to stimulation by the Mn ion. The in vivo experiment was conducted to determine the effects of $MnSO_4$ in diets on controlling harmful microorganisms in fecal samples of pigs. There were no significant differences for the microbial numbers (i.e., total microorganisms, E. coli, lactic acid bacteria and S. aureus) in feces of pigs fed $MnSO_4$ compared to feces of pigs fed the control diet through 7 days. However, on day 7 of experiment, the pH of feces in pigs fed $MnSO_4$ (0.1%) decreased faster than pigs fed the control diet.

Effect of Puerariae Radix Methanol Extract on Benzo(a)pyrenc -in - duced Hepatotoxicity in Rats (갈근 메탄올 엑기스가 흰쥐에 있어서 Benzo(a)pyrene에 의해 유도된 간장해에 미치 는 영향)

  • 이윤경
    • Journal of the East Asian Society of Dietary Life
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    • v.4 no.2
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    • pp.59-67
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    • 1994
  • The present study was conducted to evaluate the hepatoprotective effect of puerariae Radix methanol extract on benzo(a) pyrene(B(a)P) - induced liver injuries in rats. In vitro experiment, primary cultured hepatocytes (5X105 cells/$m\ell$) were cultured for 20~24 hours after adding puerariae Radix mehtanol extract(32$\mu\textrm{g}$/$m\ell$) and B(a)P(50 uM). In vivo experiment, Puerariae Radix methanol extract(0.25 g/kg/day, per os) was administered for 7 days and B(a)P(0.1 mg/kg/day, intraperitoneally) was given after the last administration of extract. And then the hepatoprotective effect of Puerariae Radix methanol extract was investigated biochemically through in vitro and in vivo experiments. Namely, activities of enzymes (GOT, GPT and LDH) were measured and 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide(MTT) assay were carried out in vitro cell culture study and GOT, GPT, LDH and ALP activities and HDL-cholesterol, total cholesterol and triglyceride contents were performed in vivo study. In vitro experiment, as a result of enzyme activity measurement(GOT, GPT and LDH) and MTT assay, GOT,GPT and LDH activities changed by B(a)P were recovered to normal levels and hepatocytes impaired by B(a)P were recovered to normal. In vivo experiment, Puerariae Radix methanol extract significantly decreased the enzyme activities(GOT, GPT, ALP and LDH in serum and GPT and ALP in tissue) and lipid contents in comparison to B(a)P-treated group.

Effects of Fungicides on Inhibition of in Vitro Strawberry Pollen Germination (In Vitro에서 살균제의 딸기 화분발아 억제 효과)

  • Nam, Myeong Hyeon;Kim, Hyun Sook;Choi, Je Hyun;Lee, He Duck
    • Horticultural Science & Technology
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    • v.31 no.5
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    • pp.633-639
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    • 2013
  • Fungicide applications are required to prevent the strawberry from Botrytis fruit rot and powdery mildew that infect open strawberry flowers, however, their effects of fungicides on pollen germination of strawberry have been rarely documented, particularly those from recently developed active fungicidal ingredients. In this study we have evaluated the effects of 24 commercial fungicidal formulations and 6 organic materials on pollen germination in 3 strawberry cultivars using in vitro assays. Pollens from strawberry had higher germination rates on agar with sucrose of 18% and $25^{\circ}C$ than other tested conditions. Pollen germination rates of cvs. Seolhyang, Maehyang, and Kumhyang at 18% sucrose and $25^{\circ}C$ were 15.3, 18.4 and 30.7%, respectively. Pyraclostrobin, azoxystrobin, kresoxim-methyl, dichlofluanid, iminoctadine tris, and sulfur showed the strongest inhibitory efficacy with the germination rates of more than 93.8% compared to the no-fungicide control. Germination was not significantly affected by simeconazole and procymidone. This in vitro germination study may provide information useful for selecting fungicides in flowering stage to strawberry farmers.

In Vitro Culture and Cryopreservation of Bovine Embryos Derived from Matured and Fertilized In Vitro (소 체외수정란의 실용화를 위한 체외배양과 동결보존에 관한 연구)

  • 양부근;정희태;김정익
    • Journal of Embryo Transfer
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    • v.10 no.1
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    • pp.53-63
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    • 1995
  • The effects of different protein sources (serum vs bovine serum albumin), growth factors (EGF and PDGF) and co-culture with various type of somatic cel1s (BOEC, MEF and BRL) on the in vitro development of in vitro matured / in vitro fertilized bovine oocytes were examined, and the viability of frozen/thawed embryos derived from IVM /IVF was examined. Cell numbers of blastocysts were also counted. In Experiment 1, CR$_1$aa with serum was superior to CR$_1$aa with BSA in producing morulae plus blastocysts from IVM /IVF oocytes(24.4% vs 30.4%, p>0.05). In Experiment 2, more morulae plus blastocysts(42.3%) were produced in CR$_1$aa containing long /ml EGF than in the control CR$_1$aa(33.3%). In Experiment 3, 2- to 8-cell embryos derived from IVM /IVF oocytes were randomly allotted to one of 4 culture groups : a) CR$_1$aa ; b) CR$_1$aa + ing /ml PDGF ; CR$_1$aa + Sng /ml PDGF ; CR$_1$aa + lOng /ml PDGF ; culture resulted in 21.3, 51.2, 41.4 and 45.9%(p<0.05), respectively, developing into morulae and blastocysts. In Experiment 4, 0 and Sng /ml PDGF added to CR$_1$aa coculture with BRL or BOEC yielded 47.5, 42.5, 33.8 and 41.6% morulae and blastocysts, respectively. In Experiment 5, the proportion of embryos into morulae and blastocysts was highest in CR$_1$aa with MEF coculture group(50.9%) compared to any other group(CR$_1$aa, 22.3%; CR$_1$aa+BRL, 32.9%; CR$_1$aa+BOEC, 33.8%, p>0.05). In Experiment 6, survival rate of blastocysts produced by in vitro fertilization when cryoprotectant was removed in 0.7M glycerol+0.7M sucrose and 0.7M sucrose solution for 10 min. after thawing at 2$0^{\circ}C$ (Exp. H, 58.8%) was slightly higher than when cryoprotectant was removed 10%, 6.7% and 3.3% glycerol for 10 min. after thawing at 37$^{\circ}C$ (Exp. I, 54.3%). These study indicate that growth factors and somatic cell co-culture can increase the proportion of embryos that develop into morulae and blastocysts without an increase in the cell number and frozen /thawed method employed this experiment was not different.

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Effects of Strains of Saccharomyces cerevisiae and Incubation Conditions on the In vitro Degradability of Yeast and Roughage

  • Ando, S.;Nishiguchi, Y.;Hayasaka, K.;Yoshihara, Y.;Takahashi, J.;Iefuji, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.3
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    • pp.354-357
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    • 2005
  • The in vitro degradability of yeast and the effect of yeast on the in vitro degradability of forage may differ in terms of the specific yeast strains or their incubation conditions. Thus in experiment 1, two strains of sake yeast (strainK7 and strainK9) and one strain of bakers' yeast (KY5649) were incubated in an aerobic condition. In experiment 2, aerobically or anaero bically incubated K7 was used for investigating the in vitro degradability of yeast, the effect of yeast on the in vitro degradability of forage, and the degradability of yeast by pepsin and pronase treatment. The in vitrodegradability of bakers' yeast was significantly (p<0.05) higher than those of sake yeasts. The in vitro degradability of anaerobically incubated yeast was significantly (p<0.01) higher than that of aerobically incubated yeast. The degradability of bakers' yeast by pepsin treatment was significantly (p<0.01) higher than that of the sake yeasts. The degradability of bakers' yeast by pronase treatment was slightly higher than that of the two sake yeasts, while the degradability of anaerobically incubated yeast by both enzymes, respectively, was significantly (p<0.01) higher than that of aerobically incubated yeast. The degradability of forages was increased significantly (p<0.05) by the addition of yeasts. The degradability of roughage by sake yeast tended to be higher than that by the bakers' yeast. The degradability of roughage was significantly (p<0.05) higher by anaerobically incubated yeast than by aerobically incubated yeast. Given the above results, it seems that in vitro degradability of yeast and the magnitude of the increment of roughage degradation differ among the yeast strains and their incubation conditions.