<i>In vitro</i> inhibitory effects of Wen‐pi‐tang‐Hab‐Wu‐ling‐san on human cytochrome P450 isoforms

Lee, H. W.,Kim, D. W.,Phapale, P. B.,Lim, M. ‐,S.,Park, J.,Seo, J. J.,Park, K. M.,Park, Y. ‐,K.,Yoon, Y. ‐,R. Blackwell Publishing Ltd 2011 Journal of clinical pharmacy and therapeutics Vol.36 No.4

<P><B>Summary</B></P><P><B>What is known and Objective: </B> Although Wen‐pi‐tang‐Hab‐Wu‐ling‐san (WHW), an oriental herbal medicine, has been prescribed for the treatment of chronic renal failure (CRF) in Korean clinics, no studies regarding WHW–drug interactions had been reported. The purpose of this study was to evaluate the possibility that WHW inhibits the catalytic activities of major cytochrome P450 (CYP) isoforms.</P><P><B>Methods: </B> The abilities of various WHW extracts to inhibit phenacetin O‐de‐ethylation (CYP1A2), tolbutamide 4‐methylhydroxylation (CYP2C9), omeprazole 4′‐hydroxylation (CYP2C19), dextromethorphan O‐demethylation (CYP2D6), chlorzoxazone 6‐hydroxylation (CYP2E1) and midazolam 1‐hydroxylation (CYP3A4) were assessed using human liver microsomes.</P><P><B>Results and Discussion: </B> WHW extract at concentrations up to 100 μ<SMALL>m</SMALL> showed negligible inhibition of the six CYP isoforms tested (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and CYP3A4), with apparent IC<SUB>50</SUB> values (concentration of the inhibitor causing 50% inhibition of the original enzyme activity) of 817.5, 601.6, 521.7, 310.2, 342.8 and 487.0 μg/mL, respectively.</P><P><B>What is new and Conclusion: </B> Our <I>in vitro</I> findings suggest that WHW extract at concentrations corresponding to a clinically recommended dosage range has no notable inhibitory effects on CYP isoforms. Therefore, we believe that WHW extract may be free of drug–herb interactions when co‐administered with other medicines. However, <I>in vivo</I> human studies are needed to confirm these results.</P>

Efficient long-term amplification of hepatitis B virus isolates after infection of slow proliferating HepG2-NTCP cells

Kö,nig, Alexander,Yang, Jaewon,Jo, Eunji,Park, Kyu Ho Paul,Kim, Hyun,Than, Thoa Thi,Song, Xiyong,Qi, Xiaoxuan,Dai, Xinghong,Park, Soonju,Shum, David,Ryu, Wang-Shick,Kim, Jung-Hee,Yoon, Seung Kew,P Elsevier 2019 Journal of hepatology Vol.71 No.2

<P><B>Background & Aims</B></P> <P>As hepatitis B virus (HBV) spreads through the infected liver it is simultaneously secreted into the blood. HBV-susceptible <I>in vitro</I> infection models do not efficiently amplify viral progeny or support cell-to-cell spread. We sought to establish a cell culture system for the amplification of infectious HBV from clinical specimens.</P> <P><B>Methods</B></P> <P>An HBV-susceptible sodium-taurocholate cotransporting polypeptide-overexpressing HepG2 cell clone (HepG2-NTCPsec+) producing high titers of infectious progeny was selected. Secreted HBV progeny were characterized by native gel electrophoresis and electron microscopy. Comparative RNA-seq transcriptomics was performed to quantify the expression of host proviral and restriction factors. Viral spread routes were evaluated using HBV entry- or replication inhibitors, visualization of viral cell-to-cell spread in reporter cells, and nearest neighbor infection determination. Amplification kinetics of HBV genotypes B-D were analyzed.</P> <P><B>Results</B></P> <P>Infected HepG2-NTCPsec+ secreted high levels of large HBV surface protein-enveloped infectious HBV progeny with typical appearance under electron microscopy. RNA-seq transcriptomics revealed that HBV does not induce significant gene expression changes in HepG2-NTCPsec+, however, transcription factors favoring HBV amplification were more strongly expressed than in less permissive HepG2-NTCPsec−. Upon inoculation with HBV-containing patient sera, rates of infected cells increased from 10% initially to 70% by viral spread to adjacent cells, and viral progeny and antigens were efficiently secreted. HepG2-NTCPsec+ supported up to 1,300-fold net amplification of HBV genomes depending on the source of virus. Viral spread and amplification were abolished by entry and replication inhibitors; viral rebound was observed after inhibitor discontinuation.</P> <P><B>Conclusions</B></P> <P>The novel HepG2-NTCPsec+ cells efficiently support the complete HBV life cycle, long-term viral spread and amplification of HBV derived from patients or cell culture, resembling relevant features of HBV-infected patients.</P> <P><B>Lay summary</B></P> <P>Currently available laboratory systems are unable to reproduce the dynamics of hepatitis B virus (HBV) spread through the infected liver and release into the blood. We developed a slowly dividing liver-derived cell line which multiplies infectious viral particles upon inoculation with patient- or cell culture-derived HBV. This new infection model can improve therapy by measuring, in advance, the sensitivity of a patient’s HBV strain to specific antiviral drugs.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Cell culture system that mimicks complete HBV life cycle from entry to egress. </LI> <LI> Efficient <I>in vitro</I> infection with crude HBV patient sera. </LI> <LI> Up to 50- and 1,300-fold net amplification of patient- and cell culture-derived input HBV in the supernatant. </LI> <LI> Polyethylene glycol-independent HBV spread to adjacent cells, forming infected cell clusters. </LI> <LI> Evaluation of patient- and cell culture-derived HBV amplification w/wo antivirals over 8 weeks. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Effect of <i>CYP3A5*3</i> genotype on serum carbamazepine concentrations at steady-state in Korean epileptic patients

Park, P.-W.,Seo, Y. H.,Ahn, J. Y.,Kim, K.-A.,Park, J.-Y. Blackwell Publishing Ltd 2009 Journal of clinical pharmacy and therapeutics Vol.34 No.5

<P>Abstract</P><P>Background and Objective: </P><P>Carbamazepine (CBZ) is metabolized mainly by the CYP3A family of enzymes, which includes CYP3A4 and CYP3A5. Several studies have suggested that the <I>CYP3A5*3</I> genotype influences the pharmacokinetics of CYP3A substrates. The present study aimed to assess the effect of the <I>CYP3A5*3</I> genotype on serum concentration of CBZ at the steady-state in Korean epileptic patients.</P><P>Method: </P><P>The serum concentrations of CBZ in 35 Korean epileptic patients were measured and their <I>CYP3A5</I> genotype was determined. Fourteen patients were <I>CYP3A5</I> expressors (two for <I>CYP3A5*1/*1</I> and 12 for <I>CYP3A5*1/*3</I>) and 21 patients were <I>CYP3A5</I> non-expressors (<I>CYP3A5*3/*3</I>). Dose-normalized concentrations (mean ± SD) of CBZ were 9·9 ± 3·4 ng/mL/mg for <I>CYP3A5</I> expressors and 13·1 ± 4·5 ng/mL/mg for <I>CYP3A5</I> non-expressors (<I>P</I> = 0·032). The oral clearance of CBZ was significantly higher in <I>CYP3A5</I> non-expressors than that of <I>CYP3A5</I> expressors (0·056 ±0·017 L/h/kg vs. 0·040 ± 0·014 L/h/kg, <I>P</I> = 0·004). The <I>CYP3A5</I> genotype affected the CBZ concentrations in Korean epileptic patients and is a factor that may contribute to inter-individual variability in CBZ disposition in epileptic patients.</P>

Comparison of 90‐day case‐fatality after ischemic stroke between two different stroke outcome registries using propensity score matching analysis

Yu, K‐,H.,Hong, K‐,S.,Lee, B‐,C.,Oh, M‐,S.,Cho, Y‐,J.,Koo, J‐,S.,Park, J‐,M.,Bae, H‐,J.,Han, M‐,K.,Ju, Y‐,S.,Kang, D‐,W.,Appelros, P. Blackwell Publishing Ltd 2011 Acta neurologica Scandinavica Vol.123 No.5

<P>Yu K‐H, Hong K‐S, Lee B‐C, Oh M‐S, Cho Y‐J, Koo J‐S, Park J‐M, Bae H‐J, Han M‐K, Ju Y‐S, Kang D‐W, Appelros P, Norrving B, Terent A. Comparison of 90‐day case‐fatality after ischemic stroke between two different stroke outcome registries using propensity score matching analysis. 
Acta Neurol Scand: 2011: 123: 325–331. 
© 2010 John Wiley & Sons A/S.</P><P><B>Background – </B> It has not been clarified whether the disparity in ischemic stroke outcome between populations is caused by ethnic and geographic differences or by variations in case mix. Propensity score matching (PSM) analysis can overcome some analytical problems but is rarely used in stroke outcome research. This study was to compare the ischemic stroke case‐fatality between two PSM cohorts of Sweden and Korea.</P><P><B>Methods – </B> Prognostic variables related to baseline characteristics and stroke care were included in our PSM model. Then, we selected 7675 Swedish and 1220 Korean patients with ischemic stroke from each stroke registers and performed one‐to‐one matching based on propensity scores of each patient.</P><P><B>Results – </B> After PSM, all measured variables were well balanced in 1163 matched subjects, and the 90‐day case‐fatality was identical 6.2% (HR 0.997, 95%CI 0.905–1.099) in Sweden and Korea.</P><P><B>Conclusions – </B> No difference is found in the 90‐day case‐fatality in propensity score‐matched Swedish and Korean patients with ischemic stroke.</P>

Reproductive Performance, Milk Composition, Blood Metabolites and Hormone Profiles of Lactating Sows Fed Diets with Different Cereal and Fat Sources

Park, M.S.,Shinde, P.L.,Yang, Y.X.,Kim, J.S.,Choi, J.Y.,Yun, K.,Kim, Y.W.,Lohakare, J.D.,Yang, B.K.,Lee, J.K.,Chae, Byung-Jo Asian Australasian Association of Animal Productio 2010 Animal Bioscience Vol.23 No.2

Different dietary cereal sources and fat types in the lactation diet were evaluated to investigate their effects on reproductive performance, milk composition, blood metabolites and hormones in multiparous sows. Twenty-four sows were randomly assigned to one of four treatments according to a 2${\times}$2 factorial arrangement of treatments. Each treatment had 6 replicates comprising 1 sow. Two cereal (corn or wheat) and two fat (tallow or soybean oil) sources were used to prepare iso-caloric and iso-nitrogenous diets. Sows fed corn-based diets lost less body weight (p = 0.003) and backfat thickness (p = 0.034), consumed more feed (p = 0.032) and had shorter wean-to-estrus interval (p = 0.016) than sows fed wheat-based diets. Fewer piglets and lower body weight of piglets (p<0.05) at weaning were noted in sows fed wheat-based diets than in sows fed corn-based diets. However, no significant effects (p>0.05) of dietary fat source and its interaction with dietary cereal source on sow body condition and reproductive performance were observed during lactation. Feeding of a corn-based diet improved (p<0.05) sow milk total solid, protein and fat, increased blood urea nitrogen (p = 0.032) and triglyceride (p = 0.018), and decreased blood creatinine (p = 0.011) concentration at weaning when compared with sows fed wheatbased diets. Sows fed corn-based diets had higher concentration of insulin (p = 0.048) and LH (p<0.05) at weaning than sows fed wheatbased diets. The results indicate that feeding corn-based diets to lactating sows improved sow body condition and reproductive performance compared with wheat-based diets regardless of fat sources.

Effects of Deletion of Ca Supplement (limestone) on Growth and Beef Quality in Hanwoo Finishing Steers

Lee, C. E.,Park, N. K.,Seong, P. N.,Jin, S. H.,Park, B. Y.,Kim, K. I. 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.3

이 연구는 소 비육후기에 칼슘제(석회석)를 첨가하지 않은 사료의 급여가 성장율, 근내지방도 및 혈청 1,25-dihydroxy vitamin D_3(1,25(OH)_2D_3) 함량에 미치는 영향을 구명하기 위하여 수행되었다. 거세 한우 24두(20~24개월령)를 12두씩 대조구(석회석 2.5% 함유 농후사료)와 칼슘제 무첨가구(석회석 0%)로 배치하여 223일 동안 사료(농후사료 및 오차드그라스 건초)와 물을 무제한 급여하였고, 사양시험이 완료된 후 도축하여 육질을 평가하였다. 혈청 Ca^2+, Ca 및 P 함량에는 처리 간 차이가 없었으나 (P>0.05), 1,25(OH)_2D_3 함량은 시험 시작후 2 또는 6개월째 모두 칼슘제 무첨가구가 대조구보다 (각각 78.3 vs 51.7 또는 80.3 vs 51.1pg/mL) 높았다 (P<0.01). 칼슘제를 첨가하지 않은 사료를 급여한 비육우가 대조구보다 농후사료 섭취량은 증가하고 건초 섭취량은 감소하는 경향을 보였다. 일당증체량은 대조구보다 칼슘제 무첨가구에서 높았다(P<0.01). 등심단면적(82.8 vs 77.2㎠), 근내지방도(5.1 vs 2.2) 및 지방 함량(10.2 vs 6.7%)이 칼슘제 무첨가구가 대조구보다 높았고 (P<0.05), 수분 함량 (67.6 vs 70.4%)은 낮았다 (P<0.05). 등심 육색, pH 및 보수력에서는 처리 간 차이가 없었으나 전단력에서는 칼슘제 무첨가구에서 (2.9 vs 3.2 kg/1.27-㎝ diameter core), 약간 낮게 (P = 0.08) 나타났다. 관능평가에서는 칼슘제 무첨가구가 대조구보다 연도 (4.9 vs 4.5) 및 향미 (4.9 vs 4.6)가 약간 개선되었으나 (P<0.05) 다즙성에서는 처리간 차이가 없었다 (P<0.05) 다즙성에서는 처리간 차이가 없었다 (P<0.05). 본 연구결과는 비육후기에 칼슘제(석회석)를 첨가하지 않은 사료의 급여는 에너지 섭취의 증가 또는 1,25(OH)_2D_3의 합성 촉진을 통하여, 근내지방합성이 증가되어 성장율 및 근내지방도를 개선한다는 것을 제시하였다. A study was conducted to determine the effects of feeding a diet containing no Ca supplement (limestone) during the late finishing period on growth, marbling and serum 1,25-dihydroxy vitamin D_3 level in Korean native cattle. Twenty-four steers (20~24 mo of age) were divided into two groups of 12 each: one group assigned to a control diet (concentrates containing 2.5% limestone) and the other to a diet containing no calcium supplement. They were allowed to have free access to diets (concentrates and orchard grass hay) and water during the entire feeding period (223 d). Serum Ca^2-, Ca and P concentrations were not influenced by diets, but serum 1,25-dihydroxy vitamin D_3 concentrations determined 2 or 6 mo after the beginning of feeding the experimental diets were higher (P<0.01) in steers fed the diet without Ca supplement than in those fed the control diet (78.3 vs 51.7 and 80.3 vs 51.1 pg/mL, respectively). Steers fed the diet without Ca supplement tended to have a higher intake of concentrates, but a lower intake of hay, compared to those fed the control diet. Average daily gain was higher (P<0.05) in steers fed the diet without Ca supplement than in those fed the control diet. Feeding the diet without Ca supplement remarkably (P<0.01) increased the marbling score (5.1 vs 2.2) and the muscle (M. longissimus dorsi) fat content (10.2 vs 6.7%) with a concomitant decrease in moisture content (67.6 vs 70.4%), compared to feeding the compared to feeding the control diet. Ribeye area was increased (77.2 vs 82.8 ㎤) with the diet without Ca supplement, compared to the control diet (P<0.05). Meat color, pH and water-holding capacity in longissimus muscle were not different between the two groups. The Warner-Brazler Shear (WBS) force of the longissimus muscle was slightly (P=0.08) lower in steers fed the diet without Ca supplement than in steers fed the control diet (2.9 vs 3.2 ㎏/1.27-㎝ diameter core). Sensory evaluation showed that feeding the diet without Ca supplement slightly (P<0.05) improved tenderness (4.9 vs 4.5) and flavor (4.9 vs 4.6), compared to feeding the control diet, but juiciness was not affected by diets. Results showed that deletion of Ca supplement fro finishing diets is beneficial, increasing growth and marbling partly through an increased energy intake and induced 1,25-dihydroxy vitamin D_3 synthesis that may increase intracellular Ca^2+ concentration and in turn fat synthesis.

Toward high efficiency organic photovoltaic devices with enhanced thermal stability utilizing P3HT-b-P3PHT block copolymer additives

Zhu, M.,Kim, H.,Jang, Y.,Park, S.,Ryu, D.,Kim, K.,Tang, P.,Qiu, F.,Kim, D.,Peng, J. Royal Society of Chemistry 2016 Journal of Materials Chemistry A Vol.4 No.47

<P>Organic photovoltaics (OPVs) have drawn an extensive amount of attention due to their low cost, processibility and flexibility. However, a cell based on a blend of poly(3-hexylthiophene) (P3HT) and [6,6]-phenyl-C-61-butyric acid methyl ester (PC61BM) has a limited power conversion efficiency (PCE) due to the short exciton diffusion length of similar to 10 nm. We address this issue by designing a series of all-conjugated diblock copolymers, poly(3-hexylthiophene)-b-poly(3-(6-diethylphosphonatohexyl) thiophene) (P3HT-b-P3PHT), intended for use as additives to improve the performance of P3HT:PC61BM-based photovoltaic devices. The PCE of the devices improved from 3.30% to 4.03% with the addition of P3HT-b-P3PHT (3 : 1). The thermal stability of devices with P3HT-b-P3PHT additives improved significantly relative to that of the P3HT:PC61BM reference device, where the devices including a copolymer with a higher P3PHT content exhibited a better thermal stability. It was found that the fill factor (FF) could be regulated by simply varying the block ratio of P3HT-b-P3PHT and played a crucial role in improving both the PCE and the thermal stability. The P3HT-b-P3PHT diffused at the P3HT:PC61BM interface, improved the miscibility between P3HT and PC61BM, optimized the nanoscale morphology of the photoactive layer, and reduced the active layer roughness, all of which improved the FF and thus contributed to an improvement in device performance.</P>

Variants in hormone-related genes and the risk of biliary tract cancers and stones: a population-based study in China

Park, S. K.,Andreotti, G.,Sakoda, L. C.,Gao, Y.-T.,Rashid, A.,Chen, J.,Chen, B. E.,Rosenberg, P. S.,Shen, M.-C.,Wang, B.-S.,Han, T.-Q.,Zhang, B.-H.,Yeager, M.,Chanock, S.,Hsing, A. W. Oxford University Press 2009 Carcinogenesis Vol.30 No.4

<P>Biliary tract cancers, encompassing gallbladder, extrahepatic bile duct and ampulla of Vater cancers, are uncommon but often fatal malignancies. Hormone-related factors, including parity, oral contraceptive use, obesity, and gallstones, have been implicated in the etiology of these cancers. To further clarify the role of hormones in biliary tract cancers and biliary stones, we genotyped 18 single-nucleotide polymorphisms (SNPs) in nine genes involved in steroid hormone biosynthesis, metabolism and transport in a population-based case-control study in Shanghai, China. This study included subjects who completed an interview and provided blood, which totaled 411 biliary tract cancer and 893 biliary stone patients and 786 healthy Shanghai residents. The CYP1A1 IVS1 + 606 (rs2606345) T allele was associated with gallbladder [odds ratio (OR) = 2.0, 95% confidence interval (CI), 1.3-3.0] and bile duct cancers (OR = 1.8, 95% CI = 1.1-3.1), whereas the CYP1A1 Ex7 + 131 (rs1048943) G allele was associated with ampulla of Vater cancer (OR = 2.9, 95% CI = 1.5-5.4). After taking into account multiple comparisons for SNPs within each gene, CYP1A1 was significantly associated with gallbladder (P = 0.004) and ampulla of Vater cancers (P = 0.01), but borderline with bile duct cancer (P = 0.06). The effect of CYP1A1 IVS1 + 606 on gallbladder cancer was more pronounced among non-obese (body mass index < 23) (OR = 3.3, 95% CI = 1.8-6.1; P interaction = 0.001). Among women taking oral contraceptives, the effect of SHBG Ex8 + 6 (rs6259) on gallbladder cancer (OR = 6.7, 95% CI = 2.2-20.5; P interaction = 0.001) and stones (OR = 2.3, 95% CI = 1.1-4.9; P-interaction = 0.05) was statistically significant. Our findings suggest that common variants in hormone-related genes contribute to the risk of biliary tract cancers and stones, possibly by modulating hormone metabolism.</P>

The valley Hall effect in MoS₂ transistors

Mak, K. F.,McGill, K. L.,Park, J.,McEuen, P. L. American Association for the Advancement of Scienc 2014 Science Vol.344 No.6191

<P><B>Using the valleys in monolayer MoS<SUB>2</SUB></B></P><P>The electronic structure of the two-dimensional material MoS<SUB>2</SUB> has two distinct “valleys” of energy that may help to carry information in future electronic devices. Mak <I>et al.</I> observed the so-called valley Hall effect in a monolayer of MoS<SUB>2</SUB>. The electrons from different valleys moved in opposite directions across the sample, with one valley being overrepresented with respect to the other. The scientists achieved this by shining circularly polarized light on the material, which created an imbalance in the population of the two valleys. The findings may enable practical applications in the newly formed field of valleytronics.</P><P><I>Science</I>, this issue p. 1489</P>

抗卵丘細胞 抗體가 牛卵胞卵의 體外成熟과 受精 및 豚發達에 미치는 影響

鄭吉生,朴世必,鄭炯敏,嚴基鵬,李勳澤,鄭炳鉉 건국대학교 동물자원연구센터 1991 動物資源硏究誌 Vol.16 No.-

本 硏究에서는 卵巢內에 존재하는 다수의 未成熟 卵胞卵을 회수한 다음 이를 in vitro system에서 成熟, 受精 및 胚發達에 있어서 卵丘細胞(cumulus cell)가 미치는 영향과 그 기전을 조사하였다. 卵丘細胞에 대하여 좀더 구체적인 검토를 수행할 목적으로 牛卵胞卵에 대한 抗 卵丘細胞 抗體(Rabbit anti-bovine cumulus cell antibody)를 제조하여 Ammonium sulfate precipitation과 Protein A-sepharose CL-4B affinity chromatography의 의해 IgG만을 분리 정제한 다음, indirect ELISA와 Immunoprecipitation法에 의해 확인된 이들 각각의 抗體(intact cumulus cell antibody, solubilized cumulus cell antibody)를 牛卵胞卵의 體外成熟과 受精時 첨가하는 卵丘細胞가 體外成熟과 受精 및 갑發達에 미치는 영향을 조사하여 다음과 같은 결과를 얻었다. 1) 無處理 卵丘細胞와 溶解 卵丘細胞를 抗原으로 처리한 개체로부터 血淸을 채취하여 間接酸素免疫分析法으로 抗體 형성여부를 확인한 결과 1:12,800의 稀釋倍率에서도 두 抗血淸 處理區가 對照 血淸區에 비해 높은 O.D.價를 나타냄으로써 두 가지 형태의 抗原이 다같이 抗卵丘細胞 抗體를 생산하는 것으로 확인되었다. 2) Iodogn과 Immunoprecipitation를 이용하여 항체의 생성을 재차 確認하였는데 Iodogen method를 이용한 牛卵丘細胞의 ?? Iabelling은 25,000/10㎕ cmp이었다. 또한 ??-bovine cumulus cell과 각각의 抗血淸 反應 결과 생성된 Immunoprecipitates의 radioactivity와 O.D價를 조사하였던바, 無處理 혹은 溶解化된 卵丘細胞에서 공히 높은 抗體의 생성을 확인했으며 두 處理群을 비교해 볼 때 溶解化된 卵丘細胞 抗體가 無處理 卵丘細胞 抗體보다 더 높은 反應性과 O.D價를 가지고 있음을 알 수 있었다. 3) 無處理 卵丘細胞에 대한 抗血淸과 溶解化된 卵丘細胞에 대한 抗血淸으로부터 抗體를 분리해 냈을 때 정제된 Ig G의 O.D.價를 각각 1.19mg/ml 이상이었다. 4) 卵丘細胞가 부착된 卵胞卵과 제거된 卵胞卵에 無處理 卵丘細胞에 의해 생성된 抗體를 처리한 결과, Metaphase Ⅱ 段階에까지 발달한 卵胞卵은 卵丘細胞가 제거된 卵胞卵의 對照區를 포함한 모든 抗體 處理區의 경우 47.6∼59.1%로써 卵丘細胞가 부착된 卵胞卵의 對照區 78.8%에 비해 유의하게 낮았다. (P<0.01). 5) 卵丘細胞가 부착된 卵胞卵과 제거된 卵胞卵의 溶解 卵丘細胞에 의해 생성된 抗體를 처리한 결과, M Ⅱ 단계에까지 발달한 卵胞卵은 卵丘細胞가 제거된 卵胞卵의 對照區를 포함한 모든 抗體 處理區의 경우 47.6∼59.1%로써 卵丘細胞가 附着된 卵胞卵의 對照區 82.1%에 비해 有意하게 낮았다.(P<0.01). 抗卵丘細胞 抗體의 작용으로 인하여 이러한 卵丘細胞의 효과는 억제되는 결과로 미루어 볼 때 卵丘細胞는 體外成熟시 卵胞卵의 核成熟을 증진시키는 것으로 사료된다. 6) 卵丘細胞가 부착되었거나 또는 제거된 成熟 牛卵胞卵에 武處理 卵丘細胞에 대한 抗體를 처리한 다음 體外受精을 실시한 결과 卵丘細胞가 제거된 卵胞卵에 있어서 對照區를 포함한 모든 抗體 處理區의 受精率 45.0∼53.7%는 卵丘細胞가 부착된 卵胞卵에 있어서 對照區의 受精率 64.3%에 비해 다소 낮았다. 한편 雌性 및 雄性 前核이 동시에 형성된 牛卵胞卵의 비율은 卵丘細胞를 제거한 卵胞卵의 對照毆를 포함한 모든 抗體 處理區가 18.2∼38.9%로써 卵丘細胞가 부착된 卵胞卵에 있어서의 對照區 51.9%에 비해 有意하게 낮았다.(P<0.05). 7) 卵丘細胞가 부착되었거나 또는 제거된 成熟 卵胞卵에 溶解 卵丘細胞에 대한 抗體를 처리한 다음 體外受精을 실시한 결과 卵丘細胞가 제거된 卵胞卵에 있어서 對照區를 포함한 모든 抗體 處理區의 受精率 45.0∼52.5%는 卵丘細胞가 부착된 卵胞卵에 있어서 對照區의 受精率 62.8%에 비해 다소 낮았다. 또한 雌性 및 雄性 前核이 동시에 형성된 卵子의 비율은 卵丘細胞가 제거된 卵胞卵에 있어서의 對照區를 포함한 모든 부착된 卵胞卵가 26.3∼38.3%로써 卵丘細胞가 부착된 卵胞卵에 있어서의 對照區 55.6%에 비해 有意하게 낮았다.(P<0.05). 8) 抗體處理를 받은 牛卵胞卵의 체외발생을 조사하였던 바, 卵丘細胞가 제거된 卵胞卵에 있어서 對照區를 포함한 모든 抗體 處理區의 受精率이 8-細胞期, 16-細胞期 및 桑實胚期 혹은 胚盤胞期까지 발달한 비율은 7.1∼14.5, 2.9∼5.9 및 1.5%∼2.9%로써 卵丘細胞가 부착된 난자에 있어서 對照區의 18.6, 10.0 및 8.6%에 비해 낮은 胚發生率을 나타냈다. 抗卵丘細胞 抗體를 처리함으로써 卵丘細胞 작용이 억제된다는 本 硏究 결과는 卵丘細胞가 卵胞卵의 體外受精率과 前核形成率 및 胚發生能에 유의한 효과가 있다는 것을 시사한다. These experiments were carried out to investigate the effect of cumulus cells on in vitro maturation, fertilization, and subsequent development of follicular oocytes recovered from the bovine ovaries as well as to study the mechanisms involved in these processes. To examine the specific role of cumulus cell, antisera to bovine cumulus cell were produced Japanese Giant rabbit by repeated immunization of intact or solubilized bovine cumulus cell and purified IgG by the Ammonium sulfate precipitation and Protein A-Sepharose affinity chromatography. Then, these IgG of the bovine cumulus cell-specific antibodies were confirmed by indirect ELISA and Immunoprecipitation, and the effects of theses antibodies were investigated in vitro maturation, fertilizaiton, and development of bovine follicular oocytes. The results obtained in these experiment were summarized as follows: 1. The titer of the antibodies to cumulus cell determined by indirect ELISA using intact or solubilized bovine cumulus cell coasted plated was very high in both intact and solubilized cumulus cell. Especially, the optical density(O.D.) at 1: 12,800 dilution of antibodies was significantly higher than those of non-immunized control sera. 2. To reexamine the immunity of antibody, cumulus cells were labelled with I using the Iodogen method and the radioactivity of I into cumulus cell was shown approximately 25,000cpm/10㎕. The high radioactivity and O.D. value in immunoprecipitating reaction of I-bovine cumulus cell with anti-cumulus cell antisera were reconfirmed the existence of immuno-reactivity of antibody to cumulus cell. In addition, reactivities and O.D. values of solubilized cumulus cell antibodies were significantly higher than those of intact cumulus cell antibody. 3. O.D. values of the purified intact and solubilized cumulus cell antibodies were more than 1.19mg/ml. 4. When the follicular oocytes were treated with antibody to intact cumulus cell, the maturation rates of cumulus-enclosed and removed oocytes were ranged from 47.6 to 59.1%. These values are significantly lower than those(78.8%) fo follicular oocytes cultured without the antibody (p<0.01). 5. The maturation rates of cumulus-enclosed and removed oocytes treated with antibody against solubilized cumulus cell were ranged from 46.7 to 59.1%, showing significantly lower than those(82.1%) of oocytes cultured in antibody free medium(P<0.01). These results that the beneficial effect of cumulus cell to the oocyte maturation was inhibited by the action of antibody to cumulus cell suggest that cumulus cell enhances nuclear maturation of follicular oocytes. 6. When the follicular oocytes matured in vitro were treated with antibody to intact cumulus cell, the fertilization rates of cumulus intact and removed oocytes were ranged from 45.0 to 53.7%. These values are slightly lower than those(64.3%) of follicular oocytes not treated with the antibody, and increased frequencies of both male and female pronucler formations were detected in cumulus intact oocytes cultured in medium without antibody (P<0.05). 7. The fertilizaiton rates of cumulus intact and removed oocytes treated with antibody to solubilized cumulus cell were ranged from 45.0 to 52.5%, significantly lower than those(62.8%) of oocytes cultured in antibody free medium, and increased frequencies of ova with male and female pronuclei formations were found in the presence of cumulus cells(P<0.05). 8. The development rates of cumulus intact and removed oocytes to 8-, 16-cell, and morula or blastocyst stage after treatment of intact and solubilized cumulus cell antibody were ranged from 7.1 to 14.5 from 2.9, and from 1.5 to 2.9T, respectively. These rates were slightly lower than 18.6, 10.0 and 8.6% of cumulus intact oocytes cultured in medium without the antibody. The results that beneficial effects of cumulus cell to the pronuclear formation and embryo development were blocked by the action of antibody to cumulus cell suggest that cumulus cell significantly promotes normal fertilizaiton with proper pronuclear formation and embryo development.