돼지 卵子의 透明帶에 대한 單一클론抗體生産과 그 特性에 關한 硏究

金鐘培,劉永春,金昌圭,權五中,鄭盛元,鄭吉生 건국대학교 동물자원연구센터 1991 動物資源硏究誌 Vol.16 No.-

本 試驗은 單一클론抗體의 강한 特異性과 抗體性質의 不變性을 이용하여 發生學的 側面에서 哺乳動物 卵子의 透明帶의 機能과 構造를 이해하고, 또한 種特異的인 精子 受容體의 存在 및 生化學的 構造를 규명하기 위한 기초연구로서, 돼지 卵子의 透明帶를 免疫原으로 하여 BABL/c 생쥐로부터 單一클론抗體를 생산하고 그 특성을 구명하였던 바 다음과 같은 결과를 얻었다. 1) 3마리의 BABL/c 생쥐(YⅠ, YⅡ, ZI)에 돼지卵子의 透明帶를 免疫化하고, 複合抗體 生成을 확인한 후 생쥐의 脾臟細胞와 Myeloma(SP2/O-Ag14)를 polyethylene glycol를 融合을 실시한 결과 각각 25.8%, 54.5% 그리고 59.7%의 融合效率을 나타내었으며, ELISQ에 의해 陽性反應을 조사한 결과 각각 17.3%, 32.6% 그리고 6.2% 陽性反應 效率을 나타내었다. 2) YI에서 강한 陽性反應을 보인 6개의 well에 대한 cloning을 실시하고 抗體檢證을 행한 결과 20.8% ∼ 48.4%의 Cloning效率과 54.6% ∼ 82%의 陽性反應 效率을 나타내었다. 3) 강한 陽性反應을 나타낸 항체에 대해 sandwich ELISA法에 의해 isotype을 決定하였던바 2E93C(YⅠ), 3E83E7(YⅠ), 4E3(YⅡ)각각 IgG₂b, IgG₂a, IgM으로 확인되었다. 4) Isotype이 決定된 2E93C9(YⅠ), 3E84E7(YⅠ), 4E3(YⅡ)의 세포를 생쥐의 腹腔에 주사하여 얻은 腹水를 indirect ELISA法에 의해 titer를 決定한 결과 모두 1:400,000 이상의 높은 titer를 나타내었다. 5) 處理區로서 單一클론抗體의 腹水와 對照區로서 normal mouse serum이 각각 2%씩 함유된 배양액속에서 난자를 배양한 후 顯徵鏡下에서 관찰했을 때 對照區에서 배양된 난자의 표면은 정상적인 형태를 나타냈으나 處理區에서 배양된 卵子는 표면에 뚜렷한 沈澱層을 형성하였다. 6) 處理區와 對照區 卵子를 Rabbit anti-mouse IgG-FITC가 1% 함유된 배양액속에서 배양하고 洗滌한 후 螢光顯徵鏡下에서 관찰한 바 處理區의 卵子는 透明帶 주위에서 螢光이 나타났으나, 對照區에서는 나타나지 않았다. This study was carried out ot produce and characterize monoclonal antibodies against porcine zona pellucida, and undertaken as a basic study to develop immunocontraceptive vaccine and to investigate the function of zana pellucida in early fertilization process. The results obtained were summarized as follows: 1. Spleen cells of three BALB/C mice(YⅠ, YⅡ and ZI) which were immunized with porcine zona pelucida were fused with myeloma cells(SP2/O-Ag14) by polyethylene glycol. At the result of fusion, fusion efficiency was 25.8 , 54.5% and 59.7% and positive efficiency 17.3%, 32.6% and 6.2%, respectively. 2. Cloning efficiency was shown to be from 20.8% to 48.4% and positive efficiency of them were 54.6% to 82%. 3. Sub-isotypes of three strong positive antibodies, 2E93C(YⅠ), 3E83E7(YⅠ) and 4E3(YⅡ) were determined by sandwich ELISA method and shown to be as IgG2b, IgH2a and IgM, respectively. 4. The titers of three ascitic fluids containing antibodies, 2E93C9(YⅠ), 3E84E7(YⅠ) and 4E3(YⅡ) were determined by indirect ELISA and all of them showed above 1:400,000. 5. The layer of precipitate was observed on the zona incubated with medium containing 2% ascitic fluid of monoclonal antibody while the eggs treated with 2% normal mouse serum did not. 6. Porcine eggs incubated with medium containing 2% ascitic fluid of monoclonal antibody and followed by subsequent incubation with Rabbit anti-mouse IgG-FITC conjugate showed strong fluorescent light on the zona surface while the zona of normal mouse serum-treated eggs did not show fluorescence.

국내 감염성질환 병원체의 성상분석을 통한 표준실험실 체계 구축(Ⅱ) : 장내바이러스와 B형간염바이러스 국내 분리주의 성상분석(1)

윤재득,지영미,김기순,천두성,안정배,정윤석,신수연,이선화,이지원,조해월 국립보건연구원 2000 국립보건원보 Vol.37 No.-

200 GeV/핵자 유황이온과 핵건판핵의 충돌에 의해 생성된 헬륨 파쇄핵의 극한파쇄 연구

김동철,송진섭,윤천실,정성헌,박인곤,김종오,김철수,김태연,이승희,조재희,천병구,김재률,김준원,김태익,박명렬,장한일,임인택 慶尙大學校 기초과학연구소 1992 基礎科學硏究所報 Vol.8 No.-

고에너지 중이온 원자핵과 핵건판의 충돌에서, 200GeV/핵자 유황이온에 의해 생성된 파쇄 헬륨핵(Z=2)의 실험실계의 방출각 분포는 표적핵에 무관한 회귀공식. dN=exp[a+k exp(η-y_b)]d[exp(η-y_b)]로 잘 표현된다. 여기에서 의사신속도 η=-ln[tan(θ/2)]이고, y_b는 실험실계의 입사입자(^32S)의 신속도이다. 이 공식에 의한 적합에서 k=-0.057±0.008로 얻어진다. 즉, 핵건판과 고에너지 중이온의 충돌에서 파쇄 헬륨핵의 exp(η-y_b)의 분포는 "극한파쇄" 현상을 잘 설명하고 있다. The angular distribution of emission angle θ of helium (Z=2) produced in the collisions of incident particles of 200 GeV/nucleon ^32S in nuclear emulsion is well expressed by dN=exp[a+k exp(η-y_b)]d[exp(η-y_b)] where the pseudorapidity is η=-ln[tan(θ/2)], the laboratory system primary rapidity is y_b, and k=-0.057+0.008. The shape of this frequency of occurrence distributions in terms of exp(η-y_b) attests to the validity of the concept of "limiting fragmentation" for helium projectile fragments produced in the projectile fragmentation regions of heavy ion collisions in nuclear emulsion.

Clostridium difficile에 대한 분자역학 분석 방법 비교 연구

정경태,정예선,성원근,오희복 국립보건연구원 2000 국립보건원보 Vol.37 No.-

逆流浸降 電流法에 依한 돼지의 X-, Y-精子分離 關한 硏究

李用斌,呂政秀,鄭用基,尹種擇,徐國聖 서울大學校 農科大學 1980 서울대농학연구지 Vol.5 No.2

This experiment was carried out to separate X- and Y- bearing sperm of boar semen from 4 boars raising at the College of Agriculture, Seoul National University. The boar semen collected was treated by the counter streaming sedimentation and galvanization. The results obtained were as follows; Average appearance of B-body from control was 45.2%, when the experiment was performed with counter streaming sedimentation alone appearance of B-body from top and bottom fraction were 59.4% and 36.3%, respectively. The frequency of appearance of B-body from 30(24%) and 40(19.6%) minutes among treatments were statistically significant differences. Appearance of B-body from anode was 54% and from cathode was 37.9% after counter streaming sedimentation and galvanization. The difference between anode and cathode was significant differences at the 30(14.7%), 40(19.4%) and 50(19.9%) minutes. The frequency of appearance of B-body from counter streaming galvanization was significantly higher than that from counter streaming sedimentation. Therefore, the result from counter streaming sedimentation(30 minutes) and galvanization(30 minutes) was the most useful.

Influences of Enzyme Complex Supplementation on Growth, Ileal and Apparent Fecal Digestibility and Morphology of Small Intestine in Pigs

Kim, B.G.,Tian, J.Z.,Lim, J.S.,Kil, D.Y.,Jeon, H.Y.,Chung, Y.K.,Kim, Y.Y. Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.12

A total of 140 weaning pigs were used to determine the effects of digestive enzyme supplementation to corn-soybean meal diets on growth performance, physiological changes of small intestine, microorganisms and pH in the gastrointestinal tract. Two kinds of enzyme complex (A, B) were used in this experiment. Pigs were allotted in a completely random design (CRD) to five replicates with four pigs per pen. Diets and water were provided for ad libitum consumption. Treatments included 1) Control: without enzyme supplementation, 2) Enzyme A 0.05%, 3) Enzyme A 0.10%, 4) Enzyme A 0.15%, 5) Enzyme B 0.05%, 6) Enzyme B 0.10%, 7) Enzyme B 0.15% in the diets. A total of 24 crossbred barrows 25.78${\pm}$0.55 kg BW fitted with simple ileal T-cannulas were used to evaluate the effect the enzyme addition on the nutrient digstibility. Pigs were allotted 4 treatments (No enzyme, enzyme A 0.05%, enzyme A 0.1%, enzyme A 0.15%), 6 replicates according to a completely random design (CRD). Another digestibility trial was followed for enzyme complex B. Twenty pigs, average 31.92${\pm}$0.37 kg BW, fitted with simple ileal T-cannulas for digestibility trial. Neither enzyme A nor enzyme B affected on fecal or ileal digestibility of dry matter, gross energy, crude protein, crude fat and crude ash (p>0.05). The apparent fecal digestibilities of all the nutrients were higher in total feces collection method than in indirect method. At the end of feeding trial, 21 pigs were slaughtered for examining the morphological changes of small intestine and the concentration of microorganisms in the ileum and the colon. Growth performance, intestinal morphology and pH of ileum and colon were not affected by the either enzyme complex supplementation (p>0.05). These results suggested that enzyme complex A and enzyme complex B were of no benefit to early-weaned pigs when corn-soybean meal based diet was provided.

Bacteroides fragilis의 독소 유전자 분석 및 발병기전에 관한 연구

오희복,이근영,정경태,성원근 국립보건연구원 2000 국립보건원보 Vol.37 No.-

Clostridium difficile에 대한 분자역학적 Database 구축

오희복,정경태,정예선,성원근 국립보건연구원 1999 국립보건원보 Vol.36 No.-

Inhibition of NF-κB-induced inflammatory responses by angiotensin II antagonists in aged rat kidney

Kim, J.M.,Heo, H.S.,Choi, Y.J.,Ye, B.H.,Mi Ha, Y.,Seo, A.Y.,Yu, B.P.,Leeuwenburgh, C.,Chung, H.Y.,Carter, C.S. Pergamon Press ; Elsevier Science Ltd 2011 Experimental Gerontology Vol.46 No.7

In this study, we explored the mechanisms by which the angiotensin converting enzyme inhibitor (ACEI), enalapril, and the Ang II receptor blocker (ARB), losartan suppress oxidative stress and NF-κB activation-induced inflammatory responses in aged rat kidney. The experimentations were carried out utilizing aged (24-month-old) Brown NorwayxFischer 344 (F1) male rats which were randomized into 3 groups and administered enalapril (40mg/kg), losartan (30mg/kg) or placebo for 6months (daily p.o.). The level of reactive species (RS), peroxynitrite (ONOO<SUP>-</SUP>), GSH/GSSG and lipid peroxidation were measured. The activity of the pro-inflammatory transcription factor NF-κB, and gene expression of proteins in upstream signaling cascades were measured by electro-mobility shift assay (EMSA) and Western blotting. Enalapril and losartan differentially attenuated redox imbalance and the redox-sensitive transcription factor, the NF-κB pathway. Furthermore, stimulation of the NF-κB activation pathway by phosphorylation of p65 was attenuated by both compounds. Moreover, mediation of phosphorylation of p65 by phosphorylation of IκB kinase αβ (IKKαβ) and mitogen- and stress-activated protein kinase-1 (MSK-1), were also inhibited by enalapril and losartan. Finally, both compounds also lowered expression of NF-κB-dependent inflammatory genes, such as cyclooxygenase-2 (COX-2), and inducible NO synthase (iNOS). Only losartan lowered levels of 5-lipoxygenase (5-LOX). These findings indicate that enalapril and losartan differentially suppress inflammatory responses via inhibition of oxidative stress-induced NF-κB activation in aged rat kidney.

전기영동과 산알카리처리 결합방법에 의한 돼지정액의 X 와 Y 정자 분리에 관한 연구

이용빈,서국성,여정수,정용기 ( Y . B . Lee,G . S . Suh,J . S . Yeo,Y . K . Chung ) 한국축산학회 1980 한국축산학회지 Vol.22 No.4

The combination of the acid-alkali method and eletrophoresis for X-and Y-sperm separation of boar semen was performed We used lactic acid and sodium bicarbonate in acid-alkali method, and electrophoresed at 2.5V and 200 μA. The results obtained were summarized as follows : 1. The frequency of appearance of B-body from alkali treatment of whole semen was significantly higher than that from acid treatment. 2. The frequency of appearance of B-body from alkali treatment at the anode electrode was significantly higher compared with that from acid treatment at the same electrode, while that from acid treatment at the cathode was slightly higher than that from alkali treatment at the same electrode. 3. According to time of treatment, Appearance of B-body from alkali treatment at the anode after electrophonesis was decreased, and the result from acid treatment at the same condition was the opposite, while appearance of B-body from alkali treatment at the cathode after electrophoresis was not changed and that from acid treatment at the same condition was increased. 4. Appearance of B-body from alkali treatment at the anode for 5-10minutes showed the most useful result which was higher than the result (51.9%) from electrophoresis alone by 3 %.