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.

Long-term outdoor cultivation by perfusing spent medium for biodiesel production from Chlorella minutissima

Oh, S.H.,Kwon, M.C.,Choi, W.Y.,Seo, Y.C.,Kim, G.B.,Kang, D.H.,Lee, S.Y.,Lee, H.Y. Society for Bioscience and Bioengineering, Japan ; 2010 Journal of bioscience and bioengineering Vol.110 No.2

A unique perfusion process was developed to maintain high concentrations of marine alga, Chlorella minutissima. This method is based on recycling cells by continuous feeding with warm spent sea water from nuclear power plants, which has very similar properties as sea water. A temperature of at least 30 <SUP>o</SUP>C in a 200 L photo-bioreactor was maintained in this system by perfusion of the thermal plume for 80 days in the coldest season. The maximum cell concentration and total lipid content was 8.3 g-dry wt./L and 23.2 %, w/w, respectively, under mixotrophic conditions. Lipid production was found to be due to a partially or non-growth related process, which implies that large amounts of biomass are needed for a high accumulation of lipids within the cells. At perfusion rates greater than 1.5 L/h, the temperature of the medium inside the reactor was around 30 <SUP>o</SUP>C, which was optimal for cell growth. For this system, a perfusion rate of 2.8 L/h was determined to be optimal for maintaining rapid cell growth and lipid production during outdoor cultivation. It was absolutely necessary to maintain the appropriate perfusion rate so that the medium temperature was optimal for cell growth. In addition, the lipids produced using this process were shown to be feasible for biodiesel production since the lipid composition of C. minutissima grown under these conditions consisted of 17 % (w/w) of C<SUB>16</SUB> and 47% (w/w) of C<SUB>18</SUB>. The combined results of this study clearly demonstrated that the discharged energy of the thermal plume could be reused to cultivate marine alga by maintaining a relatively constant temperature in an outdoor photo-bioreactor without the need for supplying any extra energy, which could allow for cheap production of biodiesel from waste energy.

Validation of egg yolk antibody based C-ELISA for avian influenza surveillance in breeder duck

Jeong, O.M.,Kim, M.C.,Kang, H.M.,Ha, G.W.,Oh, J.S.,Yoo, J.E.,Park, C.H.,Kwon, J.S.,Pack, M.R.,Kim, H.R.,Kim, Y.J.,Kwon, J.H.,Lee, Y.J. Elsevier Scientific Pub. Co 2010 Veterinary microbiology Vol.144 No.3

Active surveillance for avian influenza virus (AIV) has expanded from chicken to various poultry species including duck. To further effective antibody screening in laying breeder ducks, we validated the egg yolk antibody as alternative source to serum for AIV antibody. Sera and eggs were collected at weekly intervals after two types of AIV vaccination, H5N3 and H9N2. The antibody levels were determined by an agar gel immunodiffusion (AGID) test, haemagglutination inhibition (HI) test and the competitive enzyme-linked immunosorbent assay (C-ELISA). AGID test did not detect antibodies in egg yolk, and the agreement between AGID test and either HI test or C-ELISA in serum was slight and fair based on kappa statistics (kappa value (κ)@?0.19 in H5N3 group and κ@?0.37 in H9N2 groups). However, there was almost perfect agreement between HI test and C-ELISA (κ>0.9 in all group). The C-ELISA was as sensitive and specific as the HI test, and could be used as a pre-screening test for the detection of type A avian influenza virus antibody. Comparison was made between egg yolk and serum antibody titers by a regression analysis. A high correlation was observed between serum and yolk antibody titers (r=0.8762 for H5N3 and 0.8914 for H9N2 in HI test; r=1 for H5N3 and 0.9686 for H9N2 in ELISA test), although egg yolk antibodies were detected later and remained lower levels than serum antibodies. In field trials involving 54 duck flocks, the positive rate of egg yolk and serum samples showed agreement for the detection of AIV antibody. We concluded that as an alternative to serum, antibody monitoring of laying breeder duck using egg yolk with C-ELISA is feasible and is recommended.

Single-tube nested PCR assay for the detection of avian botulism in cecal contents of chickens

Jang, I.,Lee, J.I.,Kwon, Y.K.,Kang, M.S.,Kim, H.R.,Park, J.Y.,Lee, S.H.,Lee, H.S.,Bae, Y.C. Academic Press 2015 Anaerobe Vol.35 No.2

This paper describes a novel diagnostic method for the detection of avian botulism caused by Clostridium botulinum type C and C/D, using single-tube nested PCR assay. This assay was developed to overcome the disadvantages of bioassays used in experiments with mice. Three primer pairs including an antisense primer were designed to target the N-terminal of the toxin gene from C. botulinum types C and C/D. The specificity of the PCR assay was confirmed by using 33 bacterial strains and chicken cecal contents from farms that experienced botulism outbreaks. The detection limit for purified DNA was 1.1 fg/μl, and for bacterial spores was 4.3 spores/200 mg of cecal contents. While checking for specificity of the PCR assay, the reactions with the templates form C. botulinum type C and C/D which were tested became positive, but the rest of the reactions turned negative. However, the results for all clinical samples (n = 8) were positive. The PCR assay results for cecal samples obtained from 300 healthy chickens (150 Korean native chickens and 150 broilers) were all negative. This assay is rapid and straightforward and evades ethical issues associated with mouse bioassay. Moreover, it is more economical than real-time PCR.

Effects of Dietary Fermented Chlorella vulgaris (CBT^®) on Growth Performance, Relative Organ Weights, Cecal Microflora, Tibia Bone Characteristics, and Meat Qualities in Pekin Ducks

Oh, S.T.,Zheng, L.,Kwon, H.J.,Choo, Y.K.,Lee, K.W.,Kang, C.W.,An, Byoung-Ki Asian Australasian Association of Animal Productio 2015 Animal Bioscience Vol.28 No.1

Fermented Chlorella vulgaris was examined for its effects on growth performance, cecal microflora, tibia bone strength, and meat qualities in commercial Pekin ducks. A total of three hundred, day-old male Pekin ducks were divided into three groups with five replicates (n = 20 ducklings per replicate) and offered diets supplemented with commercial fermented C. vulgaris (CBT$^{(R)}$) at the level of 0, 1,000 or 2,000 mg/kg, respectively for 6 wks. The final body weight was linearly (p = 0.001) increased as the addition of fermented C. vulgaris into diets increased. Similarly, dietary C. vulgaris linearly increased body weight gain (p = 0.001) and feed intake (p = 0.001) especially at the later days of the feeding trial. However, there was no C. vulgaris effect on feed efficiency. Relative weights of liver were significantly lowered by dietary fermented C. vulgaris (linear effect at p = 0.044). Dietary fermented C. vulgaris did not affect total microbes, lactic acid bacteria, and coliforms in cecal contents. Finally, meat quality parameters such as meat color (i.e., yellowness), shear force, pH, or water holding capacity were altered by adding fermented C. vulgaris into the diet. In our knowledge, this is the first report to show that dietary fermented C. vulgaris enhanced meat qualities of duck meats. In conclusion, our study indicates that dietary fermented C. vulgaris exerted benefits on productivity and can be employed as a novel, nutrition-based strategy to produce value-added duck meats.

C1~C3 급 볼스크류용 소재 국산화 개발

권정흠(J. H. Kwon),배완석(W. S. Bae),박춘영(C. Y. Park) 한국정밀공학회 2016 한국정밀공학회 학술발표대회 논문집 Vol.2016 No.10월

Loganin improves learning and memory impairments induced by scopolamine in mice

Kwon, S.H.,Kim, H.C.,Lee, S.Y.,Jang, C.G. North-Holland ; Elsevier Science Ltd 2009 european journal of pharmacology Vol.619 No.1

Loganin is an iridoid glycoside found in the Flos lonicerae, Fruit cornus, and Strychonos nux vomica. We investigated the effect of loganin on learning and memory impairments induced by scopolamine (0.5mg/kg, i.p.), a muscarinic antagonist, using the Y-maze, passive avoidance, and the Morris water maze tests in mice. In the Y-maze test, loganin (40mg/kg, p.o.) significantly improved the scopolamine-induced memory impairment. In addition, loganin (20 and 40mg/kg, p.o.) significantly reversed scopolamine-induced impairments measured by the passive avoidance and the Morris water maze tests. A day after the last trial session of the Morris water maze test (probe trial session), loganin (20 and 40mg/kg) dose-dependently increased the latency time in the target quadrant. Furthermore, loganin significantly inhibited acetylcholinesterase activity in the hippocampus and frontal cortex. Loganin may have anti-amnesic activity that may hold significant therapeutic value in alleviating certain memory impairments observed in Alzheimer's disease.

Genetic diversity and structure in apple-infesting pests of Carposina sasakii, Grapholita dimorpha and Grapholita molesta in Korea

Kwon, D.H.,Kim, D.H.,Kim, H.H.,Lee, S.H.,Yang, C.Y. 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol. No.

<P>The genetic diversity of three apple-infesting pests, Carposina sasakii, Grapholita dimorpha and Grapholita molesta, found in four local regions in Korea, was investigated using the mitochondrial cytochrome c oxidase I (COI) gene as a marker. Six, seven, and three larval haplotypes from C. sasakii, G. dimorpha, and G. molesta, respectively, were observed. The haplotype and nucleotide diversity were ranged as 0.31-0.69 and 0.0006-0.0040, respectively. The estimates of genetic differentiation (F-ST) among local populations in C. sasakii and G. dimorpha exhibited very narrow range differentiation as 0 to 0.025 and 0 to 0.075, respectively, whereas G. molesta revealed a moderate range of genetic distance (F-ST = 0-0.461). In the AMOVA analysis, low percent variations (0-0.98%) and F-ST , values (0 and 0.01, respectively) observed in C. sasakii and G. dimorpha, respectively, whereas G. molesta revealed moderate diversity as 31.3% (F-ST = 0313, p = 0.038). These results would provide the basic information to understand the difference of genetic structure in species level and further be utilized to estimate the temporal and spatial distribution of these species in apple orchard. (C) 2016 Korean Society of Applied Entomology, Taiwan Entomological Society and Malaysian Plant Protection Society. Published by Elsevier B.V. All rights reserved.</P>

갱년기 남성의 체험

권인영・김채연・김하정・신유정・이도경・장환희・정예은,김채연,김하정,신유정,이도경,장환희,정예은,양숙자,손지희 이화여자대학교 간호학회 2018 이화간호학회지 Vol.- No.52

Purpose: The purpose of this study is to obtain comprehensive understanding of Korean middle-age men's experience in their andropause. The findings of this study will provide supportive data for development of nursing interventions. Methods: This study is explored by hermeneutic phenomenological analytic method suggested by van Manen. Data was collected through in-depth interviews with 7 middle aged men between July and September, 2017. Before the interviews, preliminary questionnaire was preformed for all participants. Results: The analysis produced five themes: 'My body is getting weaker than before', 'A sense of despondency with current life', 'Regarding andropause not as much important as menopause', 'The obscurity of one's life about andropause', 'Cut off andropause from life'. Conclusion: This study explored Korean middle-age men's experience in their andropause. Although most participants experienced physical and psychological symptoms, they denied that they are andropause. Moreover, they lacked knowledge and showed attitude of indifference. In addition, Andropause is not as commonly recognized in the society of Korea. Even if they perceive it, they cannot reveal the fact and respond poorly. It brings up the necessity to provide proper information and education. A variety of nursing interventions are also required for the men for their physical or mental recovery and health care. Therefore, this study offers the deep understanding of the significance and nature of the experience in andropause. Furthermore, this study may put foundation for development in nursing care of studies of this subject.

Synthesis of anionic multichain type surfactant and its effect on methane gas hydrate formation

Kwon, Y.A.,Park, J.M.,Jeong, K.E.,Kim, C.U.,Kim, T.W.,Chae, H.J.,Jeong, S.Y.,Yim, J.H.,Park, Y.K.,Lee, J.d. Korean Society of Industrial and Engineering Chemi 2011 Journal of industrial and engineering chemistry Vol.17 No.1

This paper reports an experimental study on the effects of novel anionic multichain disulfonate surfactant on the formation of methane gas hydrate. A series of surfactants with sodium sulfonic acid groups and different hydrophobic carbon chain lengths (C<SUB>8</SUB>, C<SUB>10</SUB>, and C<SUB>12</SUB>) were synthesized, and the effects of carbon chain length and concentration on methane hydrate formation kinetics were systematically investigated. Methane hydrate formation was conducted in a magnetic stirred vessel at the constant temperature of 274.15K and in a pressure range of 3.5-4.0MPa. All surfactants showed kinetic promoting behavior for methane gas hydrate formation, and the surfactant with the shortest chain length showed the highest acceleration effect. In addition, this multichain disulfonate surfactant exhibited higher methane storage capacity than SDS (sodium dodecyl sulfate) even at lower surfactant concentration, due to its lower critical micelle concentration (CMC) and the surface tension itself of the Gemini-type multichain surfactant.