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.

Aquaporin 2-labeled cells differentiate to intercalated cells in response to potassium depletion

Kim, W. Y.,Nam, S. A.,Choi, A.,Kim, Y. M.,Park, S. H.,Kim, Y. K.,Kim, J. Springer Science + Business Media 2016 Histochemistry and cell biology Vol.145 No.1

<P>The mammalian renal collecting duct consists of principal cells (PCs) and intercalated cells (ICs). Both PCs and ICs are involved in potassium (K+) homeostasis, PCs through their role in K+ secretion and ICs through their ability to facilitate K+ resorption. We previously hypothesized that PCs may differentiate into ICs upon K+ depletion. However, no direct evidence has yet been obtained to conclusively demonstrate that PCs differentiate into ICs in response to K+ depletion. Here, we present direct evidence for the differentiation of PCs into ICs by cell lineage tracing using aquaporin 2 (AQP2)-Cre mice and R26R-EYFP transgenic mice. In control mice, AQP2-EYFP+ cells exhibited mainly a PC phenotype (AQP2-positive/H+-ATPase-negative). Interestingly, some AQP2-EYFP+ cells exhibited an IC phenotype (H+-ATPase-positive/AQP2-negative); these cells accounted for 1.7 %. After K+ depletion, the proportion of AQP2-EYFP+ cells with an IC phenotype was increased to 4.1 %. Furthermore, some AQP2-EYFP+ cells exhibited a 'null cell' phenotype (AQP2-negative/H+-ATPase-negative) after K+ depletion. Collectively, our data demonstrate that AQP2-labeled cells can differentiate into ICs, as well as null cells, in response to K+ depletion. This finding indicates that some of AQP2-labeled cells possess properties of progenitor cells and that they can differentiate into ICs in the adult mouse kidney.</P>

식물성 세포벽에 함유된 Ca과 P의 Rooster에 있어서 소화율에 관한 연구

김종쾌,김영규,김대진 東亞大學校 1992 東亞論叢 Vol.29 No.1

본 연구는 綠葉중에서 ladino clover 잎, alfalfa 잎, 칡잎, 아카시아잎, 싸리잎 등을 5월과 6월의 生育시기를 두 시기로 하여 동물 체내에서 Ca과 P 그리고 NDF에 결합된 Ca과 P의 이용성을 조사하였다. 실험에 사용된 동물은 10개월된 재래 숫탉(rooster)으로서 이들 綠葉은 체중 kg당 약 1% 강제 급여(forced feeding) 한 후 36시간 동안의 배설물을 채취하였으며, 이와 동시에 대사성 糞과 내인성 뇨로 분비되는 Ca과 P을 보정하기 위하여 36시간 절식시켜서 배설물을 채취하였다. 생체내 Ca과 P의 이용성 실험은 세번 반복을 하였으며, 대사성 분과 내인성 뇨 실험도 다섯번 반복을 실시하였는데 그 결과를 요약하면 다음과 같다. 1. NDF 含量에 있어서 綠葉(ladino clover잎, alfalfa 잎, 칡잎, 아카시아잎, 싸리잎)은 5월 예취가 30.42%-37.28% 범위였으며, 6월 예취는 45.43-59.36% 점위로 生育이 進行됨에 따라 增加하였다. (p<0.05). 2. 眞正 DM 消化率(TDMD)에 있어서 綠葉은 5월 예취가 24.75-47.24%의 범위였으며, 6월 예취는 23.31-42.20% 범위로 生育이 進行됨에 따라 減少하였다(p<0.01). 3. 眞正 이용 Ca(TACa)에 있어서 綠葉은 5월 예취가 7.94-56.65%의 범위였으며, 6월 예취는 4.02-51.09% 범위로 生育이 進行됨에 따라 alfalfa 잎은 增加하였으나, ladino clover 잎은 減少하였고(p<0.05), 칡잎, 아카시아잎은 뚜렷한 변화가 없었다. 4. 眞正 이용 P(TAP)에 있어서 綠葉은 5월 예취가 8.52-4.89%의 범위였으며, 6월 예취는 24.86-42.6%로 生育이 進行됨에 따라 增加하였으나 ladino clover 잎에서는 減少하였다(p<0.05). 5. NDF에 결합된 Ca의 이용율에 있어서 綠葉은 5월 예취가 4.04-15.80%의 범위였으며, 6월 예취는 3.23-8.11%로 生育이 進行됨에 따라 減少하였으나 (p<0.05), ladino clover 잎, 싸리잎은 뜨렷한 변화가 없었다. 6. NDF에 결합된 P의 이용율에 있어서 綠葉은 5월 예취가 3.97-17.01% 범위였으며, 6월 예취는 4.06-14.93%로 生育이 進行됨에 따라 ladino clover잎, 칡잎, 아카시아잎은 減少하였으나(p<0.05), la-dinl clover 잎과 싸리잎은 뚜렷한 변화가 없었다. 7. 綠葉에 있어서 TDMD(Y) 含量과 NDF 含量(X) 사이에는 Y=53.5-0.42X의 回歸式과 r=-0.50의 相關關係를 보였다(p<0.01). 8. Ladinl clover 잎의 P-D(Y)와 P 含量(X) 사이에는 Y=84.5-85.1X의 回歸式과 r=-0.91의 相關關係를 보였다(p<0.01). 9. Alfalfa 잎의 Ca-D(Y)와 Ca含量(x) 사이에는 Y=18.9+13.0X의 回歸式과 r=0.99의 相關關係를 보였다(p<0.01). 10. 칡잎의 P-D(Y)와 P含量(X) 사이에는 Y=139-368X의 回歸式과 r=-0.95의 相關關係를 보였다(p<0.01). 11. 아카시아잎의 P-D(Y)와 P含量(X) 사이에는 Y=87.8-229X의 回歸式과 r=-0.98의 相關關係를 보였다(p<0.01). 12. 싸리잎의 Ca-D含量(Y)과 Ca含量(X) 사이에는 Y=17.9-12.8X의 回歸式과 r=-0.83의 相關關係를 보였다(p<0.01). This experiment was conducted to evaluate the Ca and P bioavailabilities of green leaf meal by roos-ter. The cell wall binding Ca and P of green leaf meal such as ladino clover, alfalfa, kudzu, acacia and bush clover were collected by two stage of cutting for May and June 1990. Rooster employed in the present experiment were fed about one percentage of body weight by forced feeding of ingredient. Collection of excreta as fed after 36 hous and did not given ingredient during the determinate of urinary endogenous and metabolic fecal Ca, P and excreta for 36 hours. The feeding experiment was conducted triplication for bioassay of Ca and P and was give five repli-cation for composition of metabolic and endogenous material. The results obtained are summarized as follows : 1. The contents of neutral detergent fiber(NDF) of several green leaves meal were ranged from 30.42% to 37.28% for cutting of May and were ranged from 45.43% to 59.36% for cutting of June, there-fore NDF were decreased by abvanced stages of maturity(p<0.05). 2. The true dry matter digestibility(TDMD) of several green leaves meal were ranged from 24.57% to 42.24% for cutting of May and were ranged from 23.31% to 42.20% for cutting of June and TDMD was decreased by advanced stages of maturity(p<0.05). 3. The true available clacium(TACa) of several green leaves meal were ranged from 7.94% to 56.65% for cutting of May and were ranged from 4.02% to 51.09% for cutting of June, exception of alfalfa among other did not show increase by advanced stages of maturity(p<0.05). 4. The true avilable phosphorus(TAP) of several green leaves meal were ranged from 8.25% to 46.89% for cutting of May and were ranged from 24.86% to 42.6% for cutting of June and TAP was increased but decreased only ladino clover by advanced stages of maturity(p<0.05). 5. NDF binding of avilable calcium(NDF+Ca) of several green leaves meal were ranged from 4.04% to 15.80% for cutting of May and were ranged from 3.23% to 50 8.11% for cutting of June and they were decreased by advanced stages of maturity(p<0.05) but ladino clover and bush clover did not changed by advanced stage of maturity. 6. NDF binding of avilable phosphorus(NDF+P) of several green leaves meal were ranged from 3.97% to 17.01% for cutting of May and were ranged from 4.06% to 14.93% for cutting of June and they were decreased by advanced stages of maturity(p<0.05) but alfalfa and bush clover did not changed by advanced stage of maturity. 7. Regression equation and correlation coefficients of several green leaves meal between TDMD(Y) and NDF(X) were Y=53.5=0.42X and r=-0.50(p<0.01). 8. Regression equation and correlation coefficients of ladino clover leaf meal between digestible phos-phorus(Y) and phosphorus(X) were Y=84.5-85.1X and r=-0.91(p<0.01). 10. Regression euqation and correlation coefficients of kudzu leaf meal between digestible phosphours(Y) and phosphours(X) were Y=139-368X and r=-0.95(p<0.01). 11. Regression euqation and correlation coefficients of acacia leaf meal between digestible phosphours(Y) and phosphours(X) were Y=87.8-229X and r=-0.98(p<0.01). 12. Regression euqation and correlation coefficients of bush clover leaf meal between digestible calcium(Y) and calcium(X) were Y=17.9-12.8X and r=-0.83(p<0.05).

Cellulase-amyloglucosidase와 효모의 가스생성법에 의한 사료의 에너지가 측정에 관한 연구

김영길,이광목,김욱 東亞大學校生命資源科學大學附設 農業資源硏究所 1993 農業生命資援硏究 Vol.2 No.2

粗飼料와 濃厚飼料를 混合한 飼料의 營養價를 Cellulase와 Amyloglucosidase, 酵母를 利用한 가스 生成法에 의하여 消化率과 TDN을 測定할 수 있는 方法을 確立하고자 本 硏究를 實施한 實驗 結果는 다음과 같다. 1. Cellulose 加水分解 酵素인 Trichoderma viride cellulase의 最適 活性 pH는 4.5-4.8이었으며 pH가 5.0보다 높을 때에는 活性이 急激히 저하하였다. 2. Cellulose 加水分解시 Trichoderma viride celulase의 溫度의 의한 影響은 50℃에서 最大 活性을 나타내었으며, 40℃에서는 活性이 92%정도로 나타났다. 3. Trichoderma viride-aspergillus niger 混合, cellulase 使用時의 가스 生成量은 Trichoderma viride cellulase 使用時 가스 生成量 보다 20-30% 높게 나타났으므로 cellulose 加水分解時 Trichoderma viride-aspergillus niger 混合 cellulase를 使用하는 것이 더욱 效果的일 것으로 思料된다. 4. 포도당(X)과 가스 生成量(Y)과의 關係는 Y=6.82+165X의 回歸式과 相關係數는 0.92로서 高度의 有意性(P<0.01)이 있었다. 5. Trichoderma viride cellulase 使用時 가스 生成量(X)과 乾物 消化率(Y)과의 關係는 Y=29.9+1.13X(r=78**)의 回歸式을 나타내었고, Trichoderma viride-aspergillus niger 混合 vellulase 使用時 가스 生成量(X)과 乾物 消化率(Y)과의 關係는 Y=28.9+0.88X(r=0.82**)의 回歸式을 나타내었다. Trichoderma viride cellulase 使用時 보다 Trichoderma viride-aspergillus niger 혼합 cellulase 사용시 相關係數가 더 높았다. 6. 가스 生成量(X)과 TDN(Y)과의 關係는 Y=35.8+0.585X의 回歸式과 相關係數 0.93으로서 高度의 有意性을 나타내었고, TDN(X)과 가스 生成量(Y)과의 關係는 Y=1.68X-59.6의 回歸式을 나타내었다. The experiment was conducted to establish the gas production method by cellulase, amyloglucosidase and yeast to determine the digestibility and energy value(TDN) of feedstuffs. The results obtained were summarized as follows; 1. The highest activity of Trichoderma viride cellulase was shown at pH 4.8 and the activity of the cellulase decreased rapidly at higher pH than 5.0. 2. The highest activity of Trichoderma virie cellulase was shown at 50℃ and the activity at 40℃ was approximately 92% compared with that at 50℃. 3. The mixed celllulases of Trichoderma viride and Aspergillus nigher produced more gas than Trichoderma viride cellulase when incubated with feedstuffs and were identified to be more effectively used for gas production method. 4. The regression equation of gas production(Y) on glucose production (X) was Y=6.82+165X with significant correlation coefficient of 0.92. 5. The regression equation of DMD(Y) on gas production(X) was Y=29.9+1.13X(r=0.78) with Trichoderma viride cellulase and Y=28.9+0.88X (r=0.82) with both Trichoderma viride and Aspergillus niger cellulase. 6. The regression equation of TDN(Y) on gas production(X) was Y=35.8+0.58X with significant correlation coefficients of 0.93 and in conclusion energy value of feedstuffs can be estimated by gas production method.

The partitioned-layer index: Answering monotone top-k queries using the convex skyline and partitioning-merging technique

Heo, J.S.,Whang, K.Y.,Kim, M.S.,Kim, Y.R.,Song, I.Y. North-Holland [etc ; Elsevier Science Ltd 2009 Information sciences Vol.179 No.19

A top-k query returns k tuples with the highest (or the lowest) scores from a relation. The score is computed by combining the values of one or more attributes. We focus on top-k queries having monotone linear score functions. Layer-based methods are well-known techniques for top-k query processing. These methods construct a database as a single list of layers. Here, the ith layer has the tuples that can be the top-i tuple. Thus, these methods answer top-k queries by reading at most k layers. Query performance, however, is poor when the number of tuples in each layer (simply, the layer size) is large. In this paper, we propose a new layer-ordering method, called the Partitioned-Layer Index (simply, the PL Index), that significantly improves query performance by reducing the layer size. The PL Index uses the notion of partitioning, which constructs a database as multiple sublayer lists instead of a single layer list subsequently reducing the layer size. The PL Index also uses the convex skyline, which is a subset of the skyline, to construct a sublayer to further reduce the layer size. The PL Index has the following desired properties. The query performance of the PL Index is quite insensitive to the weights of attributes (called the preference vector) of the score function and is approximately linear in the value of k. The PL Index is capable of tuning query performance for the most frequently used value of k by controlling the number of sublayer lists. Experimental results using synthetic and real data sets show that the query performance of the PL Index significantly outperforms existing methods except for small values of k (say, k=<9).

MOBILE NYLON BAG 기법에 의한 NaOH 처리 보리의 젖소 각 소화관내 영양소 분해율 측정

김병기(B . K . Kim),문여황(Y . H . Moon),이상철(S . C . Lee) 한국영양사료학회 1995 韓國營養飼料學會誌 Vol.19 No.4

Proposed mechanism in the change of cellular composition in the outer medullary collecting duct during potassium homeostasis.

Park, E-Y,Kim, W-Y,Kim, Y-M,Lee, J-H,Han, K-H,Weiner, I D,Kim, J Gutenberg 2012 HISTOLOGY AND HISTOPATHOLOGY Vol.27 No.12

<P>Potassium depletion (K?-D) induces hypertrophy and hyperplasia of collecting duct cells, and potassium repletion (K?-R) induces regression of these changes. The purpose of this study was to examine the time courses of the changes in cellular composition, the origin of intercalated cells (ICs) and the mechanism responsible for these changes. SD rats received K?-depleted diets for 1, 7, or 14 days. After K?-D for 14 days some of the rats received normal diets for 1, 3, 5, or 7 days. In the inner stripe of the outer medulla, K?-D increased significantly the number and proportion of H?-ATPase-positive ICs, but decreased the proportion of H?-ATPase-negative principal cells (PCs). However, proliferation was limited to H?-ATPase-negative PCs. During K?-R, the cellular composition was recovered to control level. Apoptosis increased during K?-R and exclusively limited in H?-ATPase-negative PCs. Double immunolabeling with antibodies to PC and IC markers identified both cells negative or positive for all markers during both K?-D and K?-R. Electron microscopic observation showed that ultrastructure of AE1-positive some cells were similar to AE1-negative some cells during K?-R. LC3 protein expression increased significantly and autophagic vacuoles appeared particularly in PCs on days 14 of K?-D and in ICs on days 3 of K?-R. These results suggest that PCs and ICs may interconvert in response to changes in dietary K+ availability and that autophagic pathways may be involved in the interconversion.</P>

Pepsin - Cellulase 에 의한 국내산 주요 조사료의 DMD 에 관한 연구 (3) 주요목초의 세포벽 구성물질과 건물소화율 (乾物消化率)

김대진(D . J . Kim),김영길(Y . K . Kim),황태기(T . K . Hwang),맹원재(W . J . Maeng) 한국영양사료학회 1989 韓國營養飼料學會誌 Vol.13 No.4

칡의 대량생산 이용과 생리활성물질에 관한 연구 (2) 당밀 , 칡즙 및 칡뿌리의 첨가가 칡사일리지의 발효품질과 사료적 가치에 미치는 영향

김대진(D . J . Kim),김영규(Y . K . Kim),맹원재(W . J . Maeng) 한국영양사료학회 1994 韓國營養飼料學會誌 Vol.18 No.2

代數的 擴大體에 關한 硏究

金勇基 한국항공대학교 1979 論文集 Vol.15 No.-

體 F가 體 K의 部分體일 때 K를 F의 擴大體라 한다. K의 한 元素를 C라 할때 ?? 을 滿足하는 ?? (모두는 0이 아님)가 F內에 存在할 때 C는 F 上의 代數的 元이 된다. aεK인 a가 F 上의 代數的 元일 때에 限하여 F(a)는 F의 有限擴大體가 됨을 證明하였다. 더 나아가서 L이 K의 代數的 擴大體이면 L은 F의 代數的 擴大體임을 밝혔다. When a field F is a subfield of a field K, K is called the extension field of F. If the field F has α?? satisfying the condition that ?? for an element C of K, C can be an algebraic element on the field F. And only when an element C of K is an algebraic element on F, F(a) becomes a finite extension field of K. Furthermore, in case L is an algebraic extension field of K, and K is that of F, L becomes an algebraic extension field of F.