家兎繼代牛疫毒의 變異에 關한 硏究

李學喆 慶北大學校 1958 論文集 Vol.2 No.-

Nakamura and his associates reported that the pathogenesis of rabbit-passaged Strain Ⅲ of the Nakamura lapinized rinderpest virus (hereinafter referred to as the "Strain Ⅲ") is fixed in rabbit and cattle. However, we found that a serial passage of this "Strain Ⅲ" and its p thogenesis become more virulent and almost all the rabbits that were infected with this "Strain Ⅲ" died in 1947. If we assume that the increase in its virulence is due to the variation of its pathogenesis by the more serial passage of this "Strain Ⅲ", it is an importanr problem in the preventaion of epidemics of Rinderpest virus in cattle. An attempt was made in our laboratory to investigate the variation of the "Strain Ⅲ" and the following conclusions were made. 1) The increase of death rate in rabbit is not due to a mixinfection of other pathogenic organisms nor symbiosis with other organisms but it has been proved that it is due to the variation of its pathogenesis. 2) The pathologic change and the necrotic process of the lymph follicular cells in infected rabbits seem to be different in this study from the one reported previously by others. Particularly the cause of sever hemorrhagic change in the mucosa of gastrointestinal tract is not clear and more study will be needed to clearly its pathogenesis. 3) The minial infectious dose of this "Strain Ⅲ" in rabbits is higher in this study than in previous studies. 4) The neutralizaing potency of the convalescent serum of infected rabbit is more lower in this study than in previous works. 5) The following results were obtained by an inoculation of this "Strain Ⅲ" to Korean calves. a) The minimal infectious dose for calves is much higher in this study than in previous studies. b) There were died case and recovered case in claves inoculate with this "Strain Ⅲ". This is similar in this study to the previous works. c) Some difference was noted in appearance of critical symptoms in this study. d) The entire course and incubation period of infected calves were shorted considerably in this study than in previous works. 6) The results of simultaneous inoculation with this "Strain Ⅲ" plus antirinder pest hyperimmune serum to Korean calves also indicated that the pathogenesis of this "Strain Ⅲ" becaome more virulent than in previous studies. 7) The successive passage of theis "Strain Ⅲ" through Korean calves showed the following results. a) In calves the pathogenesis become more virulent but at no time it acquired the virulence of Rinderpest virus. b) The death rate of infected calves is higher in this study than in previous studies. c) When the blood of infected calves were inoculated into rabbits, fever and necrosis of the lymph follicular cells were seen and this characteristics of this "Strain Ⅲ" was maintained unitl the last passage of this study. d) Minimal infectious dose of blood from infected calves was markedly low ans this is due to the difficulty in reproducation of this "Strain Ⅲ" in calves. 8) The simultaneous inoculation with this "Strain Ⅲ" plus antirinderpest hype rimmune serum which is used in Korea is safe for cattles but it is dangerous for calves. For this reason increased dose of immune serum must be used for calves.

Newcastle病毒 接種雛의 眠房水로부터의 病毒分離에 對하여

李學喆,趙漢喆 慶北大學校 1962 論文集 Vol.6 No.-

Methods requiring virus isolation and confirmation of the virus as a means of diagnosing Newcastle disease (ND) are time consuming. Simple and rapid tests that have been proposed take advantage of the fact that ND virus belong to the myxovirus gropp and hemoagglutinates red blood cells. Clark et al. reported the use of the aqueous humor from eyes of chickens in the incipient stages of ND as a souce of antigen for HA testing. They indicate that the virus exists in this fluid before the blood serum inhibits the HA reaction. The same authors, after further studies with aqueous humor from asymptomatic vaccinated and unvaccinated chickens, concluded that the aqueous humor from asymptomatic vaccinated and unvaccinated chickens, concluded that the aqueous humor served as a reservior for ND virus. Dardiri et al., in contrast, could not demonstrate the virus in the aqueous humor of vaccinated chickens that survived challenge. Further study is needed to determine the persistence of the virus in the aqueous humor of convalesent and immune birds. This study, therefore, was conducted to investigate the persistence of ND virus In the aqueous humor of young chickens experimentally inoculated with the virulent strain (Kyojoungwon strain) and experimentally vaccinated with single dose of the B_1 living virus vaccine and challenged. The results are surmmarized as fellows : 1. ND virus was isolated from the aqueous humor both of vaccinated and unvaccinated chickens for the first about week following exposure to the virulent strin, however in previously vaccinated chickens, isolation of the challenged virus was sparse but in susceptible chickens, readily isolated from almost all of them except the few case. 2. Both of the vaccinated and unvaccinated chickens which took a prolonged course were not isolated readily.

Comparison of the Pharmacological Effects of Roasted and Non-roasted Zizyphi Spinosi Semen: Differences in Chemical Constituents

홍학기, 이기남, 마유안, 김윤배, 남상윤, 송인섭, 문동철, 오기완 충북대학교 동물의학연구소 2008 Journal of Biomedical and Translational Research Vol.9 No.3

본문참고

Wassermann反應의 變法에 關한 硏究 : 特히 Kolmer氏法에 對하여 Especially Kolmer's Method

李學喆,梁學道 慶北大學校 1962 論文集 Vol.5 No.-

The serum of the human suspected of being infected with Treponema pallidum has been tested by the Wassermann reaction, precipitation test (including flocculation test), Treponema pallidum agglutination test of Treponema pallidum immobilization test etc. The Wassermann reaction, however, has been found to be one of the most useful and reliable methods employed as laboratory aids to diagnosis. The methods of performing the test are legion and many modifications have been suggested to increase its reliability and sensitivity. Especially the cardiolipin-lecithin-cholesterol antigen investigated by Pangborn replaced effectively the crude tissue extract in most of current serodiagnostic test for syphilis. The authors undertook to disgnose a number of serum of the human suspected of being infected with syphilis in Pusan area, and the specimens were examined by Kolmer's method (serum dilution method) from the spring to the autumn of 1946. As a antigen the alcholic extract(added an alcholic solution of cholesterol) of heart-muscle from the healthy calf was employed in this examination. In performing the above procedure, the employed complement had to be treated with the cold method so as to be free of normal hemolysin in it, and varying amounts of the definite dilution of each of hemolysin, antigen and parient's serum were set up in a series of test tube, then they were titrated or examined with red blood cells and the other reagents respectively. Inasmuch as these amounts vary the volume of fluid in each tube was regulated by adding the amount of saline solution which brings the volume to 3㎖. or 4㎖. The various amounts are given in Table 1. In these respects the above method was extremely inconvenient, so that following modified method was made in our laboratory. The method is given in Table 3. 1. The amount of suspension of red blood cells was kept constant (2.5% 0.5㎖.) and the reagent to be titrated or examined in the preliminary test and complement fixation test was set up in a series of test tube with each 0.5㎖. of it diluted by two-fold series dilution(except the titration of complement), then each 0.5㎖. of saline solution was added to a series of test tube in order to bring the volume to 2.5㎖. 2. In the first place, the titration of hemolysin was tested with the complement including normal hemolysin and red blood cells, then the titration of complement was tested by using two units of the previously titrated hemolysin (included into 0.5㎖. of the definite dilution). And each two units of the titrated hemolysin and complement (included into 0.5㎖. of the definite dilution) were used in the following preliminary test (the titration of anticomplementary effect of antigen and antigenicity of antigen) and complement fixation test. 3. The complement fixation test was cardiod out with 0.5㎖. of the definite dilution of antigen determined by previous titraion and above each two units of hemolysin and complement. Repeated experiments with the above modified method have given consistently satisfactory results. The princeples of its procedure are in no way different from Kolmer's method and rationale of complement fixation in general.

大邱市內에 있어서의 山羊 및 緬羊의 結核症에 對한 調査

孫濟英,金容珌,李學喆 慶北大學校 1962 論文集 Vol.5 No.-

Tuberculosis is rare in sheep and goats. In sheep most of the infection that have been typed in the United States were found to be of the avian-type, but in orther countries most have proved to be of the bovine-type. The data on tuberculosis in goats are very meager, so this fact gives an impression on us that goats are relatively resistant to infection. Infections, however, with human-type, bovine-type and aviane-type have been rarely reported. The obtained specimens (lung, liver, spleen) from a died goat at Taegu Agriculture and Forestry High school in March 1960 were examined bacteriologically and pathologically to diagnose the cause of its death. Moreover, the 10 sheep and 13 goats which had been together in the same pen with the died goat and the 11 sheep and 202 goats which had been fed in the surrounding area of Taegu city were tested on the tuberculin reaction with the intradermal method. The tuberculin was mamalian type (O.T.). The results thus obtained would be outlined as follows: 1. The death of the goat at above school was caused with a generalized tuberculosis infected through respiratory route by the bovine-type of tubercle bacilli. 2. The result of tuberculin test have proved that among 10 sheep and 13 goats which were fed together with the died goat, the positive reaction was found in 2 sheep (20%) and 2 goats (about 15.4%) and the doubtful reaction was found in 1 sheep (10%) and 2 goats (about 15.4%), but among 11 sheep and 202 goats which were fed in the surrounding area of Taegu city, the positive and doubtful reaction were found in 2 goats (about 1%) and 3 goats (about 1.5%) respectively, and all of the sheep reacted in the negative.

Comparison of Immune Effect by Different Vaccination Route with Blacksburg Strain(B_1) Virous to Baby Chicks Hatched from Hens Immunized Against Newcastle Disease

Lee, Hak-Cheul,Son, Jae-Young 慶北大學校 1967 論文集 Vol.11 No.-

Newcastle病(ND)豫防液로서 Blacksburg Straiu(B_1) Virus를 우리 나라에導入한 以來 이 Virus를 應用한 幼雛免疫에 있어 몇가지의 報告가 있으나 모두 母鷄으로부터 이어받은 幼雛가 지니고 있는 先天的 免疫性을 考慮치 않은 것이었다. 著者는 앞서 그에 對한 基礎的 問題의 하나를 밝히기 위하여 母鷄로부터 初生雛에 移行된 ND, 血球凝集抑制(HI)抗體의 消長과 母鷄, 이에 由來한 鷄卵의 卵黃 및 初生雛間의 三者의 力價에 關해서도 比較檢討한 바가 있다. 이번의 硏究 目的은 上記知見을 基礎로하여 常規免疫鷄, 高度免疫鷄由來幼雛에 對하여 接種經路를 달리해서 B_1 Virus를 應用한 免疫成績에 關하여 比較觀察하였다. 이번 硏究에서 얻은 結論을 要約하면 다음과같다. 先天的免疫性을 지니고있는 幼雛들은 B_1 Virus 筋肉內接種에 對하여 妨害現象을 나타내어 免疫이 全혀 成立되지 않았으나 同量病毒을 鼻孔點適注入한 것에 있어서는 感染이 發展되었으며 攻擊毒의 筋肉內后毒接種에 對하여 어느 程度 低抗하였다. Chicks with congenital passive immunity appear refractory to intramuscular vaccination with Blacksburg strain(B_1) virus; however, the same chicks vaccinated with an identical dose of the same virus by nasal instillation seem to develop infection and subsequent immunity to intramuscular challenge exposure.

Expression of Speclfic Proteins in Hanwoo Cervical Mucus during Spontaneous and Induced Estrus Synchronization

Hak-Jae Chung,Nam-Kuk Kim,Hwi-Cheul Lee,Won-Kyong Chang,Su-Bong Park,Suk-Dong Lee,Dong-Min Yoo,Jin-Sung Ko,Yon-Ho Choy,Seong-Bok Choi,Gi-Jun Jeon,Myeoung-Sik Lee,Seok-Ki Im 한국동물번식학회 2009 Reproductive & Developmental Biology(Supplement) Vol.33 No.2s

Expression of Speclfic Proteins in Hanwoo Cervical Mucus during Spontaneous and Induced Estrus Synchronization

Hak-Jae Chung,Nam-Kuk Kim,Hwi-Cheul Lee,Won-Kyong Chang,Su-Bong Park,Suk-Dong Lee,Dong-Min Yoo,Jin-Sung Ko,Yon-Ho Choy,Seong-Bok Choi,Gi-Jun Jeon,Myeoung-Sik Lee,Seok-Ki Im 한국동물생명공학회(구 한국동물번식학회) 2009 Reproductive & developmental biology Vol.33 No.2

A Modified Procedure for the Assay of Fowl Leukosis Viruses by the Resistance Inducing Factor Tissue : Culture Method of Rubin

Lee, Hak-Cheul 慶北大學校 1965 論文集 Vol.9 No.-

腫瘍病毒(Tumor viruses)의 하나인 Rous Sarcoma virus (RSV)는 1910년 Rous에 依하여 처음으로 發見되였으며 그以後 많은 硏究者들에 依하여 광범위한 硏究가 施行되였다. 1941년 Halberstaedter등에 依하여 生體外 (invitro)에서 人工的으로 發育시킨 鷄纖維芽細胞에 本病毒을 感染시킬수 있다는 것과 1956年 Manaker 等에 依하여 鷄胎兒纖維芽細胞의 組織培養에 있어 RSV 感染으로 因한 病巢(Focus) 形成을 觀察할수 있다는 것이 報告된 以後 1958년 Temin 및 Rubin 等은 Dulbeco (1952)가 考案한 소위 Plaque 法(plaque method)를 利用하여 精密하고 信賴性 있는 生體外 RSV檢定에 關하여 報告하였다. 이 組織培養에서 볼수 있는 所謂 顯微的腫瘍(Microtumor) 形成는組織培養에 있어서의 各種因子로 因하여 安定치 않다는 것이 알려졌으나 1960年 Rubin은 이와 같은 모든 不利한 因子를 排除한 環境에서도 이 病巢形成이 Rubin이 命名한 Resistance inducing factor (RIF)에 依해서 적어도 정상보다 40배 以上이나 抑制 된다는 것을 報告하는 한편 이 因子는 一種의 病毒임을 推論하였다. 追後 이 現象의 本能는 病毒相互間에 惹起되는 干涉(Interference)에 起因된다는 것이 알려졌으며 Rubin이 命名한 RIF는 鷄白血病毒群(Avian leukosis viruses) 中의 內臟型淋巴腫症病毒(Visceral lymphomatosis virus)인 것이 여러 가지 面으로 立證되었다. 이와 같은 RSV를 가지고 하는 組織培養法은 追後 鷄白血病毒과 그의 抗體를 檢出하는 方法을 考案케 하였으나 實際面에 있어서 이들에 對한 具體的인 方法(Procedure)에 關하여는 Rubin 및 Temin 等의 文獻上에 詳細한 記載를 欠하고 單只 1962年 Piraino가 提案한것 以外로 同年 Solomon이 美農務省主催 鷄白血病 協議會時의 刊行物上에 若干 言及한 程度에 지나지 않다. 鷄白血病毒에 대한 組織培養法은 著者의 立場에서 볼때 일의 性質상으로 보아서 復雜性을 免치 못하는 그自體에 Piraino에 의하여 比較的 詳細히 提示된 方法은 너무나 復雜하게 構成되여 있음으로 著者는 이 方面에 關聯된 報告에 基礎를 두고 RSV, RIF, RSV抗體檢定(RIF 抗體檢定에 代置)等에 대해서 修正을 加하는 한편 著者 자신으로서의 術技를 樹立코저 硏究한 結果 病毒學적 立場에서 보와 合理的이라고 認定되는 이들에 對한 하나의 變法을 確立하였다. 이 報告에 記載된 內容은 1) 組織培養에 使用된 細胞의 出處 2) RSV에 依한 病巢形成 抑制因子 및 細胞發育 抑制因子 3) 組織培養에 使用된 器具, 容器, 病毒株, 培養液 및 溶液 4) 細胞의 培 養方法 5) 鷄胎兒纖維芽細胞의 準備 6) RSV 檢定法 7) 病巢의 計數法 8) RIF 對한 鷄胎兒 檢定法 9) RIF 檢定法 10) RSV抗體 檢定法 等에 對해서이며 本硏究에서 얻은 結果를 提示하고 考察을 加하였다. 이 硏究에서 얻은 結果와 結論을 要約하며 다음과 같다. 1. 一次培養(Primary culture)의 鷄胎兒纖維芽細胞는 著者의 方法에 依하여 準備되었다. 2. RIF에 對한 鷄胎兒檢定이 끝날때까지의 一次培養의 維持는 Piraino가 提案한 半流動體培地를 가지고 하는 方法代身에 液體養育培地를 가지고하는 方法을 應用함으로써 上記細胞培養을 8日間 以上이나 滿足히 維持함수 있었음. 3. RSV病毒에 對한 細胞의 感受性에 있어 15 l과 New York S系 鷄胎兒纖維芽細胞 사이에는 差異 가 없었음. 4. 犢血淸內에 存在하는 細胞發育抑制因子는 攝氏 56度 30分間 加熱함으로써 쉽게 破壞되였으다 . 이以外로 零下 攝氏 25度에서 一定동안 保存함으로써도 消滅되였으나 어떤 血淸은 되지 않았 다. 5. 犢血淸以外에 羊血淸을 鷄胎兒纖維芽細胞培養에 使用할수 있음을 認定하였으나 成牛血淸및 馬血淸은 不適이었다. 6. 犢血淸 使用에 있어 RSV 病毒으로 因한 病巢를 形成시킴에 對하여 障害가 없었음. 7. 鷄胎兒 纖維芽細胞發育에 對한 至適 水素lon濃度는 7.3이었다. 8. 培養細胞를 消化, 分散함에 있어 0.05% Trypsin 溶液을 使用하는 Rubin의 方法代身에 1 x ATV 溶液을 가지고 處理하는 方法을 適用하였다. 9. 一次培養은 RIF에 對한 鷄胎兒檢定이 끝날때 까지의 그의 細胞層 維持上으로 보아 慣例的 方 法에 依하여 培養하는 것이 可함. 10. 이 硏究에서 確立된 RSV檢定法은 本質的으로 보와서 以前에 報告된 原法에 類似하나 細胞培 養에 接種된 病毒量, 培養細胞에 病毒을 吸着시킴에 應用된 時間, 病毒接種後의 培養細胞의 取 扱 및 固形寒天培地를 가지고 培養細胞를 덮은 後의 細胞를 養育하는 方法등이 上記原法과 若干 다르다. 그리하여 이 RSV檢定에 應用된 方式은 RIF에 對한 鷄胎兒檢定, RIF檢定 및 RSV 抗體에 對한 檢定 등에 直接 連結되였다. 11. RIF感染細胞의 繼代에 있어 液體養育培地下에서 100萬個細胞를 繼代해나가면서 3日間隔을 두고 40萬個 細胞를 準備하여 3回에 걸천 Rous病毒攻擊을 滿足히 遂行할수 있었으나 同一條件下 에서 100萬個 혹은 200萬個細胞를 繼代하면서 上記와 같은 Rous病毒攻擊에 所要되는 100萬個細 胞의 繼代培養의 準備는 滿足히 遂行할수 없었음. 12. RSV抗體의 檢定에 있어 攝氏 37度 40分間 血淸과 病毒을 接觸시켜 病毒을 中和하는 方法 代 身에 攝氏 4度 18時間 接觸시키는 方法을 應用하였으다. 또한 可檢血淸의 稀釋은 5倍稀釋으로부 터 出發하였다. A procedure for the assay of avian leukosis viruses by the resistance inducing factor (RIF) tissue culture method of Rubin was studied to ascertain optimal conditions in the various factors concerning the method, and some modifications were developed for the assay of Rous sarcoma virus (RSV), visceral lymphomatosis virus (VLV) and antibody to RSV. The results and conclusions thus obtained would be outlined as follows: 1. The primary cultures of chicken embryo fibroblasts were prepared by the procedure developed by the author. 2. The liquid feed media system was applied instead of the soft agar modia system for the maintenance of primary cultures until the completion of the test of embryos for RIF activity. The cultures could be satisfactorily maintained in this way for more than 8 days. 3. There was no significant difference in the cell sensitivity to RSV between the 15I-line and New York S-line chicken embryo fibroblasts. 4. The cell growth inhibitors in the fresh calf sera employed were destroyed readily by heating the sera at 56℃ for 30 minutes, and diminished during the storage at -25℃ for a certain period but a certain serum was not. 5. Lamb serum was applicable for the cultivation of chicken embryo fibroblasts besides calf serum, but adult cattle and horse serum not. 6. No trouble was encountered against RSV foci forming in the use of calf serum. 7. It was proved that pH 7.3 was to be optimal for the growth of chiken embryo fibroblasts. 8. For dispersing the cultivated cells, the treatmet with 1 x ATV solution was substituted for the trypsinization with 0.05 % trypsin solution (Rubin method). 9. It was better to incubate the primary cultures by the conventional method than by incubation in a CO_2 incubator for the maintenance of monolayer cell sheets. 10. The RSV assay system employed in this study is essentially similar to the original procedure previously reported. However, the virus dose added to the cell cultures, the virus adsorption time applied, the handling after the virus inoculation and the method employed for feeding the inoculated cultures after hard agar overlay are slightly different from the original procedure. This system was connected with the test of embryos for RIF activity, RIF assay and assay of RSV antibody. 11. In the passage of RIF infected cells, it was employed to passage 1 x 10^6 cells under the liquid feed media. By this method the preparation of the plates of 4 x 10^5 cells for the successive challenge of three times with RSV at intervals of 3 days could be successfully made, whereas unsatisfactory performance was made by employing the passage of either 1 x 10^6 or 2 x 10^6 cells under the same media in the preparation of the plates of 1 x 10^6 cells for the successive challenge. 12. In the assay of RSV antibody, the method of inactivation at 4℃ for 18 hours was employed instead of the conventional method (37℃ for 40 minutes) for the contact between the serum to test and the virus, and the serum dilution to be tested was started from the 1:5 dilution of the serum.

Production of Recombinant Human Von Willebrand Factor in the Milk of Transgenic Pigs

LEE, Hyun-Gi,LEE, Hwi-Cheul,KIM, Sung Woo,LEE, Poongyeon,CHUNG, Hak-Jae,LEE, Yun-Keun,HAN, Joo-Hee,HWANG, In-Sul,YOO, Jong-Il,KIM, Yong-Kook,KIM, Hun-Taek,LEE, Hoon-Taek,CHANG, Won-Kyong,PARK, Jin-Ki Society for Reproduction and Development 2009 The Journal of reproduction and development Vol.55 No.5

<P>Von Willebrand factor (vWF), a large multimeric glycoprotein present in blood plasma, is a blood protein of the coagulation system. It is defective in von Willebrand disease and is involved in a large number of other diseases, including thrombotic thrombocytopenic purpura-hemolytic uremic syndrome and heyde's syndrome. We have developed a line of transgenic swine harboring recombinant human von Willebrand factor (rhvWF) cDNA through microinjection of fertilized one-cell pig zygotes. Expression of rhvWF in the mammary gland and secretion of rhvWF into the milk of the transgenic swine were confirmed by immunohistochemical and western blot analyses, respectively, and rhvWF proteins were detected in milk from all lactating founder females at concentrations that were 28- to 56-folds greater than that in circulating human plasma. The amino acid sequence of rhvWF protein in the transgenic pig milk matched that of vWF produced from human blood plasma. This study provides evidence that production of rhvWF from transgenic pig milk is a potentially valuable technology and can be used as a cost-effective alternative in clinical applications.</P>