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      • Growth Experiment of Mycobacterium Leprae in Cultured Mouse Peritoneal Macrophages - 2. In vivo infection and in vitro cultivation of trypsin-purified Myco. Leprae

        용태,유준,Yang, Y.T.,Lew, Joon The Korea Society for Microbiology 1972 大韓微生物學會誌 Vol.7 No.1

        조직배양(組織培養)을 이용(利用)한 마우스복강거식세포내 인나균증식실험(人癩菌增殖實驗)으로서 trypsin-정제인나균(精製人癩菌)을 사용하여 1) 인나균(人癩菌)의 마우스복강내접종(腹腔內接種)에 의(依)하여 야기(惹起)되는 복강거식세포내의 생체내(生體內) 인나균감염(人癩菌感染)에 대(對)한 동역학적양상(動力學的樣相), 2) in vivo infection-in vitro cultivation에 의(依)한 인나균(人癩菌)의 복강거식세포내 증식태도(增殖態度) 그리고 3) 인나균(人癩菌)의 복강내접종(腹腔內接種)으로 인(因)한 마우스비장조직(脾臟組織)의 병리학적변화(病理學的變化)를 구명(究明)코저 본연구(本硏究)를 실시(實施)하였으며 아래의 결론(結論)을 얻었다. 1. 인나균(人癩菌)의 마우스복강내접종후(腹腔內接種後) 복강거식세포배양실시까지의 기간(其間)이 연장(延長)됨에 따라 배양(培養)된 복강거식세포에 있어서 세포질내(細胞質內)에 식균된 항산균수(抗酸菌數)와 항산균(抗酸菌)을 식균한 거식세포수(細胞數)가 각각 계속적(繼續的)으로 현저(顯著)하게 감소(減小)되어감이 관찰(觀察)되었다. 2. 인나균(人癩菌)의 복강내접종후(腹腔內接種後) 5개월(個月)에 이르기까지 생체내(生體內)에 존재(存在)하는 마우스복강거식세포에 있어서의 인나균증식(人癩菌增殖)을 관찰(觀察)할 수 없었다. 3. 인나균(人癩菌)의 복강내접종후(腹腔內接種後) 24시간(時間) 및 1주(週)만에 실시(實施)된 복강거식세포배양을 2 내지(乃至) 3개월간(個月間) 그 배양상태(培養狀態)를 유지(維持)하였던바 배양(培養)된 거식세포(細胞)의 염색표본(染色標本)에서 거식세포내(細胞內) 항산균수(抗酸菌數)의 뚜렷한 증가양상(增加樣狀)을 관찰(觀察)할수 있었다. 4. in vivo infection-in vitro cultivation 수기(手技)로서 배양(培養)된 복강거식세포를 사용(使用)한 총항산균수측정실험(總抗酸菌數測定實驗)에서 조직배양개시(組織培養開始) 9 및 11주(週)에 이르러 배양(培養)된 거식세포내(細胞內) 항산균수증가(抗酸菌數增加)에 대한 양적증거(量的證據)를 얻을수 있었다. 5. 인나균(人癩菌)의 복강내접종(腹腔內接種)으로 야기되는 마우스비장조직(脾臟組織)의 병리학적소견(病理學的所見)은 주로 적수부위(赤髓部位)에 나타난 변성변화(變性變化)이었으며, 균접종후(菌接種後) 5개월(個月)에 이르기까지 마우스비장내(脾臟內)에 있어서의 인나균(人癩菌)의 증식양상(增殖樣狀)을 관찰(觀察)할 수 없었다. To grow Myocbacterium leprae in cultured mouse peritoneal macrophages, studies were made with trypsin-purified Myco. laprae on 1) the dynamics of infection of mouse peritonal macrophages in vivo with Myco. leprae by intraperitoneal inoculation, 2) growth experiment of Myco. leprae in cultured mouse peritoneal macrophages by in vivo infection and in vitro cultivation and 3) the observation of pathological changes in spleens of mice induced by intraperitoneal inoculation of Myco. leprae. Results are summarized as follows; 1. Continuing and significant decreases were observed in the numbers of both acid-fast bacilli in cultured macrophage and of macrophages harboring.acid-fast bacilli by the length of inter vats between the time of intraperitoneal inoculation of Myco. leprae and the time of initiation of macrophage culture. 2. No evidence of multiplication of Myco. leprae in the peritoneal macrophages in vivo was found up to 5 months after intraperitoneal inoculation. 3. With cultures of macrophages made 24 hours and 1 week after intraperitoneal inoculation of Myco. leprae and maintained in vitro up to 2 to 3 months, microscopic examination of the stained preparations of cultured macrophages indicated that an apparent increase in the number of acid-fast bacilli in the macrophages did occur. 4. Quantitative experiment with in vivo infected-in vitro cultured macrophages revealed certain features of increase in the number of total acid-fast bacilli in the cultured macrophages 7 and 9 weeks after initiation of the cultures. 5. Pathological changes in the spleens mice inoculated with Myco. leprae were of mainly degenerative nature in the red pulp. No multiplication of Myco. leprae was observed in the spleens of mice up to 5 months after intraperitoneal inoculation.

      • KCI등재후보
      • Shope Papillona Virus에 의한 가토의 유각종 및 암전환 피부암조직의 Histone 단백비교 연구

        용태,김용숙,최철순,이희성 중앙대학교 의과대학 의과학연구소 1987 中央醫大誌 Vol.12 No.1

        In order to isolate and analyze histone fraction of Shope papolloma virus(SPV)-induced papollomas and papilloma-transformed cancer developed in adult domestic rabbits,a series of experiments were conducted in this study and the results are summarized as follows: 1. Omoculation of SPV into deilated back skin of New Zealand white rabbit resulted in the development of typical papillomas at the site of virus inoculum,SPV, was prepared from papilloma tissue induced in cottontail rabbits by the inculation of SPV and has been kept in 50% buffered glycerin at 4℃ for almost 20 years, and this study firmly established that SPV and its tumor-forming activity is extemely stable under the conditions of the storage. 2. As excepted, spontaneous regression of tumors occured in a high proportion od rabbits carrying SPV-induced papillomas by 8 weeks postinoculation. Of 4 rabbits in which spontaneous regression of papillomas did not occur, there observed no development of cancerous transformation from the papillomas during the proionged period of observation up to 30 weeks following the SPV inoculation. 4. Attempts were unsuccessful to demonstrate the presemce of the tumor-specific antigens and SPV-related antigens in the tissue of "old" papillomas by double agar gel diffusion, counterimmunoelectrophoresis and indirect florescent antibody technic. 5. Electron micrographs of "fresh" and "old" papillomas failed to reveal the presemce of mature SPV virions or virus particles in the tumor tissue examined. 6. Histone fractions, i.e, H1 H2a,H2b,H3and H4, were successfully isolated from "fresh" and "old" papillomas and from normal rabbit skin tissue, and fractions were characterized by amino acid analysis and by polycrylamide disc gel electrophoresis. The major findings of histone studies include the followings: ⅰ) The yields of whole histone were 7.348㎎/g of normal rabbit skin, 14.091㎎/g of "fresh" papilloma and 9.903㎎/g of "old" papilloma tissue, respectively. ⅱ) The yields of DNA were 6.252㎎/g of normal rabbit skin, 16.772㎎/g of "fresh"papilloma and 10.988㎎/g of "old" papilloma tissue, respectively.Consequently, the DNA to histone ratios were 1:1.175 for normal beggit skin, 1:0.840 for "fresh"papilloma and 1:0.901 for "old" papilloma tissue. ⅲ) The relative amiunts of 5 fractions of histrone i.e, H1 H2a,H2b,H3 and H4, were 15.11, 24.97, 32.62, 8.15 and 19.15% for normal rabbit skin, 8.34, 25.26, 41.84,.3.55,and 21.03% for "fresh"papilloma and 9.44,25.03,35.10,4.88and 25.55% for "old" papilloma tissue, respectively.

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