JaLCDOI 10.18926/AMO/32696
フルテキストURL fulltext.pdf
著者 Okabe, Akinobu| Katayama, Takeshi| Kanemasa, Yasuhiro|
抄録 <p>We examined the method for determining microquantities of lipids, including cholesterol, cholesteryl esters and phospholipids. A standard colorimetric procedure of cholesteryl esters was modified to accommodate a quantitative thin-layer chromatography. This method involved the following steps. (1) Separation of lipids by a thin-layer chromatography: Lipids were applied to Silica gel G plates. Plates were developed with petroleum ether-diethyl etheracetic acid (82: 18: 2, vIvIv). (2) Elution of cholesterol and its esters from scraped silica gel: After scraping the silica gel with adhered cholesterol and its esters, they were eluted with chloroform-methanol (4: 1, v,tv). In the case of phspholipids, the silica gel was calcified. (3) Colorimetric determination of the lipids: Cholesterol and its esters eluted from the silica gel were determined by the method of ZAK with ROSENTHAL'S color reagent directly and after saponification, respectively. Phospholipids were calculated from the phosphorous content determined by the method of KATES. On the basis of examination of recovery and analyses of lipids extracted from tissue, it was concluded that this method permitted a reliable estimation of microquantities of cholesterol, its esters and phospholipids from small amounts of biological materials.</p>
Amo Type Article
発行日 1974-12
出版物タイトル Acta Medica Okayama
28巻
6号
出版者 Okayama University Medical School
開始ページ 403
終了ページ 410
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 4282002
NAID 120002311659
JaLCDOI 10.18926/AMO/32220
フルテキストURL fulltext.pdf
著者 Yoshida, Iwao| Takamizawa, Akihisa| Fujita, Hiyoyuki| Manabe, Sadao| Okabe, Akinobu|
抄録 <p>We constructed a plasmid, pBH103-ME5, in which the region encoding the 10 preS2 amino acid residues and the S domain of the hepatitis B surface antigen (HBsAg) were regulated by the promoter of the yeast repressible acid phosphatase gene. Saccharomyces cerevisiae carrying pBH103-ME5 produced the HBs antigen (yHBsAg), when it was cultured in a medium containing a low concentration of phosphate. The antigen was purified to homogeneity. Its molecular weight was determined by Western blotting to be 24,000, and its amino acid composition agreed well with that deduced from the nucleotide sequence. The C-terminal amino acid sequence of yHBsAg was exactly the same as that predicted from the nucleotide sequence, while the N-terminal amino acid acetylserine, which was followed by 8 amino acid residues coded by the preS2 region. These results indicate that the recombinant yeast produced a single polypeptide consisting of the preS2 region and the subsequent S domain after being processed at the N-terminus</p>
キーワード hepatitis B surface antigen preS2 region plasmid yeast Saccharomyces cerevisiae
Amo Type Article
発行日 1991-02
出版物タイトル Acta Medica Okayama
45巻
1号
出版者 Okayama University Medical School
開始ページ 1
終了ページ 10
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 2063691
Web of Science KeyUT A1991FA75000001
JaLCDOI 10.18926/AMO/32216
フルテキストURL fulltext.pdf
著者 Konobe, Takeo| Ishikawa, Nobuyoshi| Gohda, Hideo| Fukai, Konosuke| Okabe, Akinobu|
抄録 <p>The hepatitis B virus surface antigen containing the preS2 nine amino acid sequence produced by a recombinant Saccharomyces cerevisiae (yHBsAg) was purified and its physicochemical properties were determined. Ultrastructurally, the yHBsAg was found to be a homogeneous spherical particle with a diameter of 24 +/- 4 nm. The homogeneity of the yHBsAg particles was also demonstrated by analyses of their buoyant density and isoelectric point. They consisted of protein (53%), lipid (36%) and carbohydrate (11%), and the alpha-helix content was estimated to be 32%, differing from the reported values for human plasma-derived HBsAg (hHBsAg). Immunodiffusion analysis showed that the antigenic specificity of yHBsAg was identical to that of hHBsAg. Immunization of mice demonstrated that the immunogenicity of the yHBsAg was significantly higher than that of hHBsAg. </p>
キーワード hepatitis B surface antigen yeast Pre S2 immunogenicity recombinant yeast
Amo Type Article
発行日 1991-02
出版物タイトル Acta Medica Okayama
45巻
1号
出版者 Okayama University Medical School
開始ページ 11
終了ページ 19
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 1712147
Web of Science KeyUT A1991FA75000002
JaLCDOI 10.18926/AMO/30903
フルテキストURL fulltext.pdf
著者 Okabe, Akinobu|
抄録 <p>Cholesterol, cholesteryl esters, triglycerides and fatty acids as major neutral lipids and phospholipids were examined in quantitative analysis. The method consisted of three steps: (1) separation of lipids by one-dimensional thin-layer chromatography on silica gel plates; (2) elution of neutral lipids from scraped silica gel with chloroform-methanol (4:1); and (3) colorimetric determination of individual neutral lipids in eluates and phospholipids in silica gel. The conditions were modified for chromotropic acid reaction for determining triglycerides. Laurell's method for determining fatty acids was also modified to apply to quantitative thin-layer chromatography. The accuracy of the modified methods was well-defined as the absorbance values were on a linear curve. A quantitative study was made of the recovery of triglycerides and fatty acids after chromatography. Combining these modified methods and colorimetry for determination of cholesterol cholesteryl esters and phospholipids, the author established a micromethod for determining the major neutral lipids and phospholipids by thin-layer chromatography. Lipids from HeLa, S-3 cells were analyzed to examine the applicability of this method to tissues. The results indicated that the new method permitted a reliable estimation of the major neutral lipids and phospholipids from small amounts of tissues.</p>
Amo Type Article
発行日 1975-08
出版物タイトル Acta Medica Okayama
29巻
4号
出版者 Okayama University Medical School
開始ページ 261
終了ページ 272
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 128990
NAID 120002305475
JaLCDOI 10.18926/AMO/30338
フルテキストURL fulltext.pdf
著者 Minami, Junzabro| Okabe, Akinobu| Nagata, Akihide| Hayashi, Hideo|
抄録 <p>A modified method of passive immune hemolysis (PIH) was applied to the quantitative assay of heat-labile enterotoxin (LT) produced by enterotoxigenic Escherichia coli. The method enabled the measurement of 0.2 to 1.2 ng LT. The production of LT by enterotoxigenic E. coli under various conditions was analyzed using the modified method. LT production was intense during the logarithmic growth phase and decreased during the stationary growth phase. Lincomycin (50 to 100 micrograms/ml) affected cell growth slightly, but enhanced production of LT until the late-stationary growth phase. About 90% of the LT produced was retained in the cell, and the rest was excreted into the culture medium. The initial pH of the culture medium affected LT production. Alkaline pH enhanced LT production, though growth was depressed. Aeration enhanced both growth and LT production.</p>
キーワード heat-labile enterotoxin E coli passive immune hemolysis
Amo Type Article
発行日 1984-10
出版物タイトル Acta Medica Okayama
38巻
5号
出版者 Okayama University Medical School
開始ページ 461
終了ページ 469
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
論文のバージョン publisher
査読 有り
PubMed ID 6393717
Web of Sience KeyUT A1984TQ61100006