JaLCDOI 10.18926/AMO/32643
フルテキストURL fulltext.pdf
著者 Kobayashi, Kazuko| Watarai, Shinobu| Yasuda, Tatsuji|
抄録 <p>We developed a sensitive method for detection of glycosphingolipid (GSL) antigen(s) on the cell surface. As a model of GSL antigen, ganglioside GD3 was used. An IgM monoclonal antibody (DSG-1) specific for ganglioside GD3 was preincubated with standard inhibitor liposomes containing ganglioside GD3. Then carboxyfluorescein-entrapped indicator liposomes containing ganglioside GD3 and complement were added. Release of the marker from the indicator liposomes was specifically inhibited by inhibitor liposomes. The assay system was simple, sensitive, reproducible, and semiquantitative. Pg to ng of ganglioside GD3 could be detected. Furthermore, ganglioside GD3 on the cells was investigated with SK-MEL-28 human melanoma cell line and human red blood cells (HRBC). When SK-MEL-28 melanoma with ganglioside GD3 was used as an inhibitor, specific inhibition was observed. However, HRBC without ganglioside GD3 showed no significant inhibition. The marker release was 50% inhibited by 1.4 x 10(6)SK-MEL-28 melanoma cells/ml. The amount of ganglioside GD3/melanoma cell was estimated to be at least 1.1 x 10(-14) g from the standard curve made with the liposomes containing 10% epitope density of ganglioside GD3. This assay system may be useful for detection of GSL antigen on the cell.</p>
キーワード ganglioside GD3 tumor-associated antigen liposomes antigen determination monoclonal antibody
Amo Type Article
発行日 1992-12
出版物タイトル Acta Medica Okayama
46巻
6号
出版者 Okayama University Medical School
開始ページ 435
終了ページ 441
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
OAI-PMH Set 岡山大学
論文のバージョン publisher
査読 有り
Submission Path amo/vol46/iss6/5
PubMed ID 1485537
Web of Sience KeyUT A1992KE49600005
JaLCDOI 10.18926/AMO/31966
フルテキストURL fulltext.pdf
著者 Jiang, Yan| Matsuo, Toshihiko| Fujiwara, hirotake| Hasebe, Satoshi| Ohtsuki, Hiroshi| Yasuda, Tatsuji|
抄録 <p>To identify ARIX gene and PHOX2B gene polymorphisms in patients with congenital superior oblique muscle palsy, 3 exons of the ARIX gene and PHOX2B gene were sequenced by genomic DNA amplification with polymerase chain reaction (PCR) and direct sequencing in 31 patients with congenital superior oblique muscle palsy and in 54 normal individuals. A family with a father and one daughter each having congenital superior oblique muscle palsy was also included in this study. Eleven patients with congenital superior oblique muscle palsy had heterozygous nucleotide changes in the ARIX gene, including 4 patients reported on previously. One patient with atrophy of the superior oblique muscle had a new change of T-4G in the promoter region of the ARIX gene. The other 6 patients had a heterozygous nucleotide change of G153A in the 5'-untranslated region (UTR) of the exon 1 of the ARIX gene. These nucleotide changes of the ARIX gene, taken together, had a significant association with congenital superior oblique muscle palsy(P = 0.0022). One patient and 5 patients had heterozygous nucleotide changes of A1106 C and A1121 C in exon 3 of the PHOX2B gene, respectively, while these changes were absent in the normal individuals. Two patients had both the G153A change in the 5'-UTR of exon 1 of the ARIX gene and the A1121 C change in exon 3 of the PHOX2B gene. In conclusion, the polymorphisms of the ARIX gene and PHOX2B gene may be genetic risk factors for the development of congenital superior oblique muscle palsy.</p>
キーワード congenital superior oblique muscle palsy congenital fibrosis of the extraocular muscles (CFEOM) ARIX PHOX2B polymorphism
Amo Type Article
発行日 2005-04
出版物タイトル Acta Medica Okayama
59巻
2号
出版者 Okayama University Medical School
開始ページ 55
終了ページ 62
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
OAI-PMH Set 岡山大学
論文のバージョン publisher
査読 有り
Submission Path amo/vol59/iss2/4
PubMed ID 16049556
Web of Sience KeyUT 000228590000004
JaLCDOI 10.18926/AMO/31125
フルテキストURL fulltext.pdf
著者 Zhang, Daxian| Yasuda, Tatsuji| Yu, Yingyan| Okada, Shigeru|
抄録 <p>A carboxyfluorescein (CF)-enveloping soybean phosphatidylcholine liposome was used as a model of physicochemical damage of biomembranes. The liposomes were exposed to a metal-chelate complex [2 mM of ferric nitrilotriacetate (FeNTA) or cupric nitrilotriacetate (CuNTA)] plus a reductant (2 mM of ascorbate or various concentrations of reduced glutathione), and CF release from damaged liposomal membranes and the generation of thiobarbituric acid-reactive substances (TBARS) were measured. In the presence of a reducing agent, both FeNTA and CuNTA stimulated markedly CF release and an increase in the TBARS level, while in the absence of a reducing agent both of the chelate complexes showed little CF release and TBARS. The effects of H2O2 addition to the reaction system containing liposome with FeNTA or CuNTA plus ascorbate were also examined. The CF release was slightly increased by the addition of a smaller dose (0.5 mM) of H2O2 and it was inhibited by 8 mM of H2O2. A similar result was obtained in the TBARS test. These results suggest that FeNTA- or CuNTA-mediated lipid peroxidation can damage liposomal membranes physicochemically, and the redox reaction of the chelated metal itself is more important than a Fenton-type reaction in the process.</p>
キーワード lipid peroxidation liposome metal-chelate complex physicochemical damage
Amo Type Article
発行日 1994-06
出版物タイトル Acta Medica Okayama
48巻
3号
出版者 Okayama University Medical School
開始ページ 131
終了ページ 136
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
OAI-PMH Set 岡山大学
論文のバージョン publisher
査読 有り
Submission Path amo/vol48/iss3/3
PubMed ID 7942071
Web of Sience KeyUT A1994NV04300003
JaLCDOI 10.18926/AMO/31099
フルテキストURL fulltext.pdf
著者 Umeda, Mamoru| Yasuda, Tatsuji|
抄録 <p>We have already developed the liposome immune lysis assay (LILA) for the determination of C-reactive protein (CRP) by employing an inhibition method and a sandwich method. We herein report a new LILA system involving the use of monoclonal antibodies-bearing liposomes. We established five monoclonal antibodies to CRP antigen, AC-1, -2, -3, -4, -5 which had the capacity to activate complement and form antigen-antibody complex. Each of these antibodies was covalently coupled to carboxyfluorescein-entrapped multilamellar liposomes. When the liposomes were incubated with CRP antigen in the presence of guinea pig complement, CRP antigen-dependent liposome lysis was observed but the sensitivity was not great enough for practical use. On the other hand, when liposomes coupling two monoclonal antibodies (AC-1, AC-2) which recognized distinct CRP antigenic determinants were employed in the assay, the sensitivity increased compared with that using only one monoclonal antibody, and the detectable concentration range was 5-300 ng/ml. These results indicated that the combination of two or more monoclonal antibodies which recognize distinct CRP antigenic determinants is effective for increasing the sensitivity of the assay.</p>
キーワード liposome immune lysis assay C-reactive protein carboxyfluoescein mouse monoclonal antibodies
Amo Type Article
発行日 1994-12
出版物タイトル Acta Medica Okayama
48巻
6号
出版者 Okayama University Medical School
開始ページ 299
終了ページ 304
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
OAI-PMH Set 岡山大学
論文のバージョン publisher
査読 有り
Submission Path amo/vol48/iss6/3
PubMed ID 7535968
Web of Sience KeyUT A1994PZ34600003
JaLCDOI 10.18926/AMO/30793
フルテキストURL fulltext.pdf
著者 Zhao, Dan-Dan| Watarai, Shinobu| Lee, Jin-tae| Kouchi, Shuuichi| Ohmori, Hitishi| Yasuda, Tatsuji|
抄録 <p>We compared the transfection efficiency of four types of positively charged liposomes composed of (i) N-(&#945;-trimethylammonioacetyl)-didodecyl-D-glutamate chloride (TMAG), dilauroylphosphatidylcholine (DLPC), and dioleoylphosphatidylethanolamine (DOPE) (1:2:2 molar ratio); (ii) 3&#946; [N-(N&#8242;, N&#8242;-dimethylaminoethane)-carbamoyl] cholesterol (DC-Chol) and DOPE (3:2 molar ratio); (iii) dimethyldioctadecylammonium bromide (DDAB) and DOPE (1:2.2 molar ratio); (iv) N-[1-(2,3-dioleyloxy) propyl] -N,N,N-trimethylammonium chloride (DOTMA) and DOPE (1:1, w/w; lipofectin). Luciferase gene was used as a reporter gene. Among the cationic liposomes used, the liposomes composed of TMAG, DOPE and DLPC showed a much higher efficiency of plasmid DNA entrapment than the other cationic liposomes tested. In the absence of serum, the cationic multilamellar vesicles (MLV) and small unilamellar vesicles (SUV) composed of TMAG, DOPE and DLPC gave highly efficient transfection. On the other hand, MLV, dehydration-rehydration vesicles (DRV), and SUV liposomes prepared with the mixtures of DC-Chol and DOPE showed similar levels of transfection efficiency. However, the cationic liposomes composed of DDAB and DOPE showed inferior efficiency, whether in the form of DRV, SUV or MLV. The transfection efficiency of lipofectin was also low. In the presence of serum, on the other hand, a considerable (about 30-50%) amount of transfection activity was still observed at 10% fetal calf serum in the cationic MLV and SUV composed of TMAG, DOPE and DLPC. Cationic MLV, composed of TMAG, DOPE and DLPC, can transfect plasmid DNA, not only in the adherent cell lines but also in the suspension cell lines. These findings indicate that the transfection efficiency of cationic liposomes is affected by the lipid composition, the type of liposome, or the presence or absence of serum. They also indicate that the cationic liposomes containing TMAG, DOPE and DLPC are efficient vectors for gene transfer into cells.</p>
キーワード cationic liposome luciferase plasmid DNA transfection efficiency
Amo Type Article
発行日 1997-06
出版物タイトル Acta Medica Okayama
51巻
3号
出版者 Okayama University Medical School
開始ページ 149
終了ページ 154
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
OAI-PMH Set 岡山大学
論文のバージョン publisher
査読 有り
Submission Path amo/vol51/iss3/6
PubMed ID 9227794
Web of Sience KeyUT A1997XJ12700006
JaLCDOI 10.18926/AMO/30488
フルテキストURL fulltext.pdf
著者 Kobayashi, Kazuko| Han, Mei| Watarai, Shinobu| Yasuda, Tatsuji|
抄録 <p>&#60;P&#62;Phospholipid vesicles, also known as liposomes, were examined for their ability to act as a drug carrier to the brain. 9-Amino-1,2,3,4-tetrahydroacridine (THA), a centrally acting acetylcholinesterase inhibitor, was used as a model drug. THA was encapsulated in dehydration-rehydration vesicles (DRV) composed of egg yolk phosphatidylcholine, cholesterol and dipalmitoyl-phosphatidic acid (molar ratio, 10/10/1) and injected into the heart of mice. The toxicity and side effects of THA were reduced by encapsulation in liposomes. The THA concentration in the mouse brain after injection of THA-encapsulated DRV at a dose of 2 mg/kg remained higher than that of free THA at the same dose. Effective concentration of THA in the brain was also prolonged by the use of liposomes, although accumulation of THA in the spleen and kidney was observed. We, therefore, concluded that liposomes are useful as carriers of drugs to the brain.&#60;/P&#62;</p>
キーワード brain targeting liposomes mouse THA(9-amino-1 2 3 4 -tetrahydroacridin)
Amo Type Article
発行日 1996-04
出版物タイトル Acta Medica Okayama
50巻
2号
出版者 Okayama University Medical School
開始ページ 67
終了ページ 72
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
OAI-PMH Set 岡山大学
論文のバージョン publisher
査読 有り
Submission Path amo/vol50/iss2/3
PubMed ID 8744931
Web of Sience KeyUT A1996UJ08100002
JaLCDOI 10.18926/AMO/30404
フルテキストURL fulltext.pdf
著者 Yonei, Taiji| Watarai, Shinobu| Okada, Yoshio| Yasuda, Tatsuji| Tsuji, Takao|
抄録 <p>Monoclonal antibodies were raised against urine proteins from diabetic patients. An antibody, YO-2, stained three protein bands with apparent molecular weights of 66, 49, and 36 kDa. These bands were not reactive with an anti-human albumin antibody. The urine levels of YO-2-reactive antigen in the normal control were 0.97 +/- 0.37 U/g-Cr (units per gram of urine creatinine) (mean +/- SD). Those of the normo-, micro-, and macroalbuminuric diabetic patients, respectively, were 1.38 +/- 1.36, 2.87 +/- 2.07, and 3.92 +/- 3.33 U/g-Cr. They were significantly higher in the micro- and macroalbuminuric patients. The urine levels of YO-2-reactive antigen had no significant correlation with the urine albumin levels and hemoglobin A1c. We concluded that; a) monoclonal antibody YO-2 recognized a non-albumin urine antigen increasingly excreted in diabetic patients with nephropathy, b) recent glycemic control of diabetes would not significantly affect the urinary excretion rate of YO-2-reactive antigen, and c) the excretion rate and probably the mechanism of YO-2-reactive protein differed from those of albumin. The urine levels of YO-2-reactive antigen could be a clinical marker of diabetic nephropathy.</p>
キーワード diabetes nephropathy monoclonal antibody microalbuminuria
Amo Type Article
発行日 1995-06
出版物タイトル Acta Medica Okayama
49巻
3号
出版者 Okayama University Medical School
開始ページ 153
終了ページ 159
ISSN 0386-300X
NCID AA00508441
資料タイプ 学術雑誌論文
言語 English
OAI-PMH Set 岡山大学
論文のバージョン publisher
査読 有り
Submission Path amo/vol49/iss3/6
PubMed ID 7676846
Web of Sience KeyUT A1995RH05400006
著者 保田 立二|
発行日 1991-08
出版物タイトル 環境病態研報告
62巻
資料タイプ その他
著者 大山 邦夫| 渡来 仁| 保田 立二|
発行日 1991-08
出版物タイトル 環境病態研報告
62巻
資料タイプ 紀要論文
著者 石森 義雄| 保田 立二|
発行日 1991-08
出版物タイトル 環境病態研報告
62巻
資料タイプ 紀要論文