JaLCDOI 10.18926/AMO/32643
FullText URL fulltext.pdf
Author Kobayashi, Kazuko| Watarai, Shinobu| Yasuda, Tatsuji|
Abstract <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>
Keywords ganglioside GD3 tumor-associated antigen liposomes antigen determination monoclonal antibody
Amo Type Article
Published Date 1992-12
Publication Title Acta Medica Okayama
Volume volume46
Issue issue6
Publisher Okayama University Medical School
Start Page 435
End Page 441
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 1485537
Web of Science KeyUT A1992KE49600005
JaLCDOI 10.18926/AMO/31966
FullText URL fulltext.pdf
Author Jiang, Yan| Matsuo, Toshihiko| Fujiwara, hirotake| Hasebe, Satoshi| Ohtsuki, Hiroshi| Yasuda, Tatsuji|
Abstract <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>
Keywords congenital superior oblique muscle palsy congenital fibrosis of the extraocular muscles (CFEOM) ARIX PHOX2B polymorphism
Amo Type Article
Published Date 2005-04
Publication Title Acta Medica Okayama
Volume volume59
Issue issue2
Publisher Okayama University Medical School
Start Page 55
End Page 62
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 16049556
Web of Science KeyUT 000228590000004
JaLCDOI 10.18926/AMO/31125
FullText URL fulltext.pdf
Author Zhang, Daxian| Yasuda, Tatsuji| Yu, Yingyan| Okada, Shigeru|
Abstract <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>
Keywords lipid peroxidation liposome metal-chelate complex physicochemical damage
Amo Type Article
Published Date 1994-06
Publication Title Acta Medica Okayama
Volume volume48
Issue issue3
Publisher Okayama University Medical School
Start Page 131
End Page 136
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 7942071
Web of Science KeyUT A1994NV04300003
JaLCDOI 10.18926/AMO/31099
FullText URL fulltext.pdf
Author Umeda, Mamoru| Yasuda, Tatsuji|
Abstract <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>
Keywords liposome immune lysis assay C-reactive protein carboxyfluoescein mouse monoclonal antibodies
Amo Type Article
Published Date 1994-12
Publication Title Acta Medica Okayama
Volume volume48
Issue issue6
Publisher Okayama University Medical School
Start Page 299
End Page 304
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 7535968
Web of Science KeyUT A1994PZ34600003
JaLCDOI 10.18926/AMO/30793
FullText URL fulltext.pdf
Author Zhao, Dan-Dan| Watarai, Shinobu| Lee, Jin-tae| Kouchi, Shuuichi| Ohmori, Hitishi| Yasuda, Tatsuji|
Abstract <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>
Keywords cationic liposome luciferase plasmid DNA transfection efficiency
Amo Type Article
Published Date 1997-06
Publication Title Acta Medica Okayama
Volume volume51
Issue issue3
Publisher Okayama University Medical School
Start Page 149
End Page 154
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 9227794
Web of Science KeyUT A1997XJ12700006
JaLCDOI 10.18926/AMO/30488
FullText URL fulltext.pdf
Author Kobayashi, Kazuko| Han, Mei| Watarai, Shinobu| Yasuda, Tatsuji|
Abstract <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>
Keywords brain targeting liposomes mouse THA(9-amino-1 2 3 4 -tetrahydroacridin)
Amo Type Article
Published Date 1996-04
Publication Title Acta Medica Okayama
Volume volume50
Issue issue2
Publisher Okayama University Medical School
Start Page 67
End Page 72
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 8744931
Web of Science KeyUT A1996UJ08100002
JaLCDOI 10.18926/AMO/30404
FullText URL fulltext.pdf
Author Yonei, Taiji| Watarai, Shinobu| Okada, Yoshio| Yasuda, Tatsuji| Tsuji, Takao|
Abstract <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>
Keywords diabetes nephropathy monoclonal antibody microalbuminuria
Amo Type Article
Published Date 1995-06
Publication Title Acta Medica Okayama
Volume volume49
Issue issue3
Publisher Okayama University Medical School
Start Page 153
End Page 159
ISSN 0386-300X
NCID AA00508441
Content Type Journal Article
language 英語
File Version publisher
Refereed True
PubMed ID 7676846
Web of Science KeyUT A1995RH05400006
Author Yasuda, Tatsuji|
Published Date 1991-08
Publication Title 環境病態研報告
Volume volume62
Content Type Others
JaLCDOI 10.18926/14495
Title Alternative New immunization procedure for production of monoclonal antibodies which recognize carbohydrate of glycoprotein
FullText URL 062_038_045.pdf
Author Ooyama, Kunio| Watarai, Shinobu| Yasuda, Tatsuji|
Abstract 糖タンパク質糖鎖に対するモノクローナル抗体を効率よくとるための免疫方法の検討をおこなった。抗原としては糖鎖のがん性変化のひとつであるbisecting N-acetyiglucosamine構造をもつオボムコイド(OVM)をとりあげた。OVM全分子を通常のフロインド完全アジュバントでくりかえし感作する方法では糖鎖を認識するモノクローナル抗体をとることはできなかった。またOVMをリポソームニ重膜に挿入する方法でも同様であった。これに対してOVMからプロナーゼ消化により糖ペプチドを調製し,アジュバント活性をもつリピドAを共存させたリポソームに共有結合した抗原を感作したマウスからは高率に糖鎖と反応するモノクローナル抗体をとることができた。しかしながらこれらの抗体はいずれもサブタイプはIgMであった。
Abstract Alternative New immunization method for production of monoclonal anitibodies which recognize oligosaccharide portion of glycoprotein was developed. Conventional immunization method in which glycoprotein was emulsified with Freund's complete adjuvant could not produce anti-carbohydrate monoclonal antibody. Glycopeptide which was prepared by pronase digestion of glycoprotein conjugated liposomes containing lipid A of Salmonella minnesota were revealed good antigen for prodution of anti-carbohydrate monoclonal anitibodies. By this new immunization method several monoclonal antibodies which recognize mainly carbohydrate portion of ovomucoide were established.
Keywords モノクローナル抗体 (Monoclonal antibody) 糖タンパク質 (glycoprotein) オボムコイド (ovmucoide) 酵素免疫測定法 (enzyme immunoassay) リポソーム (liposome) lipid A
Publication Title 環境病態研報告
Published Date 1991-08
Volume volume62
Start Page 38
End Page 45
ISSN 0913-3771
language 日本語
File Version publisher
NAID 120002308209
JaLCDOI 10.18926/11648
Title Alternative 種々の長さのスペーサーをもつハプテン化ボスファチジルエタノールアミンの新しい合成法
FullText URL 062_032_037.pdf
Author Ishimori, Yoshio| Yasuda, Tatuji|
Abstract The antigenicity of liposomes sensitized with haptenated phosphatidylethanolamine (PE) and the reactivity of the liposomes with complement depended on the length of the spacer between hapten and PE. To establish the optimal conditions for the assay, haptenated PE's with various length of spacers are required. In the previous method, hapten-spacer molecule was first synthesized to which PE was conjugated. Therefore, even different hapten molecules and different length of spacer molecules were used, every combination of hapten and spacer has to be synthesized. A new procedure for preparing hapten-spacer-PE was described here. We first prepared conjugates between PE and various length of spacer molecule, the terminal of which is an amino residue. These molecules react well with activated hapten molecules, giving a good yield of hapten-spacer-PE.
Abstract Alternative 人工脂質膜であるリボソームにハプテン化ホスファチジルエタノールアミン(PE)を挿入することで,リボソーム膜上での免疫反応の研究が進んでいる。いろいろな因子のなかでリボソーム表面とハプテン基の間のスペーサーの長さも重要な因子であることが判明してきた。このスペーサーの役割を研 究するためには汎用性のある合成法の開発が望まれている。これまでのハプテン基一スペーサー分子を結合する方法は種類の異なるハプテン基をもつ分子群を合成するには煩雑である。そこで種々のスペーサーをもつPEを先に合成することで種類の違うハプテン基をもち,異なるスペーサーをもつハプテン化脂質抗原の合成法を開発した。
Keywords Haptenated phosphatidylethanolamine (ハプテン化ホスファチジルエタノールアミン) Spacer (スペーサー) Liposomes (リポソーム)
Publication Title 環境病態研報告
Published Date 1991-08
Volume volume62
Start Page 32
End Page 37
ISSN 0913-3771
language 英語
File Version publisher
NAID 120002313629