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Biomedical subjects

Y Ohkubo

Publications and source records attributed to Y Ohkubo.

At least 109 records · Page 6Linked to original sources

Characterization of a panel of monoclonal antibodies to human coagulation factor XI and detection of factor XI in Hep G2 cell conditioned medium.

We have produced a panel of ten monoclonal antibodies specific to coagulation factor XI. Western blot analysis demonstrates that 9 of these antibodies react with the heavy chain of factor XI and one with the light chain. Seven of these antibodies inhibit factor XI and factor XIa activity. We have used immobilised monoclonal antibody for the production of factor XI deficient plasma and to purify factor XI to homogeneity with high yield in a simple two-step procedure. These monoclonal antibodies were used to develop highly sensitive immunoassays capable of detecting less than 0.01 mu factor XI antigen ml-1. A strong correlation was found between antigen and activity levels in 11 patients with hereditary deficiency indicating that none was cross-reacting material positive. Cultured Hep G2 cells were found to synthesize small amounts of factor XI antigen and this could also be detected by functional assay and by western blot analysis.

Animals↗

67Ga in transferrin-unbound form is taken up by inflamed liver of mouse treated with CCl4.

In order to investigate whether or not transferrin is involved in the uptake of 67Ga by inflamed liver (acute inflammatory tissues) the uptake of 67Ga by the liver of mice treated with carbon tetrachloride (CCl4) was studied. The serum GPT value reached its maximum on the 1st day after the CCl4 treatment. The uptake of 67Ga by the liver also reached its maximum on the 1st day after the CCl4 treatment and the amount uptaken into inflamed liver was about 6 times that uptaken into normal liver. On the other hand, the uptake of 125I-transferrin into inflamed liver on the 1st day after CCl4 treatment was only about 1.6 times that into normal liver. Moreover, cold Fe3+ decreased the uptake of 67Ga by normal liver but increased the uptake of 67Ga by inflamed liver. These results show that transferrin plays an important role in the uptake of 67Ga by normal liver but not by inflamed liver, i.e. 67Ga in the transferrin-unbound form is preferentially taken up by inflamed liver.

Animals↗

Structural study of cell wall phosphomannan of Candida albicans NIH B-792 (serotype B) strain, with special reference to 1H and 13C NMR analyses of acid-labile oligomannosyl residues.

Chemical structures of manno-oligosaccharides, from biose to heptaose, released from the phosphomannan of Candida albicans NIH B-792 strain (serotype B) by mild acid hydrolysis were investigated. The results of 1H NMR, 13C NMR, and fast atom bombardment mass spectrometry analyses confirmed that these manno-oligosaccharides belong to a homologous beta-1,2-linked series. Although chemical shifts of 1H NMR patterns of these oligosaccharides were considerably too complicated to be assigned, their 13C NMR patterns were sufficiently simple to be interpreted, exhibiting a regular increase of downfield shift of ppm values of the C-1 atom from each mannopyranose residue in proportion to their molecular weights. In order to determine the whole chemical structure of the parent phosphomannan, the acid-stable domain was subjected to acetolysis and then enzymolysis with the Arthrobacter GJM-1 alpha-mannosidase and the resultant manno-oligosaccharides were investigated for their chemical structures by 1H NMR spectroscopy. The results of a precipitin-inhibition test using the beta-1,2-linked manno-oligosaccharides, from biose to hexaose, in comparison with the corresponding isomers containing alpha-1,2 linkage with small amounts of alpha-1,3 linkage, indicated that the haptens possessing the former linkage exhibited much higher inhibitory effects than the corresponding isomers containing the latter linkages did. Based on the present findings, a chemical structure of the phosphomannan of this C. albicans strain was proposed.

Arthrobacter↗

Purification and characterization of alpha-L-fucosidase from Bacillus circulans grown on porcine gastric mucin.

Bacillus circulans isolated from soil was found to produce two types of alpha-L-fucosidase differing in substrate specificity. One was able to liberate L-fucose from porcine gastric mucin (PGM), but not from artificial substrates, including p-nitrophenyl and methyl alpha-L-fucosides, while the other acted not on PGM but on p-nitrophenyl alpha-L-fucoside. The production of the former enzyme was enhanced about 150 times as much by PGM added to the medium as by glucose. The alpha-L-fucosidase acting on PGM was purified from the culture fluid obtained with PGM medium by ammonium sulfate fractionation and subsequent column chromatography. The purified enzyme was found to be homogeneous by PAGE and its molecular weight was estimated to be approximately 285,000. The optimum pH was found to be 5.5 to 6.5 and the stable pH range was 4.5 to 9.0. The enzyme decomposed various blood group O(H) active substances such as PGM, human milk and human saliva, and moreover acted on A-, B-, and O-erythrocytes. The enzyme was shown to cleave alpha-(1----2)-, (1----3)-, and (1----4)-L-fucosidic linkages in various glycoproteins and oligosaccharides, but failed to hydrolyze alpha-(1----6)-L-fucosic linkages in 6-O-alpha-L-fucopyranosyl-N-acetylglucosamine and intact bovine thyroglobulin.

ABO Blood-Group System↗

Studies on analgesic oligopeptides. VI. Further studies of synthesis and biological properties of tripeptide alkylamides, Tyr-D-Arg-Phe-X.

Nine analogs based on a structure of Tyr-D-Arg-Phe-X (X = alkylamides or alkylhydrazide containing electron-withdrawing atoms or groups) were newly synthesized and their biological properties were examined by the opioid receptor binding properties of mu-, delta- and kappa-receptors, guinea-pig ileum (GPI) assay and analgesic activity in the tail pinch test after subcutaneous administration in mice. Analogs with X = NHCF2CF3, Sar-ol, or NH(CH2)2CN showed potent activities in the GPI and analgesic assays and high affinity for mu-receptor. An analog with X = taurinamide was found to possess 4-fold higher mu-receptor selectivity than that of [D-Ala2, MePhe4, Gly-ol5]enkephalin (DAGO). The receptor binding properties of previously reported analogs [Chem. Pharm. Bull., 33, 1528 (1985); ibid., 33, 4865 (1985); ibid., 36, 4834 (1988)] were also examined for overall discussion of the structure-activity relationships of this series of tripeptide amides.

Amino Acid Sequence↗

Solid phase synthesis and opioid receptor binding activities of [D-Ala2, D-Leu5]enkephalin analogs containing a fluorinated aromatic amino acid.

Five [D-Ala2, D-Leu55]enkephalin (DADLE) analogs containing fluorinated Tyr1 or Phe4 residue, that is, [Phe(2F)4] (I), [Phe(3F)4] (II), [Phe(4F)4] (III), [Tyr(3F)1] (IV) and [Tyr(2F)1] (V), were synthesized by the solid phase method and their opioid receptor affinities were examined. Affinity profiles of five derivatives for the mu- and delta-receptor were similar to those of DADLE, and the affinity for kappa-receptor was zero or negligible.

Enkephalin, Leucine-2-Alanine↗

Inhibition by sodium citrate on 67Ga adsorption to various materials.

The adsorption of 67Ga to the inner surface of stainless steel, polyethylene and silicone tubings was studied. The three tubings remarkably adsorbed 67Ga in order of stainless steel, silicone and polyethylene tubing. Sodium citrate inhibited the adsorption of 67Ga in a dose-dependent manner. These phenomena were also observed in blood sampling via the catheter inserted into the external jugular vein. These results show that constant concentration of sodium citrate is necessary for the in vitro study of 67Ga in order to prevent the adsorption to the surface of experimental materials.

Adsorption↗

The influences of dexamethasone and indomethacin on 67Ga uptake by normal tissues.

Dexamethasone (DEX) increased 67Ga uptake by the liver and spleen at 4, 8, and even 24 h after the injection of 67Ga. These results showed that DEX influenced 67Ga accumulation as well as the initial entry of 67Ga in the liver and spleen. On the other hand, indomethacin (IM) decreased 67Ga uptake by the liver and spleen at 4 h after the injection of 67Ga but did not influence the uptake at 8 or 24 h after the injection of 67Ga. Moreover, DEX or IM little influenced 67Ga uptake by the kidney and muscle. These results suggest that the influence of DEX or IM on 67Ga uptake or accumulation is specific for the liver and spleen.

Animals↗

[MR imaging of splenic masses].

It has been reported that MR imaging of the spleen is unsuccessful in detecting focal lesions because there is not a significant difference in relaxation times between most tumors and surrounding normal spleen. We reviewed the MR imaging of 15 patients (5 cysts, 2 abscesses, 1 hemangioma, 5 malignant lymphomas, 2 metastatic tumors). In all cases, the difference in signal intensities between splenic tissue and mass lesions permitted detection of splenic lesions on MR images. But, malignant lesions were less visible than benign lesions.

Adult↗

[CT findings of the benign tracheobronchial lesions with calcification].

In the benign tracheobronchial lesions with calcification, tracheobronchopathia osteochondroplastica, relapsing polychondritis and tracheobronchial amyloidosis were considered. CT demonstrated small nodules with calcifications at the trachea with or without deformity of tracheal wall in the case of tracheobronchopathia osteochondroplastica, swelling of tracheal cartilage with diffuse and multiple calcifications in the case of relapsing polychondritis and calcifications in the deep parts of tracheobronchial amyloid nodules. CT findings were able to differentiate those benign lesions. High-resolution CT is more useful in the distribution of abnormal calcification of these diseases.

Aged↗

Structural study of phosphomannan of yeast-form cells of Candida albicans J-1012 strain with special reference to application of mild acetolysis.

Structural analysis of the phosphomannan isolated from yeast-form cells of a pathogenic yeast, Candida albicans J-1012 strain, was conducted. Treatment of this phosphomannan (Fr. J) with 10 mM HCl at 100 degrees C for 60 min gave a mixture of beta-1,2-linked manno-oligosaccharides, from tetraose to biose plus mannose, and an acid-stable mannan moiety (Fr. J-a), which was then acetolyzed by means of an acetolysis medium, 100:100:1 (v/v) mixture of (CH3CO)2O, CH3COOH, and H2SO4, at 40 degrees C for 36 h in order to avoid cleavage of the beta-1,2 linkage. The resultant manno-oligosaccharide mixture was fractionated on a column of Bio-Gel P-2 to yield insufficiently resolved manno-oligosaccharide fractions higher than pentaose and lower manno-oligosaccharides ranging from tetraose to biose plus mannose. The higher manno-oligosaccharide fraction was then digested with the Arthrobacter GJM-1 alpha-mannosidase in order to cleave the enzyme-susceptible alpha-1,2 and alpha-1,3 linkages, leaving manno-oligosaccharides containing the beta-1,2 linkage at their nonreducing terminal sites, Manp beta 1----2Manp alpha 1----2Manp alpha 1----2Manp alpha 1----2Man, Manp beta 1----2Manp beta 1----2Manp alpha 1----2Manp alpha 1---- 2Manp alpha 1----2Man, and Manp beta 1----2Manp beta 1----2Manp beta 1----2Manp alpha 1---- 2Manp alpha 1----2Manp alpha 1----2Man. However, the result of acetolysis of Fr. J-a by means of a 10:10:1 (v/v) mixture of (CH3CO)2O, CH3COOH, and H2SO4 at 40 degrees C for 13 h was significantly different from that obtained by the mild acetolysis method; i.e., the amount of mannose was apparently larger than that formed by the mild acetolysis method. In summary, a chemical structure for Fr. J as a highly branched mannan containing 14 different branching moieties was proposed.

Acetic Anhydrides↗

Structural analysis of phospho-D-mannan-protein complexes isolated from yeast and mold form cells of Candida albicans NIH A-207 serotype A strain.

The immunochemical properties between phospho-D-mannan-protein complexes of yeast (Y) and mycelial (M) forms of Candida albicans NIH A-207 (serotype A) strain were compared. Hydrolysis of the Y-form complex gave a mixture of beta-(1----2)-linked D-mannooligosaccharides consisting mainly of tri- and tetra-ose, whereas the M-form complex gave preponderantly D-mannose. The antiserum against Y-form cells exhibited a lower reactivity with the M-form than with the Y-form complex, whereas the antiserum to M-form cells could not distinguish significantly between both complexes. Moreover, these acid-modified complexes showed lower antibody-precipitating effect than each corresponding intact complex against antisera of Y- and M-form cells. Digestion of the acid-modified Y- and M-form complexes with the Arthrobacter GJM-1 strain alpha-D-mannosidase yielded 35- and 40-% degradation products, respectively. Acetolysis of each modified complex under mild conditions gave the same D-mannohexaose, beta-D-Manp-(1----2)-beta-D-Manp-(1----2)-alpha-D-Manp -(1----2)-alpha-D-Manp- (1----2)-alpha-D-Manp-(1----2)-D-Man. Because the complexes of Y- and M-form cells of C. albicans NIH B-792 (serotype B) strain did not give any hexaose fraction containing beta-(1----2) linkages, the presence of this hexaose can be regarded as one of the dominant characteristics of the serotype-A specificity of C. albicans spp.

Candida albicans↗

111In (III) uptake by inflammatory and normal tissues.

Tissue distributions of 111In (III) in the rats bearing granuloma, inflammatory tissue induced by turpentine oil, were compared with those of 67Ga. The results showed that indium-111 resembles 67Ga in the manner of uptake by inflammatory and normal soft tissues. The effect of cold-InCl3 on 111In (III) uptake showed that transferrin is not involved in the uptake of 111In (III) into inflammatory tissues but is involved in the uptake into liver and spleen.

Animals↗

Involvement of transferrin in the uptake of 67Ga in inflammatory and normal tissues.

The results from gel chromatography and electrophoresis showed that 67Ga is exclusively bound with transferrin both in vitro and in vivo, but high concentrations of sodium citrate strongly inhibited the binding of 67Ga to transferrin. The influence of sodium citrate on the uptake of 67Ga into inflammatory and normal soft tissues was also investigated. Sodium citrate decreased the uptake of 67Ga into the liver and spleen, but had no influence on the uptake of 67Ga into inflammatory tissue. These results suggest that the uptake of 67Ga into normal soft tissues occurs in a transferrin-bound form but into inflammatory tissue in an unbound form.

Animals↗

Effect of gallium nitrate on the in vivo distribution of 67Ga in rats bearing inflammatory lesions.

Effect of gallium nitrate on the pharmacokinetics and tissue accumulation of 67Ga was investigated in rats bearing turpentine oil-induced abscess. Gallium nitrate accelerated the blood disappearance of 67Ga, but this effect was less potent than that of ferric nitrate. The accumulation of 67Ga in liver was significantly lowered by gallium nitrate, but no significant decrease of 67Ga accumulation in abscess was observed. On the other hand, 67Ga accumulation in bone was slightly facilitated by gallium nitrate. Ferric nitrate exerted a similar effect on tissue accumulation of 67Ga as gallium nitrate.

Abscess↗