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Biomedical subjects

N Adachi

Publications and source records attributed to N Adachi.

At least 235 records · Page 13Linked to original sources
PubMed1980

New fluorogenic peptide substrates for plasmin.

Fluorogenic peptides, peptidyl-4-methylcoumaryl-7-amides (MCA), containing COOH-terminal lysine residues, were newly synthesized and tested as substrates for plasmin. Among six peptidyl-MCA's, Boc-Val-Leu-Lys-MCA and Boc-Glu-Lys-Lys-MCA were found to be useful for the specific and sensitive assay of plasmin. The Km values estimated from Line-weaver-Burk plots for these substrates using human and bovine plasmins were in the region of 10(-4) M. Boc-Glu-Lys-Lys-MCA was slightly hydrolyzed by bovine plasma kallikrein, and Boc-Val-Leu-Lys-MCA was slightly hydrolyzed by human and hog urinary kallikreins and hog pancreatic kallikrein. However, both of the fluorogenic peptides were essentially unaffected by urokinase, alpha-thrombin, Factor Xa, Factor IXa, Factor XIa, and Factor XIIa. It was confirmed that plasmin hydrolyzed Boc-Glu-Lys-Lys-MCA, cleaving the lysyl-MCA bond, but not the lysyl-lysyl bond. These fluorogenic peptides were resistant to human plasmin activated by streptokinase. Boc-Glu-Lys-Lys-MCA was not hydrolyzed by human plasmin or plasminogen in the presence of more than a 5-fold molar excess of streptokinase. The sensitivity of Boc-Val-Leu-Lys- of more than a 5-fold molar excess of streptokinase. The sensitivity of Boc-Val-Leu-Lys-MCA to human plasmin was also reduced, but plasmin retained 35% of the maximum activity even in the presence of a 20-fold molar excess of streptokinase. These results suggest that streptokinase-plasmin complex has essentially no activity towards Boc-Glu-Lys-Lys-MCA.

Animals↗
PubMed1980

A new fluorogenic substrate method for the estimation of kallikrein in urine.

Kallikrein in human urine was measured using a fluorogenic peptide substrate, proly-phenyl-alanyl-arginine-4-methylcoumaryl-7-amide (Pro-Phe-Arg-MCA). Using 20 microliters of normal urine, kallikrein activity was quantitatively assayed by incubation with a final concentration of 10(-4) M Pro-Phe-Arg-MCA at 37 degrees C for 90 min. The reaction was terminated by adding 50 units of Trasylolr or 20 microliters of 10% acetic acid. Using this method, kallikrein activity was measured in urine from normal subjects and patients with glomerulonephitis and Bartter's syndrome. These values were comparable with the values obtained by radioimmunoassay using bovine low-molecular-weight kininogen. When normal urine was applied to a column of arginine-Sepharose 4B, two activities, Pro-Phe-Arg-MCA hydrolyzing activity and kinin-releasing activity toward bovine low-molecular-weight kininogen, were eluted in the same fraction. These results indicate that the present method is useful for the estimation of kallikrein in urine, in terms of specificity, sensitivity, and simplicity.

Acetates↗
PubMed1980

The binding protein for retinoic acid from rat testis cytosol: isolation and partial characterization.

This study reports the isolation and partial characterization of a soluble protein with binding specificity for retinoic acid from rat testis cytosol. Cytosol, labeled in vitro by incubation with [14C]retinoic acid, was fractionated by a series of procedures that included ion exchange chromatography on DEAE-Sepharose and on DEAE-cellulose, gel filtration on Sephadex (G-50, and preparative polyacrylamide gel electrophoresis. The resulting cytosol bindin; protein for retinoic acid had been purified approximately 16,000-fold, with recovery of 11% of the specifically bound [14C]retinoic acid. The purified binding protein was homogeneous on disc gel electrophoresis at pH 4.5 or in the presence of SDS, but displayed microheterogeneity on isoelectric focusing, where both a major and a minor band with apparent isoelectric points near 4.7 were observed. Purified retinoic acid binding protein had a molecular weight of approximately 14,600, and an ultraviolet absorption spectrum with two maxima near 277 and 346 nm. The fluorescence intensity of retinoic acid bound to the cytosol binding protein was approximately 30-times greater than that of retinoic acid in solution in any of five organic solvents, suggesting that retinoic acid is highly immobilized when bound to the cytosol binding protein. The properties of the rat testis cytosol binding protein for retinoic acid were compared directly with those of the testis cytosol binding protein for retinol and of serum retinol-binding protein. The two cytosol binding proteins were much more similar to each other than to the serum transport protein, from which they differed in a number of properties, including which they differed in a number of properties, including molecular size, affinity for prealbumin, immunological reactivity, and absorption and fluorescence spectral characteristics.

Amino Acids↗
PubMed1978

Vascular pedicle fibular transplantation as treatment for bone tumor.

In 4 cases of bone tumor in which extensive bone defects resulted from removal of the tumorous focus, vascular pedicle free fibular transplantation was performed using microsurgical techniques together with filling the defects with iliac bone. Early bone union was achieved. Atrophy and fracture were avoided. Early postoperative physical therapy was possible. The vessels selected in the recipient site are dependent upon the location of the graft bed, but the anastomosed site of the vessels should be kept outside of the graft bed. It is important to plan the operation so as to avoid motion at the site of callus formation and tension at the anastomosis site.

Adult↗