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Biomedical subjects

S Fujii

Publications and source records attributed to S Fujii.

At least 901 records · Page 50Linked to original sources
PubMed1981

High-molecular-weight kininogen from horse plasma. Isolation, characterization and comparison with bovine high-Mr kininogen.

High-molecular-weight (high-Mr) kininogen was purified from horse plasma by chromatography on columns of DEAE-Sephadex A-50, CM-Sephadex C-50, p-chlorobenzylamine-Sepharose and Sephadex G-150. The yield was about 150 mg from 81 of fresh plasma. The purified material gave a single band on sodium dodecylsulfate/polyacrylamide gel electrophoresis and a single precipitin line on immunodiffusion and immunoelectrophoresis. The molecular weight of horse high-Mr kininogen was estimated to be 78000 by dodecylsulfate gel electrophoresis using the Ferguson plot. Its polypeptide content was determined to be 86% by amino acid analysis and there was a total of 581 amino acid residues/molecule of protein. The kininogen contained a total of 13.9% carbohydrates, consisting of hexoses (7.8%), glucosamine (1.9%), galactosamine (0.6%) and sialic acid (3.6%). On incubation of horse high-Mr kininogen with bovine and horse plasma kallikreins, several fragments which contained extremely high levels of histidine, were liberated, in addition to kinin. After the liberation of kinin and histidine-rich fragments, a protein free of kinin and its fragments was isolated. This protein consisted of two polypeptide chains, heavy chain and light chain, which are bridged by disulfide bonds. The molecular weight and amino acid composition of the heavy chain and the light chain from horse high-Mr kininogen were very similar to those of the heavy and light chains from bovine high-Mr kininogen, respectively. From these results, it was revealed that horse high-Mr kininogen is quite similar to bovine high-Mr kininogen in terms of their physicochemical and chemical properties, although they are immunologically distinguishable.

Amino Acids↗
PubMed1981

[Histological studies of multiple subperitoneal nodules produced by prolonged administration of estrogen (author's transl)].

It has been reported that multiple peritoneal fibroids were produced in guinea pigs treated with large doses of estrogens, and the nodules regressed when the steroid stimulant was discontinued. Recently, these experimental nodules were considered to be simulating the disseminated pattern of Leiomyomatosis Peritonealis Disseminata (LPD). Although these experimental nodules were considered to be composed of fibroblasts by light-microscopic studies, LPD was verified to be composed of smooth muscle by electronmicroscopic studies. Therefore, it is indispensable to study these experimental nodules by electronmicroscopy, when we discuss about the similarities between experimental nodules and LPD. Guinea pigs were treated with various doses of estradiol benzoate (E) [40, 80, 100, 200 microgram/day, thrice a week, intramuscular injection]. After treatment for three months, every dose of E produced the lesions simulating the disseminated pattern of LPD: multiple nodules in various sizes, scattered on the surface of the uterine horn, peritoneum, spleen and pancreas, and in the omentum and mesocolon. Ultrastructurally, the nodules were composed of the cells resembling fibroblast. We conjectured that estrogen differentiates and proliferates the cells of the features of fibroblast-like cells from subcoelomic mesenchyme, resulting in the production of multiple superitoneal nodules simulating the disseminated pattern of LPD.

Animals↗
PubMed1981

[Experimental approach to leiomyomatosis peritonealis disseminata--progesterone-induced smooth muscle-like cells in the subperitoneal nodules produced by estrogen (author's transl)].

Our previous report attempted to verify the pathogenesis and histogenesis of leiomyomatosis peritonealis disseminata (LPD) resulted in the production of multiple subperitoneal nodules simulating the disseminated pattern of LPD, when guinea pigs were treated with various doses of estradiol benzoate (E) [40, 80, 100, 200 micrograms/day, thrice a week, intramuscular injection (i.m.)] for a minimum of three months. Ultrastructurally, the nodules were considered to be composed of the cells resembling fibroblast. However, the nodules of clinically observed LPD were considered to contain not only fibroblasts but also smooth muscle and decidua-like cells. Therefore, for the purpose of inducing smooth muscle and decidua-like cells in the previously prepared nodules in the guinea pig, progesterone treatments were added. Guinea pigs pretreated with E (100 micrograms/day, thrice a week, i.m.) for three months followed by the combination treatment E and progesterone (P) [0.5, 1.0 mg/day, once a week, i.m.] for one to three months produced the nodules similar to that produced by estrogen alone. Ultrastructurally, these nodules were considered to be composed of the cells resembling smooth muscle and decidual cell. From these results, I assumed that estrogen differentiates and proliferates the cells of the features of fibroblast-like cells from subcoelomic mesenchyme which have further capacity to differentiate into smooth muscle and endometrial stroma, and they are differentiated into smooth muscle-like cells and decidua-like cells under both influences of estrogen and progesterone, resulting in the production of multiple subperitoneal nodules.

Animals↗
PubMed1980

Purification and properties of rat stomach kallikrein.

Kallikrein (EC 3.4.21.8) was purified from rat stomach by column chromatography on p-aminobenzamidine-Sepharose, DEAE-Sephadex A-50 and Sephadex G-150 and by isoelectric focusing, measuring its activities to hydrolyse L-prolyl-L-phenylalanyl-L-arginine-4-methyl-coumaryl-7-amide and to release kinin from heat-treated rat plasma. the purified stomach kallikrein showed a single band on polyacrylamide gel electrophoresis at pH 7.0. Its molecular weight was calculated to be 29 000 by gel-filtration on a column of Sephadex G-50. The kallikrein was stable between pH 6-11 and hydrolyzed L-prolyl-L-phenylalanyl-L-arginine-4-methyl-coumaryl-7-amide optimally at pH 11.0. The L-prolyl-L-phenylalanyl-L-arginine-4-methyl-coumaryl-7-amide hydrolyzing activity of rat stomach kallikrein was inhibited by diisopropyl fluorophosphate and Trasylol, but not by trypsin inhibitors from soybean, lima bean and ovomucoid. These properties of rat stomach kallikrein are different from those of partially purified rat plasma kallikrein, but similar to those of glandular kallikreins from other species. From these results, it was concluded that kallikrein is present in rat stomach and that it can be classified as a glandular kallikrein.

Animals↗
PubMed1980

A sensitive colorimetric assay for human urinary kallikrein.

L-Prolyl-L-phenylalanyl-L-arginine-alpha-naphthylester (Pro-Phe-Arg-NE) was synthesized as a new substrate for use in the assay of kallikreins. An assay was developed based on the colorimetric determination of alpha-naphthol released by the enzyme reaction. With Pro-Phe-Arg--NE as substrate, the minimum detectable concentration of human urinary kallikrein, was about 0.001 KU. Thus use of Pro-Phe-Arg-NE provides a highly sensitive method for detection of human urinary kallikrein. Kallikrein could be determined with a 25-microliter sample of human urine. Zymograms of human urinary kallikrein were prepared using Pro-Phe-Arg-NE as substrate. Six bands were separated by polyacrylamide disc gel isoelectrophoresis.

Colorimetry↗
PubMed1980

High-pressure liquid chromatographic determination of ftorafur [1-(tetrahydro-2-furanyl)-5-fluorouracil] and GLC-mass spectrometric determination of 5-fluorouracil and uracil in biological materials after oral administration of uracil plus ftorafur.

Oral administration of uracil plus ftorafur [1-(tetrahydro-2-furanyl)-5-fluorouracil] has a greater antitumor effect than that of ftorafur alone. A high-pressure liquid chromatographic methods was developed for the determination of ftorafur in plasma and visceral tissues with a sensitivity of 0.025 microgram/ml or g of wet weight. A combination of GLC--mass fragmentography and GLC--mass spectrometry with total-ion monitoring was developed for the specific, simultaneous determination of 5-fluorouracil, and active metabolite, and uracil as their trimethylsilylated derivatives. The detection limits for 5-fluorouracil and uracil in the first method were 0.001 microgram/ml for plasma and 0.001--0.005 microgram/ml for visceral tissues, and those in the second method were 0.2 microgram/ml or g of wet weight. The precision and sensitivity of the assay appear to be satisfactory for determination of the levels of these compounds in plasma and visceral tissues.

Administration, Oral↗
PubMed1980

Purification and properties of aminopeptidase and arylamidase from human liver.

Aminopeptidase and arylamidase from human liver were purified 1,580- and 1,130-fold, respectively, by ammonium sulfate fractionation, TEAE-cellulose and hydroxylapatite column chromatographies, and Sephadex G-200 gel filtration. The purified preparations appeared to be homogeneous on polyacrylamide gel electrophoresis. The molecular weights of aminopeptidase and arylamidase were determined to be 280,000 and 170,000, respectively, by gel filtration on Sephadex G-200. Aminopeptidase showed marked specificities for amino acid amides, such as L-leucineamide, L-phenylalanineamide, and L-methionineamide. On the other hand, arylamidase showed specificities for amino acid beta-naphylamides, such as L-alanyl-beta-naphthylamide and L-leucyl-beta-naphthylamide, but it also hydrolyzed amino acid amides to a considerable extent. Immunological studies using antibodies to purified aminopeptidase and arylamidase showed that the antigenicities of these two enzymes were different. Neuraminidase treatment of the purified arylamidase changed its isoelectric point from 3.25 to 4.95. Various properties (Km values, optimum pH, substrate specificities, and antigenicity) of the arylamidase were not changed by neuraminidase treatment, but the enzyme became thermo-labile.

Aminopeptidases↗
PubMed1980

A sensitive colorimetric assay for various proteases using naphthyl ester derivatives as substrates.

A convenient and highly sensitive colorimetric assay for various proteases, such as trypsin, chymotrypsin, plasmin, thrombin, and urokinase is described. The substrates used are alpha-naphthyl ester derivatives of N alpha-tosyl-L-lysine, N alpha-acetylglycyl-L-ination of alpha-naphthol released from them. Use of these alpha-naphthyl ester derivatives made the method more sensitive than the use of the corresponding methyl or ethyl ester derivatives. The minimum detectable concentrations of trypsin, chymotrypsin, plasmin, thrombin and urokinase in this method were about 0.002 micrograms, 0.01 microgram, 0.002 CU, 0.01 IU, and 2 IU, respectively. The Km values of trypsin and thrombin for TLNE were 0.11 mM and 0.15 mM while those for TLME were 2.5 mM and 6.7 mM, respectively; the Km values of chymotrypsin for ATNE and ATEE were 0.18 mM and 0.7 mM, respectively; and the Km values of urokinase for AGLNE and AGLME were 0.17 mM and 4 mM, respectively. Zymograms of various proteases were easy to prepare using these alpha-naphthyl ester substrates, and zymograms of trypsin and chymotrypsin were made with TLNE and ATNE, respectively, as substrates.

Chymotrypsin↗
PubMed1980

Acid denaturation steps of Streptomyces subtilisin inhibitor. A proton magnetic resonance study of individual histidine environment.

Resonance positions and intensities of the C(2) protons of histidyl residues (His 43 His 106) were followed in the NMR spectrum of a protein, Streptomyces subtilisin inhibitor (MW 23,000), in the pH range 2-9 at 30 degrees C, in order to clarify the microenvironment of the individual histidyl residues and the acid denaturation processes of the protein. The large difference in the 1H-2H exchange rate between the C(2) protons of the two histidyl residues indicates that, whereas His 106 is well exposed to the solvent, His 43 is highly shielded from the solvent. Protonation of His 106 occurs in the pH range 5-9, giving a pKa of 6.0 +/- 0.1. In contrast, His 43 can not be protonated in the native conformation, but is protonation occurs simultaneously with the acid denaturation of the protein in a narrow pH range 2.7-3.6, with pHmid of transition at 3.25. On the other hand, the denaturation of the His 106 environment starts at pH 5.0 and finishes at pH 2.7 with pHmid of transition at 3.7. The denaturation transitions are reversible with pH for both His 43 and His 106, but the rates of these transitions are slow (less than 25 s-1). The Hill coefficient for the His 106 transition was found to be 0.9 +/- 0.1 in the pH range 3.5-5.0, but to be distinctively larger than unity below pH 3.5. On the other hand, the Hill coefficient for the His 43 transition was found to be 4.4 +/- 0.6 for the whole range of transition (pH 2.7-3.6). These results combined with available information about the crystal structure lead us to conclude that the transition of His 106 in the pH range 3.5-5.0 represents a local denaturation caused by the protonation of one neighboring residue, probably Glu 102, whereas the transition of His 43 occurs as a result of cooperative protonation of several residues probably including Phe 113 and Asp 52, and is a denaturation step accompanied by the destruction of the major hydrophobic core.

Bacterial Proteins↗
PubMed1980

Leiomyomatosis peritonealis disseminata.

The ninth documented case of leiomyomatosis peritonealis disseminata (LPD) combined with pregnancy is presented. Light and electron microscopic studies revealed that the smaller tumors were composed partly of decidual cells and partly of fibroblasts, and the larger tumors contained cells resembling smooth muscle cells. Plasma steroid levels were determined during pregnancy. Estrogen levels were within normal range, but progesterone levels were lower than normal, resulting in relative hyperestrogenism. In the case presented, pregnancy, especially with relative hyperestrogenism, may have been a major cause of LPD.

Adult↗
PubMed1980

Uncontrollable cases of absence.

Forty-three cases of absence seizure were studied from the viewpoint of prognosis. thirty-three had been seizure-free for at least one year until the time of follow-up. The remaining 10 cases were uncontrollable and their plasma levels of the drugs were measured. The following characteristics were more significantly found in the uncontrollable group: (1) late onset (14 years old or later) (2) preceding grand mal seizures (3) accompanying automatisms. The above three factors were found to be interrelated. (4) concomitant grand mal seizures of sleep or diffuse (other than awakening) type (5) psychological disturbances (low intelligence, circuitous character) and (6) chronological shift to the Lennox-Gastaut syndrome.

Adolescent↗
PubMed1980

Optical signals from early embryonic chick heart stained with potential sensitive dyes: evidence for electrical activity.

1. Using an optical method for monitoring membrane potential, spontaneous electrical activity in the very early embryonic chick heart at the 7-9 somite stages was measured. 2. Spontaneous absorption signals from the 7-8 somite embryonic chick hearts stained with a potential sensitive merocyanine-oxazolone dye were demonstrated. The signals were observed also when a merocyanine-rhodanine dye was used. These signals were identified as spontaneous electrical activity in the embryonic heart cells. 3. The action spectrum in absorption of the merocyanine-oxazolone dye was triphasic in early embryonic chick heart with an increase in transmittance from 750 to 700, a decrease from 700 to 600, and an increase from 600 to 525 nm. 4. The magnitude of the signal was about 10(-3) of the resting intensity at 675 nm, with the merocyanine-oxazolone dye. The spontaneous absorption signals had a signal-to-noise ratio of about 10, respectively. 5. The absorption signals were markedly depressed by a higher external K+-concentration, however, were not affected by tetrodotoxin (TTX). 6. The results indicate that spontaneous electrical activity is generated at the 7-9 somite developmental stage before the initiation of heartbeat.

Absorption↗