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K Sugahara

Publications and source records attributed to K Sugahara.

At least 19 recordsLinked to original sources
PubMed1992

Determination of cystathionine and perhydro-1,4-thiazepine-3,5-dicarboxylic acid in the urine of a patient with cystathioninuria using column liquid chromatography-mass spectrometry.

A method for the measurement of cystathionine and perhydro-1,4-thiazepine-3,5-dicarboxylic acid in the urine of a patient with cystathioninuria has been developed, using column liquid chromatography-mass spectrometry. Cystathionine and perhydro-1,4-thiazepine-3,5-dicarboxylic acid were determined by scanning the [M + H]+ ions of each compound. The recoveries were 80-92.4% for cystathionine and 80-100% for perhydro-1,4-thiazepine-3,5-dicarboxylic acid after ion-exchange treatment. The results agreed well with those obtained using an amino acid analyser. The concentrations found for cystathionine and perhydro-1,4-thiazepine-3,5-dicarboxylic acid were 1.289 +/- 0.099 mg/ml and 0.310 +/- 0.0067 mg/ml, respectively.

Cystathionine↗
PubMed1992

Identification of the reaction products of the purified hyaluronidase from stonefish (Synanceja horrida) venom.

Recently we purified to homogeneity hyaluronidase from stonefish (Synanceja horrida) venom, for the first time from a marine source [Poh, Yuen, Chung & Khoo (1992) Comp. Biochem. Physiol., in the press]. In the present study the reaction products of the hyaluronidase purified from stonefish venom were analysed. It produced tetra-, hexa-, octa- and deca-saccharides as major end products, but not disaccharides. The structure of the tetrasaccharide product was determined by enzymic analysis, in conjunction with h.p.l.c. and by 1H n.m.r., as GlcA beta 1-3GlcNAc beta 1-4GlcA beta 1-3GlcNAc. Chemical shifts of the structural-reporter-group protons of the constituent monosaccharides for the tetrasaccharide have been assigned. The enzyme did not act on chondroitin sulphate or dermatan sulphate. The results indicate that the stonefish hyaluronidase is an endo-beta-N-acetylglucosaminidase specific for hyaluronate.

Animals↗
PubMed1992

Structural studies on sulfated oligosaccharides derived from the carbohydrate-protein linkage region of chondroitin 6-sulfate proteoglycans of shark cartilage. I. Six compounds containing 0 or 1 sulfate and/or phosphate residues.

Shark cartilage proteoglycans bear predominantly chondroitin 6-sulfate. After exhaustive protease digestion, reductive beta-elimination, and subsequent chondroitinase ABC digestion, 13 hexasaccharide alditols, which are nonsulfated, sulfated, and/or phosphorylated, were obtained from the carbohydrate-protein linkage region. Six compounds, containing 0 or 1 sulfate and/or phosphate residue, represent approximately 40% of the isolated linkage hexasaccharide alditols. They were analyzed by chondroitinase ACII or alkaline phosphatase digestion in conjunction with high performance liquid chromatography, and by 500 MHz one- and two-dimensional 1H NMR spectroscopy. All six compounds have the conventional structure in common. Delta 4,5-GlcA beta 1-3GalNAc beta 1-4GlcA beta 1-3Gal beta 1-3Gal beta 1-4Xyl-ol One compound has no sulfate nor phosphate. Two of the monosulfated compounds have a O-sulfate on C-6 or on C-4 of the GalNAc residue. The third monosulfated compound has a novel O-sulfate on C-6 of the Gal residue attached to xylitol. The two phosphorylated compounds have O-phosphate on C-2 of Xyl-ol, and one of them has in addition sulfate on C-6 of GalNAc.

Animals↗
PubMed1992

Structural studies on sulfated oligosaccharides derived from the carbohydrate-protein linkage region of chondroitin 6-sulfate proteoglycans of shark cartilage. II. Seven compounds containing 2 or 3 sulfate residues.

Shark cartilage proteoglycans bear predominantly chondroitin 6-sulfate. After exhaustive protease digestion, reductive beta-elimination and subsequent chondroitinase ABC digestion, 13 hexasaccharide alditols were obtained from the carbohydrate-protein linkage region and six of them contain 0 or 1 sulfate and/or 1 phosphate residue (Sugahara, K., Ohi, Y., Harada, T., de Waard, P., and Vliegenthart, J. F. G. (1992) J. Biol. Chem. 267, 6027-6035). The other seven compounds, which represent approximately 60% of the isolated linkage hexasaccharides, were analyzed by chondroitinase ACII digestion in conjunction with high performance liquid chromatography and by 500-MHz one- and two dimensional 1H NMR spectroscopy. All seven compounds have the following conventional structure in common. [formula: see text] Two disulfated compounds have an O-sulfate on C-6 of the Gal-2 residue attached to xylitol in combination with an O-sulfate on C-4 or on C-6 of the GalNAc residue. The third disulfated compound has O-sulfate on C-6 of Gal-2, and also on C-6 of Gal-3. Two of the trisulfated compounds also have O-sulfate on C-6 of both Gal-2 and Gal-3 with in addition sulfate on C-6 or C-4 of GalNAc. The other two trisulfated compounds have O-sulfate on C-6 of Gal-2 and on C-4 of Gal-3 in conjunction with sulfate on C-6 or C-4 of GalNAc.

Animals↗
PubMed1992

The phosphorylated and/or sulfated structure of the carbohydrate-protein-linkage region isolated from chondroitin sulfate in the hybrid proteoglycans of Engelbreth-Holm-Swarm mouse tumor.

The structure of the linkage region of chondroitin sulfate chains attached to the hybrid proteoglycans of the Engelbreth-Holm-Swarm mouse tumor was investigated. The peptidoglycan fraction which contains oversulfated chondroitin sulfate rich in the GlcA beta 1-3GalNAc-4,6-diO-sulfate unit and undersulfated heparan sulfate rich in GlcA beta 1-4GlcNAc and GlcA beta 1-4GlcN-2N-sulfate units was isolated after exhaustive protease digestion of the acetone powder of the tumor tissue, (GlcA, glucuronic acid; GalNAc, 2-deoxy-2-N-acetylamino-D-galactose). Glycosaminoglycans were released by beta-elimination using NaB3H4 and digested with chondroitinase ABC. The linkage region fraction was separated from heparan sulfate by gel filtration and fractionated by HPLC on an amine-bound silica column. Six radiolabeled compounds (L1-L6) were obtained and structurally analyzed by cochromatography with authentic hexasaccharide alditols recently isolated by us from the linkage region, and by digestion using chondroitinase ACII, alkaline phosphatase and beta-galactosidase in conjugation with HPLC. These compounds shared the conventional hexasaccharide backbone structure: delta GlcA beta 1-3GalNAc beta 1-4GlcA beta 1-3Gal beta 1-3Gal beta 1-4Xyl-ol, (delta GlcA, delta 4.5-GlcA or D-gluco-4-enepyranosyluronic acid). L1 was not sulfated or phosphorylated. L2 and L4 were monosulfated at C-6 and C-4 of the GalNAc residue, respectively. Upon alkaline phosphatase digestion, L3, L5 and L6 were converted to L1, L2 and L4, respectively. Analysis of the periodate oxidation products indicated that the phosphate group in L3, L5 and L6 is located at C-2 of Xyl-ol. These results suggest that Xyl-2-O-phosphate is associated with both 4-O-sulfated and 6-O-sulfated GalNAc units and does not directly determine the sulfation pattern of chondroitin sulfate.

Animals↗
PubMed1992

A novel sulfated structure in the carbohydrate-protein linkage region isolated from porcine intestinal heparin.

A preparation of porcine stage 14 intestinal heparin, which contains Ser as a predominant amino acid, was used for isolation of the carbohydrate-protein linkage region of heparin. Two glycoserines were isolated in a molar ratio of 96:4 after an exhaustive digestion with a mixture of bacterial heparinase and heparitinases. Their structures were determined by composition analysis, heparitinase digestion, co-chromatography with an authentic glycoserine on high performance liquid chromatography, and by 500-MHz one- and two-dimensional 1H NMR spectroscopy. The structure of the major one is delta GlcA beta 1-3Gal beta 1-3Gal beta 1-4Xyl beta 1-O-Ser and that of the minor is delta GlcA beta 1-4GlcNAc(6-O-sulfate) alpha 1-4GlcA beta 1-3Gal beta 1-3Gal beta 1-4Xyl beta 1-O-Ser. The novel 6-O-sulfated GlcNAc residue was demonstrated to occur in the vicinity of the carbohydrate-protein linkage region. The Gal residues were nonsulfated, in contrast to the sulfated Gal structures recently discovered in the carbohydrate-protein linkage region of chondroitin sulfate proteoglycans. The structural features are discussed in relation to biosynthetic mechanisms of the heparin glycosaminoglycans.

Animals↗
PubMed1992

Surgical treatment of primary carcinoma of the gallbladder based on the histologic analysis of 48 surgical specimens.

During the past 5 years, extended cholecystectomy and/or more radical procedures, including pancreatoduodenectomy and extended hepatic lobectomy, were performed on 48 patients with carcinoma of the gallbladder in an attempt to cure the disease. It was retrospectively proven from microscopic analysis of the surgical specimens taken from the 48 patients that 28 patients had received curative resections. The 28 consisted of all 4 patients with stage I carcinoma, all 9 with stage II, 6 of 8 with stage III, and 9 of 27 with stage IV. We reached the following conclusions: (1) extended cholecystectomy with resection of the bile duct in the hepatoduodenal ligament is the optimum procedure for patients with stage I and stage II carcinomas, and (2) for patients with stage III carcinoma, more radical procedures, including pancreatoduodenectomy and/or wider resection of the liver, might result in curative resection of the disease. For some patients with stage IV tumors, such as duodenal invasion or direct invasion of the liver, curative resection may be achieved by pancreatoduodenectomy or extended hepatic lobectomy.

Adult↗
PubMed1992

Involvement of food intake in the decreased energy retention associated with single deficiencies of lysine and sulphur-containing amino acids in growing chicks.

1. Growth and energy utilisation were determined in growing chicks fed ad libitum on diets deficient either in lysine (5.95 g/kg) or in sulphur-containing amino acids (SAA, 3.5 g/kg). Food intake, body weight gain, energy retained as protein and as fat, and total energy retention were significantly (P less than 0.05) reduced by single deficiencies of either lysine or SAA. 2. Another two experiments were conducted to determine if the decreased total energy retentions in chicks fed on diets deficient in lysine (experiment 3) or SAA (experiment 4) were associated with reduced food intake, by using tube-feeding to control the amount and pattern of food consumption. Chicks fed on diets deficient in lysine or SAA retained less energy as protein and more energy as fat than the control chicks. Neither total energy retention nor heat increment was affected by these deficiencies. Total energy retention was proportional to metabolisable energy (ME) intake alone. 3. It is concluded that the decreased total energy retentions caused by single deficiencies of lysine and SAA were associated with decreased food intake.

Amino Acids, Sulfur↗
PubMed1992

One- and two-dimensional 1H-NMR characterization of two series of sulfated disaccharides prepared from chondroitin sulfate and heparan sulfate/heparin by bacterial eliminase digestion.

The 1H-NMR spectra of eight unsaturated disaccharides obtained by bacterial eliminase digestion of chondroitin sulfate and of heparan sulfate/heparin were recorded in order to construct an NMR data base of sulfated oligosaccharides and to investigate the effects of sulfation on the proton chemical shifts. These shifts were assigned by two-dimensional HOHAHA (homonuclear Hartmann-Hahn) and COSY (correlation spectroscopy) methods. The results indicated the following. (1) Two sets of proton signals were observed, corresponding to the alpha and beta anomers of these disaccharides, except those containing N-sulfated GlcN (2-deoxy-2-amino-D-glucose), in which only one set of signals appeared, corresponding to the alpha anomer. (2) Signals of protons bound to an O-sulfated carbon atom and those bound to the immediately neighboring carbon atoms were shifted downfield by 0.4-0.7 and 0.07-0.3 ppm, respectively. (3) For the disaccharides containing the N-sulfated GlcN, the signals of the protons bound to C-2 and C-3 were shifted upfield by 0.6 and 0.15 ppm, respectively, but that of C-1 was shifted downfield by 0.25 ppm when compared with those of the corresponding N-acetylated disaccharides. (4) For the chondroitin sulfate disaccharides sulfated on the C-4 position of GalNAc (2-deoxy-2-N-acetylamino-D-galactose) or the C-2 position of delta GlcA (D-gluco-4-ene-pyranosyluronic acid), the signal of the H-3 proton of delta GlcA or the H-4 proton of GalNAc was shifted upfield by 0.1-0.15 ppm, indicating the steric interaction of the two sugar components. (5) These effects of sulfation on chemical shifts are additive.

Animals↗
PubMed1992

Overexpression of pulmonary surfactant apoprotein A mRNA in alveolar type II cells and nonciliated bronchiolar (Clara) epithelial cells in streptozotocin-induced diabetic rats demonstrated by in situ hybridization.

Pulmonary surfactant is critical for gas exchange and is composed of both phospholipids and specific surfactant-associated proteins. The most abundant surfactant protein is termed surfactant apoprotein A (SP-A). This protein is thought to be important in the formation of tubular myelin, in absorption of surfactant to the air-liquid interface, in recycling of surfactant in alveolar type II cells, and in the regulation of secretion. We have examined the expression and localization of SP-A mRNA in streptozotocin-induced diabetic rats by in situ hybridization using a specific rat cDNA probe. Diabetes was induced by intraperitoneal injection of 60 mg/kg streptozotocin. After 10 wk, lungs were excised and examined by in situ hybridization and by light and electron microscopy. The ultrastructural examination demonstrated the marked changes of endoplasmic reticulum of alveolar type II cells, as reported previously. Immunohistostaining of SP-A in diabetic lungs was weak in alveolar type II cells. However, by autoradiographs of in situ hybridization, compared with the control lungs, a larger number of silver grains for the SP-A mRNA were shown in alveolar type II cells and also in some bronchiolar epithelial (Clara) cells from the diabetic lungs. Alveolar type II cells having high contents of silver grains were also increased in number. These results were confirmed by measurement of the SP-A content and by Northern blot analysis. The present study demonstrates an overexpression of SP-A mRNA despite the ultrastructural changes in the endoplasmic reticulum of alveolar type II cells in the diabetic lungs, which will provide new information on the regulatory mechanism of SP-A gene expression.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Structural studies on sulfated oligosaccharides derived from the carbohydrate-protein linkage region of chondroitin sulfate proteoglycans of whale cartilage.

From the carbohydrate-protein linkage region of whale cartilage proteoglycans, which bear predominantly chondroitin 4-sulfate, one nonsulfated, two monosulfated and one disulfated hexasaccharide alditols were isolated after exhaustive digestions with Actinase E and chondroitinase ABC, and subsequent beta-elimination. Their structures were analyzed by chondroitinase ACII digestion in conjunction with HPLC and by 500-MHz 1H-NMR spectroscopy. The nonsulfated compound (A) had the following conventional structure: delta GlcA(beta 1-3)-GalNAc(beta 1-4)GlcA(beta 1-3)Gal(beta 1-4)Xylol, where GlcA, delta GlcA and GalNAc are glucuronic acid; 4,5-unsaturated glucuronic acid and 2-deoxy-2-N-acetylamino-D-galactose, respectively. The other compounds were sulfated derivatives of compound A. Two monosulfated compounds (B and C) had an ester sulfate on C4 or C6 of the GalNAc residue, respectively and the disulfated compound (D) had two ester sulfate groups, namely, one on C4 of the GalNAc and the other on C4 of the Gal residue substituted by GlcA. The molar ratio of A/B/C/D was 0.21:0.16:0.36:0.27. The compound containing Gal-4-O-sulfate was previously isolated by us in the form of a sulfated glycoserine [delta GlcA(beta 1-3)GalNAc(4-O- sulfate)(beta 1-4)GlcA(beta 1-3)Gal(4-O-sulfate)(beta 1-3)-Gal(beta 1- 4)Xyl beta 1-O-Ser] from the carbohydrate-protein linkage region of rat chondrosarcoma chondroitin-4-sulfate proteoglycans [Sugahara K., Yamashina, I., DeWaard, P., Van Halbeek, H. & Vliegenthart, J.F.G. (1988) J. Biol. Chem. 263, 10,168-10,174]. The discovery of this structure in the carbohydrate-protein linkage region of chondroitin 4-sulfate proteoglycans from nontumorous cartilage indicates that it is not a tumor-associated product but rather a physiological biosynthetic product since it represents a significant proportion. The biological significance of this structure is discussed in relation to glycosaminoglycan biosynthesis.

Animals↗
PubMed1991

Liquid chromatographic-mass spectrometric analysis for screening of patients with cystinuria, and identification of cystine stone.

Analyses of amino acids in the urine of a normal human and of patients with heterozygous and homozygous cystinuria have been carried out, using liquid chromatography-mass spectrometry with an atmospheric pressure ionization interface system. A kidney cystine stone was also analysed by this system. Very intense quasi-molecular ions ([M + H]+) of standard cystine, arginine, lysine and ornithine were observed on mass chromatograms as base peaks. Mass chromatograms of the urine samples from a normal human and from patients with heterozygous and homozygous cystinuria were easily distinguishable. The retention times in the mass chromatogram and mass spectrum of kidney stone cystine was almost the same as that of authentic cystine.

Arginine↗
PubMed1991

Liquid chromatography-mass spectrometry for simultaneous analyses of iminodipeptides containing an N-terminal or a C-terminal proline.

Simultaneous analyses of synthetic iminodipeptides containing an N-terminal proline or a C-terminal proline have been demonstrated using liquid chromatography-mass spectrometry with an atmospheric pressure ionization interface system. The separation of iminodipeptides was carried out on a reversed-phase high-performance liquid chromatographic column using 0.1% aqueous trifluoroacetic acid-methanol (75:25, v/v, pH 2.0) as mobile phase. Very intense protonated molecular ions [M + H]+ of various synthetic iminodipeptides, Pro-Gly, Gly-Pro, Pro-Ala, Ala-Pro, Pro-Val, Val-Pro, Pro-Leu and Leu-Pro, were observed. Pro-Gly (Pro-X) and Gly-Pro (X-Pro) have the same protonated molecular ion (m/z 173), but the peaks of these compounds on the mass chromatograms were clearly distinguished by the differences of the retention times and mass spectra. The synthetic iminodipeptides containing an N-terminal proline added to urine samples from a patient with prolidase deficiency were also distinguished from iminodipeptides containing a C-terminal proline in urine samples from a patient with prolidase deficiency by scanning the [M + H]+ ion of each iminodipeptide. We established the method to measure simultaneously the various iminodipeptides containing an N-terminal or a C-terminal proline in biological samples.

Amino Acid Sequence↗
PubMed1991

Identification of NAc-HCPC and NAc-beta-CEC, and qualitative analyses of sulphur amino acids in the urine of a patient with cystathioninuria using liquid chromatography/atmospheric pressure ionization mass spectrometry.

Standard sulphur amino acids and various cystathionine metabolites in the urine of a patient with cystathioninuria were analysed using liquid chromatography/mass spectrometry with an atmospheric pressure ionization interface system. Very intense quasi-molecular ions ([M + H]+) of synthetic cystathionine, N-monoacetylcystathionine, perhydro-1,4-thiazepine-3,5-dicarboxylic acid, S-(3-hydroxy-3-carboxy-n-propyl)cysteine, S-(2-carboxyethyl) cysteine, S-(2-hydroxy-2-carboxyethyl)homocysteine, S-(carboxymethyl)homocysteine, N-acetyl-S-(3-hydroxy-3-carboxy-n-propyl)cysteine and N-acetyl-S-(2-carboxyethyl)cysteine were observed by this method. Quasi-molecular ions ([M + H]+) of these sulphur amino acids were observed in the urine sample of the patient with cystathioninuria, and N-acetyl-HCPC and N-acetyl-beta-CEC as N-substituted sulphur amino acids were also identified in the urine of the same patient.

Acetylcysteine↗
PubMed1991

Lung water accumulation in rats after repeated challenges of a sublethal dose of E. coli and its relation to the hepatic energy charge.

We produced a septic model of rats in which lung water accumulation was developed. The degree of lung water accumulation was then compared with the hepatic energy status because the liver is not only a metabolic central organ but also one of the central organs of the reticulo-endothelial system (RES), and clinically, lung edema in sepsis seems to relate to failure of the RES. Three experimental models were examined to form lung water accumulation, namely: the lethal model, given E. coli 6.0 X 10(6) CFU/g BW, the sublethal model, given E. coli 2.0 X 10(6) CFU/g BW, and the repeated sublethal dose injection model, given E. coli 2.0 X 10(6) X 2 at 12 hour intervals. In the lethal models, the energy charge (EC) of the liver decreased markedly (p less than 0.001) without recovery and the lung water accumulated (p less than 0.05). In the sublethal models, EC decreased transiently (p less than 0.05) and the lung water did not increase. However, when the microbial challenge with a sublethal dose was repeated, the decreases in EC were prolonged and the lung water increased significantly after the second injection (p less than 0.001) despite a 4.9 per cent mortality during the subsequent 24 hours. It is suggested that when the decrease in liver EC is prolonged, even if it is not fatal, an accumulation of lung water is possible in septic states. This finding may help further analyses of the interrelationship between the lung and the liver in severely infected patients.

Animals↗
PubMed1991

Bile flow analysis by hepatobiliary scintigraphy in the terminal bile duct in patients with congenital malformations of the pancreatico-biliary ductal system.

The authors employed hepatobiliary scintigraphy (HBS), to study a dynamic state of bile stasis in the common bile duct and bile flow through the papilla of Vater in patients with congenital malformations of the pancreatico-biliary ductal system, such as congenital cystic dilatation of the common bile duct (CCDB) and anomalous arrangement of the pancreatico-biliary ductal system (AAPB). The HBS data were evaluated by a cholestatic index (CI), as an indicator of bile stasis in the common bile duct, and passage time (PT), as an indicator of bile flow through the papilla of Vater. CI was significantly higher in CCDB (3.67 +/- 0.92), in AAPB (2.87 +/- 0.28) and in both CCDB and AAPB with symptoms (2.86 +/- 0.59), compared with patients without CCDB or AAPB (0.96 +/- 0.41). PT was prolonged only in patients with AAPB with/without CCDB (greater than 30 minutes) compared to patients without AAPB (less than 12 minutes). This result suggested that bile stasis in the common bile duct was caused by a pooling of bile in the dilated common bile duct in CCDB, and by a disturbance of bile flow through the papilla of Vater in AAPB.

Aspartate Aminotransferases↗