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T Imanari

Publications and source records attributed to T Imanari.

At least 73 records · Page 4Linked to original sources

Chemiluminescence high-performance liquid chromatography for the determination of hyaluronic acid, chondroitin sulphate and dermatan sulphate.

A sensitive chemiluminescence high-performance liquid chromatographic method has been developed for the determination of hyaluronic acid, chondroitin sulphate and dermatan sulphate as their unsaturated disaccharide-dansylhydrazine derivatives involving an effective sample clean-up system. The dansylhydrazones of the unsaturated disaccharides derived from the hyaluronic acid, chondroitin sulphate and dermatan sulphate by chondroitinase ABC and/or chondroitinase ACII, were separated by reversed-phase chromatography using a mixture of 0.1 M sodium acetate buffer (pH 6.0) and 80% acetonitrile on a column (250 mm x 4.0 mm I.D.) packed with amide-80 silica beads (5 microns diameter). For post-column elution in the chemiluminescence system, 1 mM bis[2-(3,6,9-trioxadecanyloxycarbonyl)-4-nitrophenyl]oxalate and 3mM hydrogen peroxide in acetonitrile were used. The detection limit of each glycosaminoglycan was 100 fmol. The method was applicable to the determination of the levels of hyaluronic acid, chondroitin sulphate and dermatan sulphate in rat peritoneal mast cells.

Animals↗

Study of the measurement of chondroitin sulphates in rabbit plasma and serum.

A highly sensitive high-performance liquid chromatography method, which was established by us for the determination of chondroitin sulphates in biological substances as their unsaturated disaccharides, was applied to elucidate the qualitative and quantitative differences in chondroitin sulphates in rabbit plasma and serum samples. In this work, it was found that rabbit plasma contains low-sulphated chondroitin 4-sulphate (approximately 40% sulphation at the 4-position of N-acetyl galactosamine), while serum contains the low-sulphated chondroitin 4-sulphate and fully sulphated chondroitin 4-sulphate (approximately 96% sulphation). The latter was released from platelets during coagulation of blood.

Animals↗

Prognostic significance of location of lymph node metastases in colorectal cancer.

The prognostic value of stage of lymph node metastases was evaluated in 357 patients who underwent curative resection for colorectal cancer. Subdivision of Dukes C patients according to the number of positive nodes revealed that the five-year disease-free survival rate (5DFS) was 63 percent in the patients with one to three nodes and 53 percent in those with four or more nodes (not significantly different). Classification according to the location revealed that 5DFS was 70 percent in those who had only local node metastases (n1+), compared with 40 percent in those who had distant node metastases along the major vessels (n2+) (P < 0.001). Twelve of 38 n2+ patients had only one distant node metastasis with no local node involvement (skip metastasis). They had lower 5DFS than the n1+ patients who had three or more positive local nodes (35 percent vs. 57 percent). We conclude that the location, rather than the number, of nodal metastases has a higher impact on prognosis in colorectal cancer patients.

Colorectal Neoplasms↗

Study on proteoglycans having low-sulfated chondroitin 4-sulfate in human urine and serum.

State analysis of low-sulfated chondroitin 4-sulfate (LSC) in human urine and serum was performed by the use of high performance liquid chromatography and Western blot analysis. It was revealed that the most amount of LSC in urine is present as urinary trypsin inhibitor and a small amount (about 10% of total LSC) is as an LSC chain. The LSC in serum is mainly present as a proteoglycan such as inter-alpha-trypsin inhibitor (ITI), with a molecular weight of 212 kDa, but a small amount of LSC-proteoglycans having molecular weights of 128 and 38 kDa were also observed on SDS-PAGE. Those two compounds may be fragments of ITI, or one of the compounds (128 kDa) may be pre-alpha-trypsin inhibitor which was found by Enghild et al. (J. Biol. Chem., 264, 15975 (1989)).

Blotting, Western↗

Structural analysis of the N-linked oligosaccharides from human urinary trypsin inhibitor.

The N-linked oligosaccharides from human urinary trypsin inhibitor were purified and their structures were investigated by compositional analysis, the two-dimensional sugar map method and 500 MHz 1H-NMR. The results revealed that they were composed of disialosyl, monosialosyl and asialosyl oligosaccharides, which have the common biantennary core structure; Gal1-4GlcNAc1-2Man1-3(Gal1-4GlcNAc1-2Man1-6)M an1-4GlcNAc1-4GlcNAc.

Carbohydrate Conformation↗

Determination of human urinary hyaluronic acid, chondroitin sulphate and dermatan sulphate as their unsaturated disaccharides by high-performance liquid chromatography.

A method for the determination of hyaluronic acid (HA), chondroitin sulphate (CS) and dermatan sulphate (DS) was developed. HA, CS and DS were converted to the corresponding unsaturated disaccharides by digestion with chondroitinase ABC and/or chondroitinase AC-II and determined by high-performance liquid chromatography with fluorimetric detection using 2-cyanoacetamide as a post-column derivatization reagent. The calibration graphs for the unsaturated disaccharides were linear over the range 2 ng - 2 micrograms for each unsaturated disaccharide. This method was applied to the analysis of normal human urine.

Chondroitin Sulfates↗

Microdetermination of hyaluronic acid in human urine by high performance liquid chromatography.

A high performance liquid chromatographic (HPLC) system is described for determination of the unsaturated disaccharide (delta Di-HA) derived from hyaluronic acid (HA) in human urine by digestion with hyaluronidase SD. The effects of eluents on the separation of delta Di-HA and delta Di-0S, which is derived from the reaction of chondroitin with the enzyme, have been studied. The established chromatographic conditions were as follows--column: a stainless steel tube (4 mm i.d. x 250 mm) packed with TSKgel NH2-60; eluent: a mixture of acetonitrile and 0.1 M Tris-HCl buffer containing 0.1 M boric acid and 10 mM sodium sulphate, pH 7.0 (64:36, v/v). The strong fluorescence of unsaturated disaccharide after the reaction with 2-cyanoacetamide in alkaline medium was used for post-column detection. The calibration curve for delta Di-HA was linear in the range 5 pmol-5nmol with a practical detection limit of 2 pmol. The assay coefficients of variation (n = 5) at 200 pmol for delta Di-HA and delta Di-0S were 1.7 and 1.5%, respectively. This HPLC system has been applied to the determination of HA in human urine.

Adult↗

Analyses of rabbit platelet proteo-chondroitin sulfates localized on the surface and in the granules.

Platelet proteo-chondroitin sulfates localize on the surface and in the granules. Western blotting method is able to selectively detect core-proteins prepared from proteo-chondroitin sulfates by chondroitinase ABC digestion. A very highly sensitive high-performance liquid chromatographic method was established for the determination of chondroitin sulfates as their unsaturated disaccharides. These methods were applied to rabbit platelet proteo-chondroitin sulfates. It was found that both rabbit platelet surface proteo-chondroitin sulfate and granule proteo-chondroitin sulfate consist of a predominantly homogeneous chondroitin 4-sulfate and a single core-protein having a molecular weight of approximately 26 kDa.

Animals↗

[Metabolic study on chondroitin sulfates in rabbits].

The metabolic studies on three types of chondroitin sulfates in rabbits were performed. The analyses of chondroitin sulfates gave the following results concerning their metabolic behavior. 1) Desulfation of chondroitin sulfates on GalNAc C6 position was observed more clearly after the administration of low molecular weight chondroitin sulfate (molecular weight, 6000 and 16,000) than after that of high molecular weight one (molecular weight, 50,000). 2) The disappearance velocities of the administered low molecular weight chondroitin sulfates in the blood and those of excretion into the urine were faster than those of high molecular weight one. 3) Intact chondroitin sulfates were excreted into the urine after the administration of low molecular weight chondroitin sulfates, while chondroitin sulfates having more than 30000 molecular weight were not detected in the collected urine after the administration of high molecular weight one.

Animals↗

[Multivariate analysis for the prediction of recurrence in colorectal cancer].

We developed a predictive scale for cancer recurrence by conducting a multivariate analysis (Hayashi's discriminant analysis of qualitative data) on clinicopathologic indicators in 207 patients with Dukes' B or C colorectal cancer who underwent curative resection at our institution. Seven indicators were found to have prognostic value. Nodal status, which was divided into five categories according to the level and number of positive nodes, had the highest coefficient as a predictor of recurrence. The correlation ratio between the group showing recurrence and that showing non-recurrence was 0.49, with 74% discrimination success rate. Liver or lung metastases were more accurately discriminated than local or peritoneal failure. We assessed the accuracy of this predictive scale in 110 patients with Dukes' B or C colorectal cancer who had been surgically treated at another institution (National Cancer Center, Tokyo), and determined it to be 68%. We think that this scale using multivariate analysis for the prediction of cancer recurrence has significant clinical application in the selection of more effective postoperative adjuvant chemotherapy. It is concluded that the present predictive scale is clinically useful, but that the inclusion of other indicators such as immunohistological ones is required to increase its predictive precision.

Colorectal Neoplasms↗

Determination of citrulline and homocitrulline by high-performance liquid chromatography with post-column derivatization.

A high-performance liquid chromatographic method was developed for the determination of citrulline and homocitrulline using a post-column colorimetric reaction with o-phthaladehyde and N-(1-naphthyl)-ethylenediamine. Citrulline and homocitrulline were determined with no interferences from protein amino acids. The results show that the level of citrulline in the plasma of patients with uremia on intermittent hemodialysis is higher than that in healthy human plasma, and that homocitrulline is excreted into the urine of healthy adults.

Adult↗

State analysis of endogenous cyanate ion in human plasma.

Cyanate ion is continuously produced by spontaneous degradation of urea both in healthy human blood and in blood of patients with uremia on intermittent hemodialysis. The cyanate ion is trapped irreversibly by amino groups as N-carbamyl groups and reversibly by sulfhydryl groups as S-carbamyl groups. Consequently, these reactions keep the cyanate ion levels in plasma extremely low. Plasma proteins may play an important role in the detoxication of cyanate ion.

Adult↗

Metabolic pathways of sodium bisulfite injected intravenously in rabbits.

The metabolites of sodium bisulfite in rabbits through intravenous injection were examined according to the high performance liquid chromatography method for determination of sulfur-containing substances. Most of the administered sulfite was oxidized to sulfate, and a small part was converted to thiosulfate, S-sulfoalbumin, S-sulfoglutathione and S-sulfocysteine. Furthermore, it was found that S-sulfocysteine administered intravenously to a rabbit was partially changed to inorganic sulfate and thiosulfate. These metabolites produced from sulfite in rabbits indicated the presence of the many and complicated metabolic pathways of sulfite in vivo.

Animals↗

Profile analysis of chondroitin sulfates in human urine and serum.

A highly sensitive high-performance liquid chromatographic method with fluorometric postcolumn labeling using 2-cyanoacetamide was developed for the profile analysis of chondroitin sulfates (ChS) in normal human urine and serum. Over-sulfated disaccharide units such as di- or trisulfated unsaturated disaccharides in urine were estimated and unsaturated 6-sulfated disaccharide (delta Di-6S) was found as a major component from ChS in urine, although only small amounts of delta Di-6S from ChS were present in serum.

Chondroitin↗

Studies on metabolic pathways of cyanate in rats.

Metabolic pathways of cyanate in rats were studied by means of measurements of cyanate, carbamyl phosphate and S-carbamyl group. Approximately 30-50% of cyanate administered to rats (0.5 mmol/kg body weight) was found in buffered gastric contents, and was also detected as ammonia liberated by acid hydrolysis. However, the gastric excretion of cyanate was a temporary phenomenon just after cyanate administration. Biliary and urinary excretion of cyanate and acid-soluble S-carbamyl group are minor metabolic pathways.

Animals↗