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Biomedical subjects

I Ishizuka

Publications and source records attributed to I Ishizuka.

18 recordsLinked to original sources
PubMed1992

Substrate specificity and distribution of UDP-GalNAc:sialylparagloboside N-acetylgalactosaminyltransferase in the human stomach.

The detailed substrate specificity of the UDP-GalNAc:sialylparagloboside N-acetylgalactosaminyltransferase to form the Sd(a+) blood group active carbohydrate determinant GalNAc beta 1-4(NeuAc alpha 2-3)Gal was studied using a membrane fraction prepared from human gastric fundic mucosa. Various sialosylated oligosaccharides and gangliosides were examined as acceptor substrates. Oligosaccharide substrates were fluorescence-labelled with 2-aminopyridine, and the transferase activity was quantified by h.p.l.c. using a reversed-phase column. The structures of the products were determined by glycosidase degradation and proton n.m.r. 3'-Sialyl-lactose (II3NeuAcLac), 3'-sialyl-lactotetraose (IV3NeuAcLc4), and 3'-sialyl-lactoneotetraose (IV3NeuAcnLc4) were good substrates for the beta 1-4GalNAc transferase in gastric fundic mucosa, but 6'-sialyl-lactoneotetraose (IV6NeuAcnLc4) or 6'-sialyl-lactose (II6NeuAcLac) were not. Gangliosides with a terminal NeuAc alpha 2-3Gal residue such as GM3, sialylparagloboside, GM1b and GD1a were also studied. The activity of beta 1-4GalNAc transfer to sialylparagloboside was much higher than that to GM2, GM1b or GD1a in spite of them having the same terminal residue. Measurement of the activity of the beta 1-4GalNAc transferase in biopsy specimens demonstrated that the activity was localized in gastric fundic mucosa and was absent in pyloric mucosa, intestinal metaplasia and gastric cancer tissue. Thus the beta 1-4GalNAc transferase present specifically in fundic mucosa required a NeuAc alpha 2-3Gal residue connected to either type-1-chain or type-2-chain oligosaccharides. In glycolipids, the acceptor specificity was restricted to NeuAc alpha 2-3Gal beta 1-4GlcNAc because the NeuAc alpha 2-3Gal beta 1-3GalNAc structure in ganglio-series glycolipids was not a good acceptor substrate.

Aminopyridines↗
PubMed1992

Isolation and structural characterization of a mono-sulfated isoglobotetraosylceramide, the first sulfoglycosphingolipid of the isoglobo-series, from rat kidney.

A novel sulfoglycosphingolipid based on the isoglobo-series core structure was isolated from rat kidney and purified by column chromatographies with DEAE-Sephadex and silica beads. The structure was characterized by solvolysis, compositional analysis, proton NMR spectroscopy, Fourier-transform infrared spectroscopy, methylation analysis and liquid secondary ion mass spectrometry (LSIMS). The characteristic fragment ions for a sulfate and a sulfated N-acetylhexosamine were observed in LSIMS spectra. The two-dimensional chemical-shift-correlated spectroscopy (COSY) and nuclear Overhauser enhancement spectroscopy experiments evidenced the presence of a 3-O-sulfated N-acetylgalactosamine and a Gal alpha 1-3Gal structure in the molecule. The major ceramide consisted of 4-hydroxysphinganine linked to a C24 nonhydroxy fatty acid, deduced from both compositional analysis and LSIMS. From the above results, the following structure was established for this glycolipid: HSO3-3GalNAc beta 1-3Gal alpha 1-3Gal beta 1-4Glc beta 1-1Cer, isoglobotetraosylceramide (iGb4Cer) IV3-sulfate. Rat kidney also contained globotetraosylceramide (Gb4Cer) IV3-sulfate which has a carbohydrate core identical to that from human kidney. The yields of iGb4Cer IV3-sulfate and Gb4Cer IV3-sulfate were 0.27 and 0.07 nmol/g wet tissue, respectively.

Animals↗
PubMed1992

Sperm immobilizing antibodies react to the 3-O-sulfated galactose residue of seminolipid on human sperm.

It is well known that very few women who possess sperm immobilizing antibodies in their sera can conceive naturally even though there are no abnormalities in their reproductive organs on routine medical examination. A monoclonal antibody (MAb), designated 2H12, was produced by immunizing a BALB/c mouse with the human choriocarcinoma cell line JEG-3. MAb 2H12 showed strong sperm immobilizing activities and reacted to sulfatide and seminolipids. The sperm immobilizing activities of 2H12 were clearly absorbed with sulfatide or seminolipid whilst several other sperm immobilizing MAbs that were made by immunization with human sperm or seminal plasma could not be absorbed with the same sulfoglycolipids. The sperm immobilizing antibodies in the sera of infertile women with unknown cause were also clearly absorbed with sulfatide or seminolipid. MAb 2H12-conjugated immunobeads (MAb 2H12-IMBs) bound to motile sperm. This binding of 2H12-IMBs to sperm was competitively inhibited either by 2H12 or women's sera containing sperm immobilizing antibodies, but not by normal women's sera or several other sperm immobilizing MAbs to human sperm. These results suggest that the sperm immobilizing antibody in women's sera is directed against the 3-O-sulfogalactose residue of seminolipid on the sperm membrane.

Animals↗
PubMed1992

[Evaluation of the timing principle with small priming doses of vecuronium].

The intubating conditions using the timing principle combined with small priming doses of vecuronium were evaluated in forty patients who underwent elective surgery. They were randomly assigned to one of two groups: 1) timing, 2) timing with priming. In timing group, vecuronium 0.15 mg.kg-1 was administered, and at the onset of clinical muscle weakness, thiopental 4-5 mg.kg-1 was given promptly. Sixty seconds after thiopental, patients were intubated. In the timing with priming group, vecuronium 0.005 mg.kg-1 was administered as priming doses. Four minutes later vecuronium 0.15 mg.kg-1 was given. The administration of thiopental and the intubation were done in the same way as in timing group. The time to onset of clinical weakness after the administration of vecuronium 0.15 mg.kg-1 was significantly shorter in the timing with priming group than that in the timing group (46.1 +/- 4.8 vs. 57.6 +/- 7.8, P < 0.01). There were no significant differences in intubating score, T1, TR, onset time, and duration between the two groups. We conclude that the timing principle combined with small priming doses of vecuronium might be safe and useful for rapid tracheal intubation.

Adult↗
PubMed1979

Enzymatic sulfation of galactosyl- and lactosylceramides in cell lines derived from renal tubules.

1. The renal cell lines, JTC-12 and MDCK, not only synthesize galactosylceramide 3-sulfate and lactosylceramide 3'-sulfate in vivo, but also contain enzymes that catalyze the transfer of sulfate to galactosylceramide and lactosylceramide in vitro. 2. Concentration of cations necessary for maximum sulfotransferase activity occurred at 40 mM Ca2+ with galactosylceramide and 15 mM Ca2+ with lactosylceramide as the substrate. Na+ was also found to stimulate the sulfation of galactosylceramide, but was slightly inhibitory for the sulfation of lactosylceramide. 3. The products of the in vitro assay mixture were characterized as galactosylceramide 3-sulfate and lactosylceramide 3'-sulfate by a variety of TLC separations. 4. The apparent Km of JTC-12 cells for galactosylceramide was 17 microM, while that for lactosylceramide was 82 microM. The Km values of MDCK cells were comparable to those of JTC-12 cells. Competition studies suggested that galactosylceramide and lactosylceramide were sulfated by a single enzyme in both cell lines.

Adenosine Triphosphate↗
PubMed1978

Hormone-specific responses and biosynthesis of sulfolipids in cell lines derived from mammalian kidney.

The established cell lines isolated from mammalian kidney were characterized by its receptor activities against hormones and the ability to synthesize sulfolipids localized in the renal tubule. The level of 3':5'-cyclic AMP in JTC-12.P3 (monkey kidney) cells increased in 2 min as much as 2.5-5-fold on activation with 1.0 unit/ml of bovine parathyroid hormone or 1.9 units/ml of synthetic parathyroid hormone (1-34) resulting in intracellular cyclic AMP concentration of more than 40 pmol/mg protein. Prostaglandin E1 (14 micronM) and isopropylnorepinephrine (10 micronM) were also found to increase the concentration of cyclic AMP by more than 30- and 9-fold, respectively. Addition in medium of calcitonin, arginine vasopressin, adrenocorticotropic hormone and glucagon caused no significant changes of cyclic AMP level in the cell. In contrast, MDCK, a cell line isolated from canine kidney, reacted to arginine vasopressin, isopropylnorepinephrine and prostaglandin E1 and only slightly to parathyroid hormone. MDBK cell line derived from bovine kidney or fibroblast cell lines from rat lung and guinea pig kidney did not react to any of the hormones specific to kidney, i.e. arginine vasopressin, calcitonin or parathyroid hormone in the presence of theophylline. However, in the presence of 2 mM isobutylmethylxanthine, small but significant elevation of cellular cyclic AMP levels in response to calcitonin, arginine vasopressin, isopropylnorepinephrine and prostaglandin E1 was observed. The cell lines JTC-12, MDCK and MDBK, when incubated with H235SO4, incorporated the isotope into sulfolipids assigned as sulfatides and ceramide dihexoside sulfate or in MDCK also into cholesterol sulfate. The results suggested that JTC-12, MDCK and MDBK cell lines are epithelial origin and also JTC-12 and MDCK originated most probably from renal tubular cells of cortex and medulla, respectively.

Calcitonin↗
PubMed1977

Glycolipid composition of human testis at different ages and the stereochemical configuration of seminolipid.

1. The sterochemical configuration of glycerol moiety of seminolipid (1-O-alkyl-2-O-beta-D-(3'-sulfo) galactopyranosylglycerol: Ishizuka, I., Suzuki, A. and Yamakawa, T. (1973). J. Biochem. 73, 77--87) from mammalian testis was determined by measurement of optical rotatory dispersion of alkylglyceryl ether moiety, establishing the final structure of the major glycolipid from mature testis of mammals as 1-O-alkyl-2-O-beta-D-(3'-sulfo)-galactopyranosyl-sn-glycerol. 2. The simple and sensitive modification of fluorometric determination of galactolipids and sphingolipids was described. This method does not require lipid extraction from silica gel of thin-layer chromatography, and allows a quantitative assessment of the sphingoglycolipids and galactosyl glycerides using a small amount of testicular tissue. 3. Various lipid classes from human testis of varying age groups were compared. The total lipid and seminolipid content was highest in adult (40-years-old) testis amounting to 39.0 mg and 158.7 nmol/g tissue, respectively. Seminolipid was not detected in infant (2 years old) or child (9 years old) and found only at the concentration of 25.3 nmol/g in the testis of aged (60--90 years of age) supporting the postulated relation of seminolipid with spermatogenesis and sexual activity. In contrast, ganglioside content was highest in the testis of aged probably reflecting fibrosis of the testis. The main ganglioside was found to be hematoside with N-acetylneuraminic acid. Gangliosides Gm1 and GD1a were also detected.

Adult↗
PubMed1977

Micro-scale determination of seminolipid by high performance liquid chromatography and its application for determination of the seminolipid content in boar spermatozoa.

Seminolipid (1-O-alkyl-2-O-acyl-beta-3'sulfogalactosyl glycerol) was detected on a micro-scale by high performance liquid chromatography (HPLC) using a UV detector. The benzoylation method of McCluer and Evans ((1973) J. Lipid Res. 14, 611-617) was used to convert seminolipid to a derivative with UV absorption. Total lipid from germinal tissue was benzoylated and the product was separated by Florisil and Sephadex LH-20 column chromatographies. The seminolipid obtained in the effluent from the Sephadex LH-20 column was determined by HPLC. Eighty percent recovery of 35S-labelled seminolipid was obtained by this method. The method was used to determine the seminolipid content of boar spermatozoa: 980 nmoles of seminolipid was found per gram of packed cells.

Animals↗
PubMed1976

Sulphogalactolipid sulphohydrolase activity of arylsulphatase purified from a marine gastropod Charonia lampas.

Sulphatide, cerebroside 3-sulphate was hydrolyzed at a considerable rate by arylsulphatase (aryl-sulphate sulphohydrolase, EC 3.1.6.1) purified from a marine gastropod, Charonia lampas. However, it was scarcely hydrolyzed by glycosulphatase (sugar-sulphate sulphohydrolase, EC 3.1.6.3) from the same origin. The same was observed with seminolipid, a sulphoglycerogalactolipid. The enzymatic characteristics of both sulphogalactolipid and sulphohydrolase activities of the arylsulphatase were determined as follows. The enzyme activities are stimulated by the addition of sodium taurodeoxycholate and MnCl2. The pH optimum of sulphatide sulphohydrolase activity was pH 5.0, while seminolipid sulphohydrolase activity had maximum activity at pH 5.5. Both of these pH versus activity curves were broad. The Km value was 6.22-10-5 M for both substrates. However, the V values were sulphatide were lower by a factor of one-third than those with seminolipid. These enzyme activities were inhibited by substrates of the arysulphatase, i.e., p-nitrophenyl sulphate, p-nitrocatechol sulphate, ascorbate 2-sulphate and each other sulphogalactolipid, but not by glucose 6-sulphate. Sulphate and phosphate anions inhibited both of the enzyme activities.

Animals↗
PubMed1975

Heterogeneity of carcinoembryonic antigen. II. Monosaccharide composition of concanavalin A-reactive and -nonreactive CEA.

Monosaccharide compositions of Con A-reactive CEA (CEA-M) and Con A-nonreactive CEA (CEA-P) separated from two different samples of CEA were analysed by gas liquid chromatography. It was revealed that all CEA subfractions possessed N-acetylglucosamine, fucose, and galactose residues. One out of 4 subfractions did not contain sialic acid and another one lacked glucose in its carbohydrate moiety. N-acetylgalactosamine was not detected in measurable amount in any of the 4 subfractions. A large amount of mannose was found in CEA-M, but only a small amount in CEA-P.

Acetylgalactosamine↗
PubMed1975

Isolation and characterization of a ganglioside containing fucose from boar testis.

A ganglioside containing fucose (fucoganglioside) was obtained from boar testis and purified by silicic acid and DEAE-cellulose column chromatographies and preparative thin layer chromatography. The structure of this ganglioside, determined by chemical and enzymatic methods was: (see article). Its fatty acids were mainly long chain saturated ones (20 : 0, 22 : 0, 24 : 0). Its long chain bases consisted of 27% C(16:1) sphingosine and 68% C(18:1) sphingosine.

Animals↗
PubMed1975

Glycolipids of the fish testis.

Glycolipids were purified from the total lipid extract of the testis or milt of a kind of puffer (Fugu rubripes rubripes) by adsorption column chromatography using silicic acid and magnesium silicate and by preparative silica gel TLC. The glycolipids were identified as glucosylceramide (116 mug/g wet tissue) and galactosylceramide 26.7 mug/g). Seminolipid, a sulfagalactolipid specific to mammalian testis was not detected, but the presence of a small amount of sulfatide (15.2 mug/g) was demonstrated. The long-chain bases of both cerebrosides were mainly C18-sphingenine, but in sulfatide, C20-sphingenine was more abundant than C18-sphingenine. In both cerebrosides and sulfatide, the fatty acid compositions were similar, with nervonic acid as the predominant component. Two species of gangliosides were also obtained and were identified as N-acetylgalactosaminyl(1 leads to 4)[N-acetylneuraminyl(2 leads to 3)]galactosyl(1leads to 4)glucosylceramide (59.8 mug/g) and N-acetylneuraminyl(2 leads to 3)galactosyl(1 leads to 4)N-acetylglucosaminyl(1 leads to 3)galactosyl(1 leads to 4)glucosylceramide (45.0 mug/g). The long-chain bases of the two gangliosides consisted of C18-spingenine and C20-sphingenine, and the major fatty acids were palmitic and stearic acids.

Animals↗