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K Nohara

Publications and source records attributed to K Nohara.

At least 37 records · Page 2Linked to original sources

Transcapillary ultrafiltration and peritoneal equilibration test in pediatric patients.

The usefulness of the peritoneal equilibration test (PET) in children is unknown. The relationship between transcapillary ultrafiltration and PET was investigated in order to evaluate the usefulness of PET in children. PET was performed on 14 patients undergoing peritoneal dialysis. Their age and bodyweight ranged from 3.8 to 23.6 years and 10.2 to 55.8 kg, respectively. The patients were divided into two groups according to bodyweight; group A patients weighed < or = 40 kg (n = 7) and group B patients weighed > 40 kg (n = 7). There was no significant difference in the mean infusion volume per bodyweight between the two groups, but the mean infusion volume per body surface area was smaller in group A than in group B. Group A showed a more rapid equilibration of dialysate glucose and creatinine than group B. Higher normalized mass transfer area coefficients were evident in group A. In spite of the lower effective glucose gradient in group A, the transcapillary ultrafiltration capacity (TUFC) showed no difference between the two groups. When the slope indices of the regression equations between the two groups were compared, the slopes of the regression in the relationship between TUFC and dialysate (D) ratios D/Do glucose or D/P creatinine in group A were steeper than those in group B. Results of the present study indicate that the larger peritoneal area to infusion volume in patients with smaller body size results in both a rapid equilibration of solutes and sufficient transcapillary ultrafiltration.

Adolescent↗

Ganglioside GM3 inhibits interleukin-3-dependent bone marrow-derived mast cell proliferation.

To investigate the modulated proliferation of an interleukin-3(IL-3)-dependent cell by exogenous ganglioside GM3, mouse bone marrow-derived mast cells (BMMC) were cultured with various concentrations of GM3 in the presence of IL-3. By 4 weeks of culture, most of the nonadherent cells were alcian blue-positive mast cells. Culturing 2-week-cultured BMMC with GM3 for 1 week reduced the number of alcian blue-positive cells, but the increased total histamine content of BMMC was observed. To examine the effect of GM3 on the synergistic response by IL-3 and interleukin-4 (IL-4), 3-week-cultured BMMC were cultured with GM3 in the presence of IL-3 and IL-4 for 1 week. Although the addition of IL-4 to culture medium increased the number of BMMC, treatment with GM3 reduced its proliferative activity. Concerning the effect of GM3 on cell membrane, there are no changes in the expression of IgE receptors on BMMC treated with GM3 though a low concentration of GM3 increased it. However, the production of tumor necrosis factor-alpha from BMMC treated with GM3 was significantly suppressed. These results indicate that in vitro treatment with exogenous GM3 inhibited the proliferative response of IL-3-dependent mast cell populations and modulated its characteristics.

Animals↗

Glycosphingolipids of rat T cells. Predominance of asialo-GM1 and GD1c.

Glycosphingolipids play an important role in the immune response, yet their compositions in T and B cells which mediate cellular and humoral immunity, respectively, have not been elucidated. In this study, characteristic features of glycosphingolipids in rat T lineage cells were revealed by comparing the gangliosides and neutral glycolipids of spleen T- and beta-cell-enriched fractions and thymocytes. In T cells, GD1c(NeuGc,NeuGc), a unique ganglioside synthesized through asialo-GM1 (GA1), was the predominant ganglioside as previously found in thymocytes [Nohara, K., Suzuki, M., Inagaki, F., & Kaya, K. (1991) J. Biochem. (Tokyo) 110, 274-278], and the amount was much higher than in thymocytes. In addition, three other GA1-derived gangliosides were detected in T cells and identified as GM1b(NeuAc), GM1b(NeuGc), and GD1 alpha(NeuAc,NeuAc). In contrast, GD1 alpha(NeuAc,NeuAc) was not discernible in thymocytes, although gangliosides corresponding to GM1b(NeuAc) and GM1b(NeuGc) were detected. The neutral glycolipids of T cells contained almost exclusively GA1, while thymocytes contained much lower amounts. The predominance of these GA1-derived gangliosides was confirmed as a singular feature of T lineage cells by comparison with gangliosides of spleen B-cell-enriched fractions which mainly consisted of gangliosides synthesized through GM3 and GM1. Furthermore, the unique structures, which contain the GM1 core and the extended modification of the lacto series, alpha Gal-LacNAc-GM1, alpha Gal-(LacNAc)2-GM1, and sialyl-LacNAc-GM1, were found in B-cell-enriched fractions. Unexpectedly, the neutral glycolipid composition of the thymocytes resembled that of the B-cell enriched fraction rather than that of the T cells.

Animals↗

An activation-associated ganglioside in rat thymocytes.

During the activation of rat thymocytes elicited by stimulation with 12-O-tetradecanoylphorbol-13-acetate (TPA) and calcium ionophore A23187, the total amount of gangliosides per cell was demonstrated to increase, reaching a maximum level several times higher than that of resting cells at 48 h after activation when measured at 24-h intervals. The amount of the overwhelmingly predominant ganglioside in resting thymocytes, GD1c (NeuGc,NeuGc) (Nohara, K., Suzuki, M., Inagaki, F., and Kaya, K. (1991) J. Biochem. (Tokyo) 110, 274-278), was found to increase further as a result of activation. Furthermore, another ganglioside, which was barely recognizable in resting thymocytes, was found to increase in activated and proliferating thymocytes to a level similar to that of GD1c. This activation-associated ganglioside was isolated and its structure examined. On the basis of the results of compositional analysis, methylation analysis, sialidase hydrolysis followed by detection with cholera toxin B subunit on TLC, and proton NMR spectroscopy, this ganglioside was clarified to be a rare species of GD1b containing two N-glycolylneuraminic acid residues. On the other hand, when the thymocytes were activated using concanavalin A (ConA) as a stimulant, the amount of gangliosides per cell was increased more strikingly than that in thymocytes activated with TPA and A23187. In the ConA-activated thymocytes, many other gangliosides, in addition to GD1c and GD1b-(NeuGc,NeuGc), were demonstrated to appear in large amounts. The cause of this difference in gangliosides between thymocytes activated with a combination of TPA and A23187 and those activated with ConA is also discussed.

Animals↗

A novel disialoganglioside in rat spleen lymphocytes.

A novel ganglioside has been identified as the predominant disialoganglioside of the lymphocytes prepared from rat spleen. The ganglioside was isolated from rat spleen and characterized by compositional analysis, methylation analysis, sialidase hydrolysis, proton NMR spectroscopy, and negative ion fast atom bombardment mass spectrometry. The structure was determined as follows. [formula: see text] This ganglioside is a unique derivative of N-acetyllactosaminyl-GM1. The three monosialogangliosides containing N-acetyllactosaminyl-GM1 structure, which had been originally isolated from rat spleen (Nohara, K., Suzuki, M., Inagaki, F., Ito, H., and Kaya, K. (1990) J. Biol. Chem. 265, 14335-14339), were also found in the lymphocytes and were hardly detected in the spleen remnant tissue depleted of single cells. On the other hand, GD1c(NeuGc,NeuGc) (IV3(NeuGc alpha 2-8NeuGc)-Gg4Cer), the overwhelmingly predominant ganglioside of rat thymocytes (Nohara, K., Suzuki, M., Inagaki, F., and Kaya, K. (1991) J. Biochem. (Tokyo) 110, 274-278), was demonstrated to be only a minor component of the gangliosides of the spleen lymphocytes. These results suggested that GD1c is characteristic for the immature T lineage lymphoid cells and the gangliosides having lactosaminyl-GM1 structure are specific for other populations of the lymphocytes in rat.

Animals↗

Purification and characterization of secretory proteinase of Candida albicans.

Proteolytic activity of medically important yeasts was tested in both YCB-BSA agar and medium. All of 134 strains of Candida albicans, 13 of 18 strains of Candida tropicalis and 11 of 18 strains of Candida parapsilosis had this activity, while none of 52 Candida glabrata strains or of 11 Cryptococcus neoformans strains tested had proteolytic activity. Strains of C. albicans fell into five groups based on the level and time-course of in vitro proteinase productivity. Five strains randomly selected from each group were tested for pathogenicity in mice. The strain possessing the strongest pathogenicity was used to purify proteinase. The molecular weight of the proteinase was approximately 44,000 daltons and its isoelectric point was pH 4.2. Optimal pH of the proteinase was 3.2 and the enzyme was stable below pH 7.0 and lost its activity above pH 8.0 at 37 C in a 60-min incubation. The 23 amino acid sequence of the proteinase N-terminus was determined.

Amino Acid Sequence↗

A GM1b-derived disialoganglioside GD1c is the predominant ganglioside of rat thymocytes.

The thin-layer chromatographic (TLC) pattern of gangliosides of rat thymocytes showed a profile characterized by the occurrence of a predominant ganglioside which did not correspond to any reference gangliosides of rat brain. The ganglioside was isolated from rat thymus, and characterized by compositional analysis, methylation analysis, sialidase treatment, negative-ion fast atom bombardment (FAB) mass spectrometry, and proton NMR spectroscopy. The structure was elucidated to be NeuGc alpha 2-8NeuGc alpha 2-3Gal beta 1-3GalNac beta 1-4Gal beta 1-4Glc beta 1-1Cer. This is the major ganglioside of rat thymus lymphoid cells and is one of the GM1b-derived gangliosides, GD1c, having two N-glycolylneuraminic acids. This is the first report on the occurrence of GD1c in normal animal cells.

Animals↗

Identification of novel gangliosides containing lactosaminyl-GM1 structure from rat spleen.

Two novel monosialogangliosides were isolated from rat spleen. The structures of the gangliosides (shown below, where NeuNGc is N-glycolylneuraminic acid) were characterized by compositional analysis, methylation analysis, hydrolyses with exoglycosidases, direct probe fast atom bombardment mass spectrometry, and proton nuclear magnetic resonance spectrometry. A novel structure common to both gangliosides was N-acetyllactosaminyl-GM1 LacN-GM1; where GM1 is II3NeuAc-GgOse4Cer), and this is the first paper to report the occurrence of a new group of gangliosides. [formula: see text] Furthermore, in a monosialoganglioside fraction of rat spleen, the occurrence of a ganglioside having two lactosamine units (Gal alpha 1-3(Gal beta 1-4GlcNAc beta 1-3)2-GM1(NeuAc) (alpha Gal-(LacN)2-GM1] was suggested. These gangliosides have a unique structure, which includes the ganglio series ganglioside core and the extended modification characteristic of the lacto series.

Animals↗

2-(7,13-Dihydroxy-2-trans-octadecenoylamino)ethanesulfonic acid (lipotaurine) as an intermediate of taurolipids biosynthesis.

A hydroxy fatty-acid-combined taurine (lipotaurine) was found in the taurolipids fraction of Tetrahymena thermophila. Lipotaurine accounted for about 1.4% of the total taurolipids of the cells, and was composed of taurine and 7,13-dihydroxy-2-trans-octadecenoic acid. By nuclear magnetic resonance, mass and infrared spectrometries, the chemical structure of lipotaurine was identified as 2-(7,13-dihydroxy-2-trans-octadecenoylamino)ethanesulfonic acid. When cells of T. thermophila were incubated with the double-labeled lipotaurine which was biosynthesized from [2(n)-3H]taurine and [1-14C]stearic acid, both the radioactivities were detected in taurolipid A, B and C. Furthermore, the ratio of the radioactivities of 3H and 14C in the lysotaurolipids were the same as that of the lipotaurine. From these results, it is suggested that lipotaurine is an intermediate of taurolipid biosynthesis.

Animals↗

A unique fucoganglioside with blood group B determinant in rat spleen.

A unique fucoganglioside was isolated from rat spleen and characterized by compositional analysis, methylation analysis, exoglycosidase treatment, negative ion fast atom bombardment (FAB) mass spectrometry, and proton NMR spectrometry. The ganglioside was identified as alpha Gal,Fuc-GM1(NeuGc), which has the blood group B determinant at the nonreducing termini, as shown below: (formula; see text) This is the first report describing the occurrence in nature of alpha Gal,Fuc-GM1 containing N-glycolylneuraminic acid.

ABO Blood-Group System↗

Effect of di-n-octyl phthalate on fatty acid composition of phosphatidylcholine in Tetrahymena.

We examined the effect of di-n-octyl phthalate (DOP) on fatty acid composition of phosphatidylcholine (PC) in Tetrahymena pyriformis NT-1. When Tetrahymena cells were grown in DOP-containing proteose peptone medium, the cell growth was repressed. This repression was attended by decreases in the PC content of the cells and decreases in oleic (18:1), linoleic (18:2) and linolenic (18:3) acids of PC and an increase in palmitoleic acid (16:1). The ratio of 18:1/stearic acid (18:0) of PC in cells grown in DOP-containing medium was lower than that of control cells, while the ratio of 16:1/palmitic acid (16:0) was higher than that of control. On the other hand, no changes in the ratios of 18:2/18:1 and 18:3/18:2 were observed. The activity of microsomal stearoyl-CoA desaturase from cells grown with DOP (0.63 mumol/ml medium) decreased to 27% of that from control cells, while the microsomal palmitoyl-CoA desaturase activity increased to 210% of the control value. By the addition of dioleoyl glyceride to the DOP-containing medium, the effects of DOP on Tetrahymena cells were completely blocked. These results suggest that the changes in fatty acid composition of PC may be due to the alteration of the substrate specificity of microsomal delta 9-desaturase, and the decrease in stearoyl-CoA desaturase activity may be a cause for the cell growth repression.

Animals↗

Correlations between physical and physiological properties of various preparations of lung surfactant.

The physical and physiological properties of natural surfactant were investigated after the addition of various synthetic lipids. Three types of surfactant were studied: 1. Bovine surfactant with rapid spreading (1.6 s) and a relatively high minimal surface tension during surface compression (16 mN/m). 2. The same surfactant enriched with dipalmitoylphosphatidylcholine (DPPC), tripalmitin, and palmitic acid showing slow spreading (55 s) and low minimal surface tension (5 mN/m). 3. The same surfactant enriched with DPPC and dipalmitin, showing rapid spreading (1.8 s) and low minimal surface tension (6 mN/m). The physiological properties of these surfactants were evaluated in immature newborn rabbits. All three preparations effectively improved lung expansion and stability in pressure-volume recordings, increased tidal volumes during artificial ventilation, and enhanced alveolar volume density in histological sections. The magnitude of the therapeutic effects was similar for non-enriched and enriched materials. Thus, wide variations in in vitro surface properties do not seem to influence the in vivo activity of the surfactant preparations.

1,2-Dipalmitoylphosphatidylcholine↗

The role of the low-molecular weight (less than or equal to 15,000 daltons) apoproteins of pulmonary surfactant.

An artificial surfactant was prepared by combining synthetic dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylglycerol and the low-molecular weight (less than or equal to 15,000 daltons) surfactant apoproteins in the proportions 80:20:5. In the Wilhelmy balance, this surfactant formed a film with an equilibrium surface tension of 29 mN/m; surface tension was reduced to nearly zero during cyclic film compression, with effective respreadability during multiple compression-expansion cycles; similar surface properties were recorded with a pulsating bubble. When instilled into the airways of artificially ventilated immature newborn rabbits, the apoprotein-based artificial surfactant produced a five-fold increase in tidal volumes at insufflation pressure 25 cm H2O; this effect is similar to that obtained in previous experiments with natural surfactant phospholipids, administered in equal concentration (5 mg/ml). Higher concentration of the apoprotein-based surfactant could not be evaluated in vivo due to the high viscosity of the material. Systematic studies should be undertaken to find out whether an even more effective artificial surfactant could be prepared from the low-molecular weight apoproteins and other combinations of synthetic phospholipids.

1,2-Dipalmitoylphosphatidylcholine↗

Studies on experimental coronary insufficiency. Effect of L-carnitine on myocardial ischemia produced by sympathetic-nerve stimulation with high plasma fatty acids.

Our previous studies revealed that sympathetic-nerve stimulation (SNSt) plays an important role in the precipitation and the augmentation of myocardial ischemia in dogs with coronary constriction. To clarify the underlying mechanism of the detrimental effect of free fatty acids (FFA) at a high plasma concentration and the beneficial effect of L-carnitine on myocardial ischemia, ischemic changes following SNSt were compared among three groups of dogs with mild or moderate coronary constriction: a saline control group, an intralipid [(IL) 0.1 ml/kg per min + heparin 5 mg/kg] group, and an IL + L-carnitine (200 mg/kg) group. High plasma concentration of FFA aggravated the ischemic changes induced by SNSt in dogs with coronary constriction, in which no signs of increase in myocardial oxygen consumption were seen. L-Carnitine clearly alleviated the mechanical dysfunction, acceleration of anaerobic metabolism, depletion of myocardial contents of high-energy phosphates, myocardial accumulation of lactate, and ECG ischemic changes that were augmented by high plasma FFA in the coronary-constricted dogs with SNSt. From these findings, it was suggested that an increased plasma FFA might aggravate myocardial ischemia, at least, produced by SNSt in dogs with mild or moderate coronary constriction and that L-carnitine might improve the ischemia augmented by FFA, presumably by reducing myocardial accumulation of FFA intermediates.

Animals↗