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Biomedical subjects

H Nomoto

Publications and source records attributed to H Nomoto.

At least 37 records · Page 2Linked to original sources
PubMed1994

Highly efficient transfection into primary cultured mouse hepatocytes by use of cation-liposomes: an application for immunization.

Transfection methods for primary cultured mouse hepatocytes were examined. Of four conventional transfection methods examined, involving use of DEAE-dextran, calcium phosphate, cation-liposomes (lipofection), and cation-multilamellar liposomes, only cation-liposomes induced highly efficient transfection into primary cultured mouse hepatocytes. The highest transfection rate reached more than 60% of the total cells. Three other commonly used cell types (CHO-K1, COS-1, 3T3-L1) were also tested as target cells, but highly efficient transfection was observed specifically in primary cultured mouse hepatocytes. The transfection remained at a high level from 6 to 48 h after the start of incubation with the cation-liposome-DNA complex in the absence of serum, and the transfection rate decreased in inverse relation to the increase in cell density. The transfection was inhibited by free low density lipoprotein (LDL), EDTA, and an endocytosis inhibitor, cytochalasin B. These data suggest that the transfection is mediated not only by membrane fusion, as is generally accepted, but also by endocytosis. This information should be useful for research in hepatocyte biology and the development of gene therapy. As one of the applications, simple and successful immunization was achieved by administration of hepatocytes transfected with murine adhesion molecule, integrin VLA beta 1 subunit, genes into a Syrian hamster.

3T3 Cells↗
PubMed1993

[A case of marked eosinophilia in peripheral blood induced by rhGM-CSF].

A 53-year-old man underwent chemotherapy (CDDP, VDS, MMC) for treatment of lung cancer. He was given 125 micrograms/m2 of GM-CSF subcutaneously every day for 8 consecutive days, in order to prevent neutropenia. Three days after starting GM-CSF therapy, marked eosinophilia in peripheral blood was observed. The maximum eosinophil count was 89% of leukocytes. Nine days after stopping the treatment with GM-CSF, the number of eosinophils had normalized spontaneously. There were no clinical symptoms except for slight fever, up to 37.5 degrees C. Moreover, there was no relationship between the number of eosinophils and the serum levels of cytokines (IL-3, IL-5, GM-CSF), although we observed minimal but significant elevation of serum ECP level. This case indicates that GM-CSF may induce marked eosinophilia rather than widely stimulating granulocytes and monocytes.

Blood Proteins↗
PubMed1992

Detailed structural analysis of asparagine-linked oligosaccharides of the nicotinic acetylcholine receptor from Torpedo californica.

The structures of the major oligosaccharide moieties of the nicotinic acetylcholine receptor (AcChoR) protein from Torpedo californica have been reported [Nomoto, H., Takahashi, N., Nagaki, Y., Endo, S., Arata, Y. and Hayashi, K. (1986) Eur. J. Biochem. 157, 233-242] to be high-mannose types. Here we report detailed analyses of the structures of the remaining oligosaccharides in this receptor. The sialylated oligosaccharides released by glycopeptidase (almond) digestion were separated according to the number of sialic acid residues using high-performance anion-exchange chromatography with pulsed amperometric detection. After removal of sialic acid from each fraction, the resulting neutral oligosaccharides were separately pyridylaminated and were analyzed by a combination of sequential exoglycosidase digestion and HPLC, then identified on a two-dimensional sugar map. The structures of two desialylated pyridylamino-oligosaccharides were further analyzed by high-resolution proton NMR. Each oligosaccharide was composed of species containing varying numbers of sialic acids. The desialylated complex-type oligosaccharides of AcChoR consisted of ten, eight and one different biantennary, triantennary and tetraantennary oligosaccharide, respectively. The biantennary oligosaccharides were divided into two groups; oligosaccharides with fucose at the proximal N-acetylglucosamine (six varieties) and oligosaccharides without fucose (four varieties). Each group consisted of species differing in the number of terminal galactose residues. The major component of the biantennary oligosaccharides had two galactose residues at the non-reducing termini. The terminal alpha-galactose residue(s) linked to C3 of beta-galactose were found in the fucose-containing biantennary oligosaccharides (two varieties). The triantennary oligosaccharides were also divided into two groups; oligosaccharides with (four varieties) and without (four varieties) besecting N-acetylglucosamine. These groups were composed of species differing in the number of terminal galactose residues. The major component of the triantennary oligosaccharides was fully galactosylated with three galactose residues. An unusual group, Gal beta 1-3GlcNAc, was present in low levels in the triantennary oligosaccharides. In contrast, the tetraantennary oligosaccharide was composed of only one species, which is fully galactosylated with four galactose residues.

Animals↗
PubMed1992

Mitogen-induced tyrosine-phosphorylated 41- and 43-kDa proteins are family members of extracellular signal-regulated kinases/microtubule-associated protein 2 kinases.

Two antipeptide antibodies, one against the peptide corresponding to residues 307-327 (alpha Y91) and one against the peptide corresponding to the C-terminal portion (alpha C92) of the deduced amino acid sequence of the extracellular signal-regulated kinase 1 (ERK1), precipitated two 41-kDa and/or two 43-kDa phospho-proteins from mitogen-stimulated Swiss 3T3 cells. Electrophoretic mobilities on two-dimensional gels of the immunoprecipitated 41- and 43-kDa phosphoproteins were similar to those of the 41- and 43-kDa cytosol proteins, whose increased tyrosine phosphorylation we and others had originally identified in various mitogen-stimulated cells (Cooper, J. A., Sefton, B. M., and Hunter, T. (1984) Mol. Cell. Biol. 4, 30-37; Kohno, M. (1985) J. Biol. Chem. 260, 1771-1779); phosphopeptide map analysis revealed that they were respectively identical molecules. All those phosphoproteins contained phosphotyrosine, and the more acidic forms contained additional phosphothreonine. Immunoprecipitated 41- and 43-kDa phosphoproteins had serine/threonine kinase activity toward myelin basic protein (MBP) and microtuble-associated protein 2 (MAP2). With the combination of two-dimensional gel electrophoresis and the kinase assay in MBP-containing polyacrylamide gels of the alpha Y91 immunoprecipitates, with or without phosphatase 2A treatment, we showed that only their acidic forms were active. These results clearly indicate that 41- and 43-kDa proteins, the increased tyrosine phosphorylation of which is rapidly and commonly induced by mitogen stimulation of fibroblasts, are family members of ERKs/MAP2 kinases and that phosphorylation both on tyrosine and threonine residues is necessary for their activation.

3T3 Cells↗
PubMed1992

Higher cortical dysfunction, antiphospholipid antibodies and neuroradiological examinations in systemic lupus erythematosus.

We performed neuropsychological tests to investigate higher cortical dysfunction in 21 patients with systemic lupus erythematosus (SLE). We also measured antiphospholipid antibodies (APA), performed brain computed tomography (CT), and obtained a single photon emission CT (SPECT) to measure regional cerebral blood flow (rCBF) in order to elucidate a possible relationship between APA and higher cortical dysfunction. Higher cortical dysfunction was noted in as many as 16 (76%) out of 21 cases. APA were positive in 8 (38%) out of 21 cases. Although the relationship between APA and higher cortical dysfunction was not significant, patients positive for lupus anticoagulant (LA) were found to have higher cortical dysfunction. Brain CT revealed at least one abnormality in 6 cases (29%) but none had a localized lesion, SPECT disclosed a reduced rCBF in 9 cases (43%). The findings on brain CT and SPECT were unrelated to higher cortical dysfunction.

Adolescent↗
PubMed1992

Prevention of aortic calcification in patients on hemodialysis by long-term administration of vitamin E.

The effects of vitamin E on the progress of atherosclerosis in patients on hemodialysis was investigated clinically using ACI. There was a significant suppression of the increase in ACI in group A, compared to group B, at the time of observation in each year. On the other hand, no significant changes were noted in BWD, CTR, BP and blood chemical examination, except that the level of MDA was significantly decreased in group A as compared with that in group B 4 years later. Since ACI is an index representing atherosclerosis, the results of this study seemed to suggest that the progress of atherosclerosis was suppressed by long-term administration of vitamin E in patients on hemodialysis.

Aortic Diseases↗
PubMed1992

Crosslinking of proteins in acetylcholine receptor-rich membranes from Torpedo californica: relation of 43-kD protein and Torpedo dystrophin to acetylcholine receptor.

We examined the spatial relation of 43-kD protein and Torpedo dystrophin, which are cytoplasmic peripheral membrane proteins in the nicotinic acetylcholine receptor (AChR)-rich membranes, to AChR. We used three kinds of the heterobifunctional crosslinking reagents to crosslink proteins in the AChR-rich membranes. Products crosslinked by SMPB (14.5 A span) including 43-kD protein and Torpedo dystrophin appeared at the tops of the stacking gels at the concentrations of 8.89 x 10(-5)M to 8.89 x 10(-3)M SMPB. High molecular weight materials (crosslinked products) increased with increasing concentrations of the crosslinker. On the other hand, band intensity of alpha, beta, and delta subunits of AChR remained unchanged up to a concentration of 2.67 x 10(-3)M SMPB, while the band of gamma subunit diminished at the same concentrations as did that of the 43-kD protein. Torpedo dystrophin was also crosslinked at the same concentrations as were effective for the 43-kD protein and gamma subunit. On the basis of these results, we conclude that the 43-kD protein is intimately associated with the gamma subunit of AChR and Torpedo dystrophin.

Animals↗
PubMed1992

Neurotoxin-binding activity in the supernatant fraction of the electric organ from Torpedo californica.

We found neurotoxin-binding activities in the supernatant fraction obtained by ultracentrifugation of a homogenate of the electric organ dissected from the electric ray, Torpedo californica. While about half of the activity was estimated as due to acetylcholine receptors in dispersed microparticles, the remainder was unassigned. A part of the latter, detected with alpha-bungarotoxin, eluted ahead of the alpha-bungarotoxin-acetylcholine receptor complex on a Sepharose CL-6B column in the presence of 1% Triton X-100. Another component eluted after this complex. Although these activities were immunologically related to AChR, they were different from AChR in their size and reactivity with Concanavalin A. We are currently seeking to characterize these toxin-binding components at present. The existence of such activities is interesting since they may possibly function in regulating signal transduction at the neuromuscular junction.

Animals↗
PubMed1992

[Clinical course of asthmatics with severe asthma attack].

This study was conducted on 39 patients whose severe attacks of bronchial asthma with disturbance of consciousness required admission to the ICU of our hospital between 1984 and 1989. Among the 39 patients, there were 16 deaths. Most patients collapsed suddenly at home and were taken to our hospital. Arterial blood gas analysis at the time of admission to the ICU revealed that the PaO2 levels were as high as 252.6 +/- 57.6 (mean +/- S.E.) Torr in non-survivors and 221.0 +/- 29.7 Torr in survivors, with no significant difference because of prior oxygen therapy in almost all cases. Systolic blood pressure was 14.8 +/- 10.8 (mean +/- S.E.) mmHg, with marked circulatory disturbance in the fatal cases. Most patients displayed marked disturbance of consciousness, but maintenance of blood pressure led to recovery without sequelae despite marked disturbance of consciousness in most patients.

Adolescent↗
PubMed1992

[A case of plasmacytoma of the ribs with intrathoracic tumors].

A 75-year-old male was admitted because of two tumors, one in the left middle lung field and one in the right upper lung field. Chest CT revealed intrathoracic tumors extending from destroyed ribs. Biopsy specimens of both tumors showed well-differentiated plasmacytoma. Retrospective investigation suggested that solitary plasmacytoma of bone (SPB) originating in the left fourth rib had developed into multiple myeloma (MM). Both tumors were treated with doses of 50 Gy irradiation and responded very well. Intrathoracic plasmacytomas have rarely been observed, so we have no established classification or therapy. According to reported cases, we classified intrathoracic plasmacytomas into 5 groups, and consider that treatment with doses of over 40 Gy irradiation was adequate for local control.

Aged↗
PubMed1991

Effect of platelet-activating factor on lipoprotein lipase and blood lipids.

We investigated the effect of platelet-activating factor (PAF) and of the PAF specific antagonist CV-6209 on plasma lipid metabolism, and particularly on post-heparin plasma lipolytic activity in male Wistar rats. Lipoprotein lipase (LPL) activity was enhanced by intravenous injection of PAF before intravenous injection of heparin when the PAF dose was low (0.2 micrograms/kg). PAF activated hepatic triacylglycerol lipase (HTGL) activity dose-dependently. Plasma triacylglycerols (TG) significantly decreased with the activation of LPL and/or HTGL. Plasma total cholesterol (TC) and phospholipid (PL) levels decreased at a low dose of PAF (0.2 micrograms/kg), but increased when higher doses were used. The PAF antagonist CV-6209 partially reversed the PAF induced effects on HTGL, TC and PL.

Animals↗
PubMed1990

Mitogenic signalling pathway of tumour necrosis factor involves the rapid tyrosine phosphorylation of 41,000-Mr and 43,000-Mr cytosol proteins.

Tumour necrosis factor (TNF) is a potent mitogen for some fibroblast cell lines. Here we have examined the TNF-mediated changes in protein phosphorylation in Swiss 3T3 and human FS-4 fibroblasts, and compared them with changes observed after the treatment of cells with other mitogens, such as platelet-derived growth factor (PDGF) and bombesin. TNF stimulated the rapid phosphorylation of two 41,000-Mr and two 43,000-Mr cytosol proteins on tyrosine, threonine and/or serine, as did PDGF, epidermal growth factor and fibroblast growth factor; the increased levels of this mitogen-induced protein-tyrosine phosphorylation correlated well with the extent of mitogen-induced DNA synthesis as determined by the percentage of labelled nuclei. In contrast, bombesin, which is an even better mitogen for Swiss 3T3 cells than TNF, stimulated the tyrosine phosphorylation of 41,000-Mr and 43,000-Mr proteins only to a limited extent. On the other hand, bombesin and PDGF stimulated the rapid serine phosphorylation of an 80,000-Mr acidic protein, a major substrate for protein kinase C; increased phosphorylation of the 80,000-Mr protein was not observed at all when cells were stimulated with TNF. These results suggest significant differences among the mitogenic signalling pathways of TNF, PDGF and bombesin as regards the involvement of protein kinases; the mitogenic signalling pathway of TNF involves the activation of tyrosine kinase, but not of protein kinase C, whereas bombesin seems to transduce its mitogenic signal mainly through the activation of protein kinase C, and the activation of both kinases seems to be involved in the mitogenic signalling pathway of PDGF.

Animals↗
PubMed1990

Mitogenic signaling pathways of growth factors can be distinguished by the involvement of pertussis toxin-sensitive guanosine triphosphate-binding protein and of protein kinase C.

We have examined the possible involvements of pertussis toxin (PT)-sensitive guanosine triphosphate (GTP)-binding protein (Gp) and protein kinase C (PKC) in the mitogenic signaling pathways of various growth factors by the use of PT-pretreated and/or 12-O-tetradecanoyl phorbol-13-acetate (TPA)-pretreated mouse fibroblasts. Effects of PT pretreatment (inactivation of PT-sensitive Gp) and TPA pretreatment (depletion of PKC) on mitogen-induced DNA synthesis varied significantly and systematically in response to growth factors: mitogenic responses of cells to thrombin, bombesin, and bradykinin were almost completely abolished both in PT- and TPA-pretreated cells; responses to epidermal growth factor (EGF), platelet-derived growth factor (PDGF), and vanadate were reduced to approximately 50% both in PT- and TPA-pretreated cells compared with native cells; response to basic fibroblast growth factor (bFGF) was not affected in PT-pretreated cells but was inhibited to some extent in TPA-pretreated cells. Thus, growth factors examined have been classified into three groups with regard to the involvements of PT-sensitive Gp and PKC in their signal transduction pathways. Binding of each growth factor to its receptor was not affected significantly by pretreatment of cells with PT or TPA. Inhibitory effects of PT and TPA pretreatment on each mitogen-induced DNA synthesis were not additive, suggesting that the functions of PT-sensitive Gp and PKC lie on an identical signal transduction pathway. Although all three groups of mitogens activated PKC, signaling of each growth factor depends to a varying extent on the function of PKC. Our results indicate that a single peptide growth factor such as EGF, PDGF, or bFGF acts through multiple signaling pathways to induce cell proliferation.

Animals↗
PubMed1990

[Glomerular apolipoprotein B deposition in glomerular diseases].

In an attempt to elucidate the relationship between the progress of glomerular injury and abnormalities of lipid metabolism, we investigated glomerular deposition of apolipoprotein B (apo B) in renal biopsy specimens from 60 patients with glomerular diseases by indirect immunofluorescence using antihuman apo B-100 monoclonal antibody in comparison with clinical and histopathological findings. The patients were divided into 2 groups according to the intensity in staining of apo B in glomerulus (group A: negative or weakly positive; and group B: definitely positive). Staining of apo B in glomerulus was found in 37 patients (62%). The levels of serum total cholesterol, phospholipids, low density lipoprotein cholesterol and apo B in group B were significantly higher than in group A. The urinary protein excretion in group B was greater than that in group A. Group B was also shown to have a significantly decrease in renal function. Light microscopy revealed severe mesangial proliferation in patients with IgA nephropathy of group B. These findings suggested that glomerular apo B containing-lipoprotein deposition may play an important role in the progression of glomerular injury.

Adolescent↗
PubMed1990

[A case of long-term survival of a patient with complicated diffuse metastatic leptomeningeal carcinomatosis secondary to lung adenocarcinoma].

A case of long-term survival of a female patient with complicated diffuse metastatic leptomeningeal carcinomatosis (DMLC) secondary to lung cancer is reported. A 36-year-old woman, hospitalized with a chief complaint of headache and unproductive cough, was diagnosed as having primary lung adenocarcinoma (T4N1M1 oss) and was given systemic chemotherapy. Although progressive deterioration of her headache continued, repeated neurological examination, cerebrospinal fluid (CSF) examination, and cranial CT scans failed to show evidence of metastasis to the central nervous system, and the only finding suggesting CNS involvement was an elevated CEA level in CSF. Later in the course of her treatment, the patient suddenly lost her vision and subsequently consciousness due to acute increased intracranial pressure, and emergency ventricular drainage was performed for therapeutic and diagnostic purposes. Malignant cells were found in CSF obtained from a ventricular drainage and she was treated successfully by systemic and intrathecal chemotherapeutic agents. She was discharged after a ventriculoperitoneal shunt operation for hydrocephalus; a double-dome reservoir was used for continuous intrathecal administration of the anticancer drugs, and a shunt filter was located in the tube to prevent the dissemination of cancer cells. In addition to methotrexate and cytosine arabinoside, ACNU and interleukin-2 were administered intrathecally without serious adverse effects, but no apparent therapeutic effects were noted either. She survived over 2 years after DMLC was first diagnosed. At autopsy DMLC secondary to lung adenocarcinoma was confirmed, but no evidence of leukoencephalopathy due to aggressive intrathecal chemotherapy was found. Current therapy for patients with DMLC and its clinical problems are discussed in relation to our experience in this case.

Adenocarcinoma↗
PubMed1989

Subpopulations of T alpha cells in patients with IgA nephropathy: correlation between T alpha 4 cells and in vitro IgA production.

T cells play important roles in the regulation of the immune system and are divided into subpopulations by various kinds of markers on the membrane surface. T cells with Fc-receptors for IgA are termed T alpha cells, and the properties of this cell population have been revealed in recent years. T alpha cells are increased in patients with IgA nephropathy and possess IgA specific helper activity. T alpha cells consist of two subpopulations, T alpha cells with OKT4 antigen (T alpha 4 cells) and with OKT8 antigen (T alpha 8 cells). To investigate the immunological aberrations in patients with IgA nephropathy, we detected immunoglobulin produced by peripheral blood lymphocytes and enumerated the numbers of T alpha cells (including both T alpha 4 and T alpha 8 cells). The numbers of T alpha 4 cells (but not T alpha 8 cells) and in vitro IgA production were increased in patients with IgA nephropathy (IgA nephropathy, mean 1.9%. Control, mean 0.8%. P = 0.0075). In addition, the numbers of T alpha 4 cells and the amount of IgA in the supernatant of lymphocyte cultures were positively correlated in these patients (P = 0.025. r = 0.3836). From the results in the present study, it was suggested that T alpha 4 cells might be related to immunological aberrations, such as an increase in IgA seen in patients with IgA nephropathy.

Adult↗