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N Kubota

Publications and source records attributed to N Kubota.

At least 19 recordsLinked to original sources
PubMed2000

A simple preparation of half N-acetylated chitosan highly soluble in water and aqueous organic solvents.

A simple and improved method of preparing highly soluble chitosan (half N-acetylated chitosan) was developed using a series of chitosan samples of low molecular weights, and the solubility of the half N-acetylated chitosan in water and organic solvents was investigated in detail. To reduce the molecular weight, chitosan was treated with NaBO3 under the condition that chitosan was homogeneously dissolved in aqueous acetic acid. Weight-average molecular weights of the obtained chitosan samples were determined using a size-exclusion chromatography system equipped with a low-angle laser light-scattering photometer. Each chitosan sample was then N-acetylated with acetic anhydride under the condition that chitosan was homogeneously dissolved in aqueous acetic acid again. The water solubility of the half N-acetylated chitosan thus prepared increased with decreasing molecular weight. From 1H NMR spectroscopy, it was suggested that the sequence of N-acetylglucosamine and glucosamine residues was random. The solubility of the half N-acetylated chitosan of low molecular weight was rather high even in aqueous dimethylacetamide and dimethylsulfoxide.

Acetylation↗
PubMed2000

Involvement of nuclear receptor coactivator SRC-1 in estrogen-dependent cell growth of MCF-7 cells.

Steroid hormones regulate cell growth and function through the transcriptional control of target genes by their cognate nuclear receptors. These receptors bind to ligands and associate with transcriptional cofactors to stimulate transcription. SRC-1, one of the nuclear receptor coactivators, is known to interact with nuclear receptors and enhance transactivation function in a ligand-dependent manner. In this study, to assess the function of SRC-1 in cell growth regulated by nuclear receptor ligands, we established a stable transformant cell line overexpressing human SRC-1 and studied the action of 17beta-estradiol (E(2)) on cell growth as well as the expression of E(2)-responsive genes in MCF-7 cells. We found that SRC-1 overexpression potentiates cell growth stimulated by E(2) in accordance with enhancement of transcriptional activation of exogenous and endogenous E(2)-responsive genes. These findings clearly indicate the importance of nuclear receptor coactivators for the activities of steroid/lipophilic vitamins in cell growth and gene expression.

Adenocarcinoma↗
PubMed2000

Proteomic approach to the identification of cell membrane proteins.

The expression of plasma membrane proteins in human monocyte-derived U937 cells was examined by cell disruption and isolation of microsomal fractions. Two alternative procedures for cell disruption, Dounce homogenization and nitrogen cavitation, were compared. Cell homogenization and sequential centrifugation resulted in an approximately fivefold enrichment of plasma membrane proteins in the microsomal fraction. However, identification of 30 such apparently enriched proteins by two-dimensional (2-D) electrophoresis, proteolytic digestion, and mass spectrometry revealed that only eight were plasma membrane proteins, the remaining 22 being contaminants. In contrast, nitrogen cavitation followed by sequential centrifugation and solubilization of proteins with sodium dodecyl sulfate (SDS) and 3-[(3-cholamidopropyl)dimethylammonio]-1-propane-sulfonate (CHAPS) detergent yielded subcellular fractions, including microsomes, that showed little overlap in constituent proteins as revealed by 2-D electrophoresis. These results highlight the importance of obtaining pure plasma membranes and complete solubilization of membrane proteins for proteomic analysis.

Electrophoresis, Gel, Two-Dimensional↗
PubMed2000

Retrospective analysis of type of hepatic resection for hepatocellular carcinoma.

BACKGROUND: Hepatic resection for hepatocellular carcinoma is now considered a relatively safe procedure, owing to refined surgical techniques and management. METHODS: Three hundred different types of hepatic resection performed in one hospital were reviewed retrospectively. The operative morbidity and mortality rates and long-term survival were assessed. RESULTS: Tumours were extirpated by lobectomy or extended lobectomy in 48 patients, by segmentectomy in 53, by subsegmentectomy in 94 and by partial resection in 105. The overall hospital mortality rate was 4 per cent (13 of 300). No operative death occurred after lobectomy or extended lobectomy. Segmentectomy and subsegmentectomy were associated with high postoperative complication and hospital mortality rates. There was a significant difference in survival rate between patients with and without a cancer-free surgical margin of more than 1.0 cm. The overall 5- and 9-year survival rates were 51 and 9 per cent respectively. Rates were 56 and 24 per cent for lobectomy or extended lobectomy. CONCLUSION: For tumours of 2.1 cm or more in size lobectomy appears to carry the least risk of postoperative complications and death in hospital and best achieves a cancer-free surgical margin.

Carcinoma, Hepatocellular↗
PubMed2000

Pancreaticojejunostomy versus pancreaticogastrostomy in reconstruction following pancreaticoduodenectomy.

BACKGROUND: Pancreaticogastrostomy (PG) after pancreaticoduodenectomy has been reported to have a lower incidence of pancreatic fistula than pancreaticojejunostomy (PJ) but this was not confirmed in a recent prospective randomized study. METHODS: Different methods of reconstruction after pancreaticoduodenectomy were used between January 1994 and January 1999 in two university-affiliated hospitals, PJ (n = 69) in one hospital and PG (n = 73) in the other. Operations at both hospitals were performed by the same surgical team. All pancreatic anastomoses were carried out in two layers with pancreatic duct stents. Pancreatic fistula was identified by the presence of more than 1000 units/l of amylase-rich fluid in the drains 7 days or more after operation, by radiography from the pancreatic duct stent and by water-soluble contrast upper gastrointestinal studies. RESULTS: The two groups of patients were similar in terms of age, sex, findings at preoperative assessment, disease status, operative time, intraoperative blood loss and nature of non-tumorous pancreatic tissue. The amylase level in ascites at 7 days after operation was significantly lower after PG than PJ (P = 0.032). The incidence of pancreatic fistula in the PG group (zero) was significantly less than that after PJ (13 per cent) (P = 0.014). Intra-abdominal haemorrhage and intra-abdominal abscess occurred in three (4 per cent) and four patients (6 per cent) respectively, with two hospital deaths (3 per cent) in the PJ group, but these complications did not occur after PG. CONCLUSION: This controlled clinical study supports the hypothesis that PG is safer than PJ, particularly with regard to the incidence of pancreatic fistula.

Aged↗
PubMed2000

Multimodality treatment for patients with hepatocellular carcinoma: a single institution retrospective series.

BACKGROUND: The main therapeutic options for hepatocellular carcinoma (HCC) are hepatic resection, transcatheter arterial embolization (TAE), percutaneous ethanol injection therapy (PEIT) and regional chemotherapy (RC). METHODS: This study retrospectively examined the results of primary treatment of 600 patients with hepatocellular carcinoma selected according to the treatment guidelines of our facility and the results of various combination therapies for recurrent cases. The selection criteria of therapeutic options included the number and size of tumours and hepatic function. RESULTS: The selected primary treatment was hepatic resection for 53.7% of the cases, TAE for 31.5%, PEIT for 8.2% and RC for 6.6%,. The treatment for post-resection recurrence was TAE alone for 62.4% of the cases, TAE + RC for 4.0%, PEIT for 15.2%, TAE + PEIT alone for 4.8%, RC for 8.0% and hepatic resection for 5.6%. The treatment for post-TAE recurrence was TAE alone for 83% of the cases, TAE + PEIT for 9%, TAE + RC for 3%, RC alone for 3% and PEIT alone for 2%. For post-PEIT, therapy was PEIT alone for 71.4% of the cases and PEIT + TAE for 28.6%. For post-RC, RC alone was used for 92.5% and RC + PEIT for 7.5%. The cumulative 3 and 5-year survival rates were 84.4% and 70.6%, respectively for stage I; 61.5% and 48.6% for stage II; 52.7% and 20.5% for stage III; and 22.8% and 17.1% for stage IVA. The cumulative 5 and 7-year survival rates after the primary treatments were 52.% and 40.1%, respectively, for hepatic resection; 46.5% and 38.7%, for TAE; 49.6% and 33.1% for PEIT; and 16.7% and 8.3% for RC. CONCLUSIONS: To improve the treatment results for HCC, early detection is essential and various modalities of treatments in combination should be used for recurrence after primary treatment.

Adult↗
PubMed2000

Effect of hemostatics used during operations for digestive organ on cancer cells present in the peritoneal cavity.

We investigated effects of hemostatics used during operations for digestive organ on cancer cells present in the peritoneal cavity using BALB/c mice inoculated with Meth A tumor cells (fibrosarcoma) intraperitoneally (i.p.) and C3H/He mice inoculated with MH134 tumor cell (hepatic cell carcinoma) i.p. Microfibrillar collagen hemostat (Avitene) or fibrinogen preparation (Beriplast P) did not affect survivals of those tumor-bearing mice. Gelatin sponge (Spongel)prolonged survivals of MH134 tumor-bearing mice. Liquid form gelatin used instead of Spongel displayed in vitro antitumor effect on MH134 tumor cells at the concentration of 15 mg/ml. Radioactive sodium chromate-labeled MH134 and Meth A tumor cells were not lysed when they were incubated with 15 mg/ml of liquid form gelatin for 24 hours. On the other hand, the tritium thymidine (3H-TdR) uptake by MH134 or RL male 1 tumor cells was suppressed when they were incubated with 15 mg/ml of liquid form gelatin for 24 hours. Proliferation of Meth A tumor cells were not affected by the treatment. Effect of liquid form gelatin on phytohemagglutinin (PHA)-stimulated spleen cells as a benign counter-part of RL male 1 tumor cells (T cell lymphoma) was examined. Liquid form gelatin (15 mg/ml) did not suppress 3H-TdR uptake by PHA-stimulated spleen cells.

Animals↗
PubMed2000

Insulin receptor substrate-1 in osteoblast is indispensable for maintaining bone turnover.

Insulin receptor substrates (IRS-1 and -2) are essential for intracellular signaling by insulin and IGF-I, anabolic regulators of bone metabolism. Mice lacking the IRS-1 gene IRS-1(-/-) showed severe osteopenia with low bone turnover. IRS-1 was expressed in osteoblasts, but not in osteoclasts, of wild-type (WT) mice. IRS-1(-/-) osteoblasts treated with insulin or IGF-I failed to induce tyrosine phosphorylation of cellular proteins, and they showed reduced proliferation and differentiation. Osteoclastogenesis in the coculture of hemopoietic cells and osteoblasts depended on IRS-1 expression in osteoblasts and could not be rescued by IRS-1 expression in hemopoietic cells in the presence of not only IGF-I but also 1,25(OH)(2)D(3). In addition, osteoclast differentiation factor (RANKL/ODF) was not induced by these factors in IRS-1(-/-) osteoblasts. We conclude that IRS-1 deficiency in osteoblasts impairs osteoblast proliferation, differentiation, and support of osteoclastogenesis, resulting in low-turnover osteopenia. Osteoblastic IRS-1 is essential for maintaining bone turnover, because it mediates signaling by IGF-I and insulin and, we propose, also by other factors, such as 1,25(OH)(2)D(3).

Animals↗
PubMed2000

Insulin effect during embryogenesis determines fetal growth: a possible molecular link between birth weight and susceptibility to type 2 diabetes.

Low birth weight has been reported to be associated with impaired insulin secretion and insulin resistance. It has been proposed that this association results from fetal programming in response to the intrauterine environment (the thrifty phenotype hypothesis). To elucidate the relationship between birth weight and genetically determined defects in insulin secretion, we measured the birth weights of neonates derived from crosses of male pancreatic beta-cell type glucokinase knockout (Gck+/-) mice and female wild-type (WT) or Gck+/- mice. In 135 offspring, birth weights were lower in the presence of a fetal heterozygous mutation and higher in the presence of a maternal heterozygous mutation. Moreover, Gck-/- neonates had significantly smaller birth weights than WT or Gck+/- neonates (means +/- SE 1.49+/-0.03 [n = 30] vs. 1.63+/-0.03 [n = 30] or 1.63+/-0.02 [n = 50] g, respectively; P<0.01). Thus, Gck mutations in beta-cells may impair insulin response to glucose and alter intrauterine growth as well as glucose metabolism after birth. This study has confirmed the results of a previous report that human subjects carrying mutations in Gck had reduced birth weights and has provided direct evidence for a link between insulin and fetal growth. Moreover, birth weights were reduced in insulin receptor substrate-1 knockout mice despite normal insulin levels. Taken together, these results suggest that a genetically programmed insulin effect during embryogenesis determines fetal growth and provides a possible molecular link between birth weight and susceptibility to type 2 diabetes.

Animals↗
PubMed1999

Beta(2)-microglobulin identified as an apoptosis-inducing factor and its characterization.

Major histocompatibility complex (MHC) molecules play an important role in antigen presentation for induction of tumor as well as cellular and humoral immunities. Recent studies using anti-MHC antibodies demonstrated that antibodies specific for HLA class I molecules induced cellular activation and a type of apoptosis that may be distinct from Fas-dependent or TNFR (tumor necrosis factor-alpha receptor)-dependent processes. We purified a previously untested apoptosis-inducing factor from HL-60 human leukemic cell-conditioned media to homogeneity and sequenced it. It was identified as beta(2)-microglobulin (beta(2)m), which has been previously known as thymotaxin and is a part of the HLA class I antigen complex. beta(2)m acts on both T-leukemic cells and myeloid leukemic cells to induce apoptosis, which then activates caspase 1 and 3. Cross-linking studies showed that biotinilated beta(2)m recognized an epitope distinct from those recognized by the anti-HLA class I antibody, as reported previously. We demonstrated that beta(2)m plays a previously unrecognized and important role in regulating the elimination of tumor cells, which occurs as a result of the action of beta(2)m as an apoptosis-inducing factor.

Amino Acid Chloromethyl Ketones↗
PubMed1999

Role of NADH shuttle system in glucose-induced activation of mitochondrial metabolism and insulin secretion.

Glucose metabolism in glycolysis and in mitochondria is pivotal to glucose-induced insulin secretion from pancreatic beta cells. One or more factors derived from glycolysis other than pyruvate appear to be required for the generation of mitochondrial signals that lead to insulin secretion. The electrons of the glycolysis-derived reduced form of nicotinamide adenine dinucleotide (NADH) are transferred to mitochondria through the NADH shuttle system. By abolishing the NADH shuttle function, glucose-induced increases in NADH autofluorescence, mitochondrial membrane potential, and adenosine triphosphate content were reduced and glucose-induced insulin secretion was abrogated. The NADH shuttle evidently couples glycolysis with activation of mitochondrial energy metabolism to trigger insulin secretion.

Adenosine Triphosphate↗
PubMed1999

Proposed standard for human blood vitamin B1 value using HPLC. The Committee for Vitamin Laboratory Standards, Japan.

Standard reference ranges for all laboratory test values are mandatory. This study was designed to establish a reference range for blood vitamin B1 levels, since the normal range has not been determined in the Japanese population. We founded the Japan Committee for Vitamin Laboratory Standards, which was incorporated with the Vitamin Society of Japan and the Japanese Society of Nutrition and Food Science. We standardized whole blood vitamin B1 levels using three HPLC techniques (post-column reverse-phase HPLC, pre-column reverse-phase HPLC, and precolumn GP-HPLC). The reference range was obtained in 54 volunteers administered a 1,800 kcal diet with 2 mg of vitamin B1 (1.74 mg measured) daily to avoid marginal vitamin B1 deficiency in the population. The range for each assay was 26-47, 28-51, and 28-56 ng/ml, respectively. Our data suggest that 26-28 ng/ml is the lower limit of normal for whole blood vitamin B1, but further studies in a larger population are needed in order to obtain more definitive results.

Chromatography, High Pressure Liquid↗
PubMed1999

Pharmacological characterization of dopamine transport in cultured rat astrocytes.

The effects of GBR-12909 (selective DA uptake inhibitor), zimelidine (selective 5-HT uptake inhibitor) and nisoxetine (selective NE uptake inhibitor) on the uptake of 30 nM [3H]DA into cultured rat astrocytes were examined. [3H]DA uptake was inhibited by approximately 50% by GBR-12909 or zimelidine in a concentration-dependent manner (100 nM to approximately 10 microM). Furthermore, the inhibition curves of GBR-12909 were biphasic, and uptake was completely inhibited by a high concentration of GBR-12909 (100 microM). [3H]DA uptake was also inhibited by approximately 50% by nisoxetine in a concentration-dependent manner (0.1 to approximately 100 nM), and nisoxetine was more potent than GBR-12909 or zimelidine. The inhibitory potencies were in the order nisoxetine > GBR-12909 > zimelidine. The uptake of [3H]DA under Na+-free conditions was approximately 50% of that under normal conditions. Thus, DA was taken up by both Na+-dependent and Na+-independent mechanisms. Nisoxetine (100 nM), zimelidine (100 microM) and GBR-12909 (10 microM) inhibited [3H]DA uptake into astrocytes only in the presence of Na+. On the other hand, this uptake was completely inhibited by a high concentration of GBR-12909 (100 microM) in the absence of Na+. The present data suggest that the Na+-dependent uptake of [3H]DA in cultured rat astrocytes may occur in the NE uptake system. Furthermore, astrocytes express the extraneuronal monoamine transporter (uptake2), which is an Na+-independent system, and this transporter is involved in the inactivation of centrally released DA.

Animals↗
PubMed1999

Regulation of dopamine uptake by basic fibroblast growth factor and epidermal growth factor in cultured rat astrocytes.

We examined the characteristics of dopamine (DA) uptake and its regulation by neurotrophic factors such as basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) in cultured rat astrocytes. In the presence of inhibitors of monoamine oxidase (MAO) and catechol-O-methyl-transferase (COMT), astrocytes took up DA by Na(+)-dependent and Na(+)-independent mechanisms that were sensitive to a reduction in temperature. The Na(+)-dependent and Na(+)-independent components increased linearly with increasing [3H]DA concentration (1-1000 microM), and showed no saturation. Na(+)-dependent DA uptake was significantly inhibited by ouabain, a Na(+)-K+ ATPase inhibitor. In bFGF-treated astrocytes, [3H]DA uptake increased in a time-dependent manner until 48 h, and declined after 72 h in both the presence and absence of Na+. In EGF-treated astrocytes, [3H]DA uptake increased in a time-dependent manner until 72 h in both the presence and absence of Na +. This enhancement of DA uptake induced by EGF or bFGF was significantly inhibited when the cells were cultured with actinomycin D, cycloheximide, or brefeldin A. Actinomycin D and brefeldin A also significantly inhibited the basal uptake of [3H]DA into astrocytes. These results suggest the existence of Na(+)-dependent and Na(+)-independent DA uptake in cultured rat astrocytes, and that EGF or bFGF might stimulate the expression and translocation of the extraneuronal DA transporter.

Animals↗
PubMed1999

PPAR gamma mediates high-fat diet-induced adipocyte hypertrophy and insulin resistance.

Agonist-induced activation of peroxisome proliferator-activated receptor gamma (PPAR gamma) is known to cause adipocyte differentiation and insulin sensitivity. The biological role of PPAR gamma was investigated by gene targeting. Homozygous PPAR gamma-deficient embryos died at 10.5-11.5 dpc due to placental dysfunction. Quite unexpectedly, heterozygous PPAR gamma-deficient mice were protected from the development of insulin resistance due to adipocyte hypertrophy under a high-fat diet. These phenotypes were abrogated by PPAR gamma agonist treatment. Heterozygous PPAR gamma-deficient mice showed overexpression and hypersecretion of leptin despite the smaller size of adipocytes and decreased fat mass, which may explain these phenotypes at least in part. This study reveals a hitherto unpredicted role for PPAR gamma in high-fat diet-induced obesity due to adipocyte hypertrophy and insulin resistance, which requires both alleles of PPAR gamma.

Adipocytes↗
PubMed1999

Mutation induction and RBE of low energy neutrons in V79 cells.

We have examined the neutron energy dependency of cell killing and mutation induction at the hprt locus in Chinese hamster V79 cells. Monoenergetic neutrons at 0.32, 0.57, and 1.2 MeV were generated at the Hiroshima University Radiobiological Research Accelerator (HIRRAC) Facility, and were used to irradiate cells. The variation in RBE with neutron energy for the end points of cell survival and hprt mutation induction was observed. When compared to 137Cs gamma-rays, all neutron energies were more effective at both cell killing and induction of mutation. Over the range of the neutron energies examined, we found that cytotoxicity increased as the energy decreased from 1.2 to 0.32 MeV. In comparison to gamma-rays, RBEs for cell lethality at 10% survival were 5.7, 6.7, and 7.6 for 1.2, 0.57, and 0.32 MeV, respectively. Mutation induction, on the other hand, was highest at 0.57 MeV with a gradual decrease at 1.2 and 0.32 MeV. RBEs for mutation induction were 9.7, 19.4, and 13.9 for 1.2, 0.57, and 0.32 MeV neutrons. We isolated independent V79 cell mutants at the hprt locus from untreated and neutron-exposed cells and determined the genetic changes underlying the mutation by multiplex polymerase chain reaction (PCR)-based exon deletion analysis. Preliminary results are suggestive of a specific relationship between deletion pattern and neutron energy.

Animals↗
PubMed1999

Computerized tomography nephroscopic images of renal pelvic carcinoma.

PURPOSE: Computerized tomography (CT) endoscopy is an interactive 3-dimensional image acquired by helical CT. We assess the usefulness of CT nephroscopy in the diagnosis of renal pelvic cancers. MATERIALS AND METHODS: Surface rendering CT nephroscopy was performed after intravenous administration of contrast agent and furosemide in 32 patients with suspected renal pelvic carcinoma. Retrospective review revealed that 23 patients later underwent nephroureterectomy. Two observers blinded to the pathology results independently reviewed the CT nephroscopic and axial CT images of the 23 patients to localize pelvic tumors and to determine the extent of the disease. The CT nephroscopic and axial CT images were correlated with the pathological findings. RESULTS: Pathological examination revealed 4 polypoid, 6 pedunculated, 7 sessile, 1 sessile with ulceration and 6 infiltrating renal pelvic carcinomas in the 23 patients. CT nephroscopy revealed 22 carcinomas (92%) and CT demonstrated 20 (83%). CT nephroscopy was superior to axial CT in detecting 4 pedunculated and 3 infiltrating carcinomas but failed to detect 1 sessile carcinoma which completely replaced the upper portion of a double renal pelvis and 1 polypoid tumor associated with infiltrating carcinoma. The CT nephroscopic images correlated well with pathological findings. CONCLUSIONS: CT nephroscopy is useful to visualize the complex morphology of renal pelvic carcinomas noninvasively and is superior to axial CT for the detection of infiltrating and pedunculated carcinomas.

Adult↗