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Hiroshi Inoue

Publications and source records attributed to Hiroshi Inoue.

At least 217 records · Page 12Linked to original sources
PubMed2002

G protein gamma 7 expression as a new clinicopathological marker in patients with intrahepatic cholangiocarcinoma.

HYPOTHESIS: The signal alterations mediated by small G proteins such as Ras, Rho, and Rac have been reported in several cancers. The human G protein gamma 7 (G-gamma 7) gene, which is down-regulated in various digestive organ cancers, was recently identified and cloned. Thus, the G-gamma 7-coupled heterotrimeric G proteins may also contribute to carcinogenesis in human cancers. SETTING: University hospital and medical institute of bioregulation. PATIENTS AND METHODS: The clinicopathological significance of G-gamma 7 expression in 18 patients with intrahepatic cholangiocarcinoma (IHCC) was examined. The tumor-nontumor ratio of G-gamma 7 expression was determined using reverse transcription polymerase chain reaction analysis. To visualize the localization of G-gamma 7, an immunohistochemical study was performed. MAIN OUTCOME MEASURE: Clinicopathological significance of G-gamma 7 expression in human IHCC. RESULTS: Expression of G-gamma 7 messenger RNA was lower in tumor tissue than in the corresponding nontumor tissue in 17 (94%) of 18 patients with IHCC. The mean tumor-nontumor ratio was 0.54. Eleven patients with tumor-nontumor ratios less than 0.5 showed significantly poorer differentiated IHCC than 7 with tumor-nontumor ratios of 0.5 and greater (P<.01). Decreased expression of G-gamma 7 protein in the carcinoma tissue, especially in the poorly differentiated IHCC tissue, was confirmed using immunohistochemical staining. CONCLUSIONS: Reduced expression of G-gamma 7 is associated with the histological grade of IHCC and may therefore prove to be a useful marker for predicting the biological aggressiveness of human IHCC.

Adult↗
PubMed2002

Lipid rafts as the signaling scaffold for NK cell activation: tyrosine phosphorylation and association of LAT with phosphatidylinositol 3-kinase and phospholipase C-gamma following CD2 stimulation.

Natural killer (NK) cells participate in both innate and adaptive immunity through the prompt secretion of cytokines and ability to lyse virally infected cells or tumor cells. Although it has been well understood that lipid rafts (rafts) and a raft-associated linker for activation of T cells (LAT) plays a central role in TCR signal transduction, there are still great gaps in our knowledge of the molecular events involved in NK cell activation. We show here that CD2 and rafts became polarized to the site of NK cell activation by CD2 cross-linking or target cell binding using confocal microscopy, and LAT and a significant amount of CD2 colocalized in raft fractions of sucrose-density gradient from an NK cell line, NK3.3. CD2 cross-linking strongly induced tyrosine phosphorylation of LAT, resulting in increased association with phosphatidylinositol 3-kinase (PI 3-K) and phospholipase C-gamma1 (PLC-gamma1). In vitro binding studies using glutathione S-transferase fusion proteins demonstrated that a large portion of the association between LAT and PI 3-K or PLC-gamma1 was mediated through their SH2 domains in tyrosine phosphorylation-dependent manner. Furthermore, disruption of lipid rafts by cholesterol depletion from cell membranes using methyl-beta-cyclodextrin markedly reduced LAT tyrosine phosphorylation and NK cell functions, including cytotoxicity and granule exocytosis. These results document that modulation of raft integrity by aggregation of NK cell activating receptors, which leads to the formation of complexes of LAT with PI 3-K and PLC-gamma1, is essential for the NK cell lytic mechanisms.

Adaptor Proteins, Signal Transducing↗
PubMed2002

A single-nucleotide polymorphism of SMARCB1 in human breast cancers.

The gene SMARCB1 has been considered a candidate for a tumor-suppressor gene. Nucleotide alterations in SMARCB1 have been reported, primarily in association with malignant rhabdoid tumor cases. We carried out a search for mutations in SMARCB1 in 60 human gastro-intestinal tract carcinoma cases, 122 breast cancer cases, and 36 human cancer cell lines. A single-nucleotide polymorphism (SNP) at codon 152 with an amino acid change (Asn to Asp) was found in 2 of 122 (1.6%) breast cancer cases, and another SNP at codon 299 without an amino acid change was found in tumor and normal tissues from 7 (5.7%) cases. Codons 152 and 299 of SMARCB1 are localized near or within the binding site for the cMYC protein. The amount of immunoprecipitated cMYC protein was reduced in two different cell lines expressing the codon 152 polymorphic SMARCB1 clone compared with those expressing wild-type SMARCB1, regardless of the identical expression of SMARCB1 protein in both cell lines. Therefore, the SNP at codon 152 is considered to be one of the coding SNPs that alters the SMARCB1-cMYC complex, which regulates various tumor-suppressor related genes against cancer. In addition, we identified three types of splicing isoforms, a 27-bp deleted gene, a 51-bp inserted gene, and a consensus gene, in both carcinoma tissues and in normal tissues; however, no clinical significance was observed for those isoforms. We found a nucleotide change at codon 152 of SMARCB1 that may alter the amount of immunoprecipitated cMYC protein, but we finally determined that SMARCB1 is highly conserved in human solid carcinomas.

Base Sequence↗
PubMed2002

The evaluation of dysphagia following radical surgery for oral and pharyngeal carcinomas by cine-magnetic resonance imaging (Cine-MRI).

Cine-magnetic resonance imaging (cine-MRI) creates moving pictures by a video system and turbo-flash method that allow for high-speed MRI. This report describes our experience using this new technique for dynamic imaging using the fast spoiled GRASS (SPGR) sequence to study swallowing in patients with dysphagia following radical surgery for oral cancer. We defined two new parameters, laryngeal elevation and the angle of the epiglottis, to quantify swallowing ability by cine-MRI. These variables were markedly different in patients with dysphagia than they were in healthy controls. Cine-MRI not only provides dynamic images of swallowing but can generate objective measures of swallowing ability as well.

Adult↗
PubMed2002

Effects of long-term renal sympathetic denervation on heart failure after myocardial infarction in rats.

The purpose of the present study was to investigate the effects of long-term renal denervation (RD) on heart failure due to myocardial infarction (MI). Wistar rats were anesthetized and the bilateral renal nerves were surgically denervated 2 days before MI was induced by coronary artery ligation. Four weeks later, left ventricular (LV) function and sodium excretion were determined. In MI rats, RD improved the reduced sodium excretion. MI + RD rats revealed lower LV end-diastolic pressure and greater maximum dP/dt as compared with those of MI+ innervation (INN) rats. LV end-diastolic and end-systolic dimensions were significantly smaller and LV fractional shortening was greater in MI + RD rats than in MI + INN rats (20.9% +/- 3.2% vs 14.9% +/- 3.0%). In rats without MI, RD did not affect either sodium excretion or LV function and dimensions. The present results suggest that the long-term RD reduces LV filling pressure and improves LV function after MI, probably due to a restoration of impaired natriuresis. Increased renal sympathetic nerve activity might contribute to the progression of heart failure after MI.

Animals↗
PubMed2002

Role for adapter proteins in costimulatory signals of CD2 and IL-2 on NK cell activation.

Natural killer (NK) cells participate in both innate and adaptive immunity through the prompt secretion of cytokines and ability to lyse virally infected cells or tumor cells. Triggering of NK cells requires aggregation of surface receptors such as CD2 and CD16, and NK cell activity can be augmented in vitro by stimulation with IL-2. In this study, we examined the role of adapter proteins in the increased NK activation following CD2 crosslinking and IL-2 stimulation of NK3.3 cells. NK3.3 cells lysed NK-sensitive K562 cells in a CD2-dependent manner, and IL-2 markedly enhanced lytic activity in a 4h cytotoxic assay. IL-2 also enhanced spontaneous and CD2-mediated granule exocytosis from NK3.3 cells. CD2 crosslinking markedly induced tyrosine phosphorylation of Cbl associated with Grb2 or CrkL, Shc and LAT, compared with IL-2 stimulation. However, costimulation of IL-2 with CD2 crosslinking remarkably enhanced associations of Grb2-Shc and CrkL-Cbl, compared to IL-2 stimulation or CD2 crosslinking alone. In vitro binding studies using GST-fusion proteins revealed that interactions of Grb2-Shc and CrkL-Cbl were mediated through each SH2 domain in tyrosine phosphorylation-dependent manner. Furthermore, CD2 crosslinking, but not IL-2 stimulation, markedly induced tyrosine phosphorylation of LAT. Thus, tyrosine phosphorylation of different adapter proteins and consequent interactions between signaling molecules described here may explain the molecular mechanisms of the additive effects of IL-2 stimulation and CD2 crosslinking on NK cell activation.

Adaptor Proteins, Signal Transducing↗
PubMed2002

Disparate force-frequency effects of pimobendan and dobutamine in conscious dogs with tachycardia-induced cardiomyopathy.

BACKGROUND: This study was designed to examine how a calcium sensitizer, pimobendan, affected a force-frequency response (FFR) as compared to the beta-adrenergic agonist dobutamine. METHODS AND RESULTS: Left ventricular (LV) contractility and relaxation were evaluated by the slope (Ees) of the LV end-systolic pressure-volume relation and the time constant (Tau) of LV pressure decay. Using 6 conscious dogs with tachycardia-induced heart failure, the FFR was examined before and after administration of dobutamine (6 microg/kg/min) or pimobendan (0.5 mg/kg). Despite the similar inotropic and lusitropic action at the baseline heart rate, pimobendan and dobutamine showed different FFR and relaxation-frequency responses. Before administration of these drugs, there was no significant increase in LV contractility and relaxation by increasing heart rate. However, dobutamine amplified FFR (Ees: +3.1 +/- 1.4, P <.05) as compared with Ees for a comparable increase in heart rate before administration of the drug. On the other hand, pimobendan showed relatively mild amplification of FFR compared with dobutamine (Ees: +1.9 +/- 1.1, P <.05). The relaxation-frequency response tended to increase with dobutamine but not with pimobendan. CONCLUSIONS: Mild amplification of FFR observed in pimobendan suggests that this agent could be used more safely than beta-adrenergic agent when heart rate is increased, as seen with exercise.

Adrenergic beta-Agonists↗
PubMed2002

Age-related changes in the immunoreactivity of the monocyte/macrophage system in rat molar pulp after cavity preparation.

OBJECTIVE: Our objective was to compare the response of the monocyte/macrophage system of dental pulp to cavity preparation in aged rats (12 and 18 months old) with that seen in young adult rats (3 and 6 months old). STUDY DESIGN: Cavities were prepared on the upper first molars, and the lower first molars served as intact controls. Specimens were collected at 1 day after cavity preparation, and cryostat sections were made. RESULTS: Accumulation of OX6+ antigen-presenting cells along the pulp-dentin border and a marked increase in cell size ED2+ resident macrophages were noted in both young adult and aged rats after cavity preparation. In both cases, the number of ED1+ cells increased significantly after cavity preparation because of infiltrating monocytes. CONCLUSION: These findings suggest that the pulpal defense reaction of the monocyte/macrophage system to cavity preparation in aged rats does not differ markedly from that in young adult rats.

Age Factors↗
PubMed2002

Analysis of the gene-expression profile regarding the progression of human gastric carcinoma.

BACKGROUND: Tumor tissue consists of a variable mixture of tumor and host-cell populations. Recent developments in laser microdissection (LMD) and cDNA microarray analysis have encouraged us to study the differential gene expression profiles among normal cells, primary carcinoma cells, and metastatic carcinoma cells in cases of gastric carcinoma. METHODS: Total RNA was extracted from the cells obtained by means of LMD from the primary carcinoma, the corresponding gastric epithelium, and the lymph node metastasis in 5 cases of primary gastric carcinoma. RNA was amplified by the T7-based amplification system to be applied to a cDNA microarray. Thereafter, the differentially expressed genes among the 3 populations were evaluated. RESULTS: cDNA samples for microarray studies were successfully obtained from each cell population of 5 cases. The cDNA microarray demonstrated that several interesting genes, such as cell-cycle regulators and growth factors, were overexpressed in the metastatic cells compared with in the primary carcinoma cells. Oncogenes and cell-adhesion molecules were more overexpressed in the primary carcinoma cells than in the normal cells. On the other hand, caspase 8 and cadherin were more suppressed in the primary carcinoma cells than in the normal cells. Interestingly, among the matrix metalloproteinase family, only MMP7 was identified as a differentially overexpressed gene in both the primary carcinoma and the metastatic cells in comparison with the normal cells. CONCLUSIONS: This study demonstrated that the combined use of LMD, T7-based amplification, and a cDNA microarray enabled us to identify genes directly associated with each population of tumor tissue. The method will open up new possibilities for the precise gene analysis of tumor progression and metastasis.

Adenocarcinoma↗
PubMed2002

Effects of leukocytosis and macrophage activation on anti-Thy 1.1 glomerulonephritis in the rat.

OBJECTIVE: In anti-Thy 1.1 proliferative glomerulonephritis (GN), glomerular infiltration of polymorphonuclear leukocytes (PMNs) reaches a peak level after 1 h and that of macrophages does so 24 h after induction. However, the roles of PMNs and macrophages in the pathogenesis of anti-Thy 1.1 GN remain unclear. We examined the effects on this model of leukocytosis induced by granulocyte colony-stimulating factor (G-CSF) and of macrophages stimulated by macrophage colony-stimulating factor (M-CSF). MATERIAL AND METHODS: Anti-Thy 1.1 GN was induced in male Wistar rats by intravenous injection of OX-7, a monoclonal antibody to the Thy 1 antigen. G-CSF (10 microg/kg/day), M-CSF (20 microg/kg/day) or vehicle was administered intraperitoneally for 7 days starting 24 h before the injection of OX-7. Histological examination of renal biopsy specimens was performed on Days 1, 5 and 14 after induction. RESULTS: Circulating and glomerular-infiltrating PMNs (RP-3-positive cells) were increased at Day 5 in G-CSF-treated rats compared with controls receiving vehicle, and glomerular mesangiolysis continued at Day 5. The number of proliferating cells positive for proliferating cell nuclear antigen at Day 5 and matrix scores at Day 14 were smaller in G-CSF-treated rats than in control rats. The mesangiolysis score was significantly higher in the G-CSF group than in the control group at Days 5 and 14, but not at Day 1. ED-1-positive cells were increased in number at Day 5 and matrix accumulation decreased at Day 14 in M-CSF-treated rats compared with controls. Serum creatinine level at Day 14 was lower in the M-CSF group, but not in the G-CSF group, compared with the control group. CONCLUSIONS: Activated macrophages may inhibit excess matrix accumulation and ameliorate the recovery of renal function, whereas leukocytosis inhibits the repair of mesangial cell injury in this model.

Animals↗
PubMed2002

Regional expression of CXCL12/CXCR4 in liver and hepatocellular carcinoma and cell-cycle variation during in vitro differentiation.

The CXCL12 / CXCR4 system may be important in carcinoma. Expression of the alpha-chemokine SDF-1alpha (stromal cell derived factor-1alpha) / CXCL12 mRNA is reduced in many carcinomas, yet its tissue protein expression may guide metastasis. Here we first compare the mRNA and protein expression of CXCL12 and its receptor CXCR4 in human liver, hepatocellular carcinoma, and malignant cell lines, and then assess cell cycle variation in CXCR4 expression. CXCR4 mRNA was present in most normal human tissues and malignant cell lines; it was only marginally reduced in hepatomas, while CXCL12 was markedly reduced, P < 0.0001. Immuno-histochemical staining of adjacent non-malignant liver showed regional CXCR4 cytoplasmic and cell-surface staining, limited to those hepatocytes around the central vein, a distribution resembling that of CXCL12. CXCL12 protein was not present in hepatocellular carcinoma cells in vivo, nor was cytoplasmic CXCR4 staining; nuclear CXCR4 protein expression in some malignant hepatocytes and CXCR4 staining of capillary endothelial cells around tumor cells were noted. In some malignant cell lines that had no CXCL12 on northern blots CXCL12 was weakly detectable by RT-PCR or protein staining in the cytoplasm of a few cells. With a view to future manipulation of CXCL12 / CXCR4 expression and growth we noted that in HT-29 cells CXCR4 protein expression was less on confluent than on non-confluent cells and varied during the cell cycle. Higher expression was associated most closely with the percentage of cells in the S-phase and inversely with the percentage of cells in the G1-phase. Treatment of HT-29 cells with butyrate reduced CXCR4 cell surface expression and reduced the percentage of cells in S-phase. In summary, CXCL12 protein expression parallels its mRNA, being markedly reduced in malignant cell lines and hepatomas; in liver, the regional distributions of CXCL12 and cytoplasmic CXCR4 are similar; finally, in HT-29, CXCR4 expression correlates with the S-phase of the cell cycle and is reduced during butyrate-induced differentiation.

Blotting, Northern↗
PubMed2002

Sequence variants in the FcepsilonRI alpha chain gene.

There is a relationship between IgE levels and expression of high-affinity IgE receptors (FcepsilonRI). Because the alpha chain is the only portion of the receptor that binds directly to IgE, we reasoned that sequence variants in the FcepsilonRI alpha gene may exist that alter these binding events. We screened all of the exons and the promoter region of the FcepsilonRI alpha chain gene with genomic DNA from 389 asthmatic and 341 normal control subjects for mutations by using single-stranded conformational polymorphism analysis. No nonsynonomous single nucleotide polymorphisms (SNPs) were identified in the coding region. Three SNPs were found in the promoter region: an A/C transversion at -770 from the translation start site; a G/A transition at -664; and a T/C transition at -335. No differences in allele frequencies were detected between asthmatic subjects and controls. Homozygosity for the C variant at locus -335 was more common in Caucasian asthmatic patients with IgE levels in the lower quartile than in the upper quartile (P = 0.032). An analysis of highly polymorphic SNPs indicated that this association is unlikely to be due to population substructure. We conclude that homozygosity for the C allele of FcepsilonRI alpha chain variant is associated with lower IgE levels.

Asthma↗
PubMed2002

Localization of thymidine phosphorylase expression in colorectal carcinoma tissues by in situ RT-PCR assay.

Abundant expression of thymidine phosphorylase (TP) mRNA and protein was observed in human carcinoma tissues relative to the corresponding normal tissues. However, the distribution of cells that express TP has not yet been determined. In this study, we examined enzyme activity of TP in 24 primary colorectal carcinoma tissues and the corresponding normal tissue. Using in situ reverse-transcription polymerase chain reaction (RT-PCR), a new methodology to determine the localization of genes of interest objectively and clearly, cell distribution was compared. The expression of TP mRNA is observed in normal infiltrating cells adjacent to colon carcinoma cells by in situ RT-PCR. Similar to the protein activity, the average TP mRNA expression (amplified products/x400 field) in cases of Dukes C and D is 76.8, while that of Dukes A and B is 42.5, being significantly different (p = 0.001). Moreover, the number of amplified products in colon carcinoma tissues showed a significant correlation with the enzyme activity evaluated by high-performance liquid chromatography (p < 0.01, R = 0.76). In conclusion, the expression of TP mRNA is localized in stromal tissues as determined by in situ RT-PCR and can reflect the level of TP enzyme activity.

Adenocarcinoma↗
PubMed2002

Relation between transverse conduction capability and the anatomy of the crista terminalis in patients with atrial flutter and atrial fibrillation: analysis by intracardiac echocardiography.

Although crista terminalis (CT) has been identified as the barrier to transverse conduction during typical atrial flutter (AFL), the relation between transverse conduction capabilities and anatomy of the CT remains unclear. The aim of the study was to evaluate that relation using intracardiac echocardiography (ICE). Ten patients with typical AFL (group AFL), 7 patients with paroxysmal atrial fibrillation (PAF) (group AF) and 8 patients without PAF or AFL (group N) underwent electrophysiologic testing. Using ICE images, the maximum diameter of the short axis of the CT (dCT) was measured and mapping and pacing catheters were positioned precisely. From extrastimulation delivered 1-2 cm anteriorly (free wall) or posteriorly (posterior wall) to the CT, the effective refractory period (CT-ERP) was determined as the longest coupling interval that resulted in split potentials at the mapping catheter positioned along the CT, a finding consistent with a transverse conduction block at the CT. The dCT was greater in group AFL than in groups AF and N (5.0+/-0.8 vs 4.3 +/-0.7, p<0.05 and 4.2+/-0.4 mm, p<0.01, respectively). The CT-ERP was longer during pacing from the posterior wall than from the free wall (307+/-68 vs 266+/-29 ms, p<0.05) as a whole group. The CT-ERP for the posterior wall pacing was longer in group AFL than in group N (339+/-80 vs 255+/-13, p<0.05). CT-ERP did not correlate with dCT; however, dCT was greater in patients with split potentials at the CT than in patients without them (4.9 +/-0.8 vs 4.1+/-0.5 mm, p<0.05). Therefore, the transverse conduction block of CT was more likely to occur in a thick CT. A limited transverse conduction capability of the CT is related to its thickness and might contribute to the development of typical AFL.

Adult↗
PubMed2002

Effects of a novel class III antiarrhythmic agent, NIP-142, on canine atrial fibrillation and flutter.

The effects of a new benzopyran derivative, NIP-142, on atrial fibrillation (AF) and flutter (AFL) and on electrophysiological variables were studied in the dog. NIP-142 (3mg/kg) was administered intravenously to pentobarbital-anesthetized beagles during vagally-induced AF and during AFL induced after placement of an intercaval crush. Isolated canine atrial tissues were studied using standard microelectrode technique. NIP-142 terminated AF in 5 of 6 dogs after an increase in fibrillation cycle length (CL) and prevented reinitiation of AF in all 6 dogs. NIP-142 terminated AFL in all 6 dogs without any appreciable change in flutter CL, and prevented reinitiation of AFL in all 6 dogs. NIP-142 prolonged atrial effective refractory periods (11+/-5%, 3+/-3%, 12+/-3%, and 10+/-5% from the baseline value at basic CLs of 150, 200, 300, and 350ms, respectively) without changes in intraatrial conduction time. The prolongation of the atrial effective refractory period was greater in the presence of vagal stimulation. NIP-142 decreased action potential phase-1 notch and increased phase-2 plateau height without making any changes in the action potential duration, although it did reverse carbachol-induced shortening of the action potential duration. In conclusion, NIP-142 is effective in treating AFL and vagally-induced AF by prolonging atrial refractoriness.

Animals↗
PubMed2002

Double-blind placebo-controlled trial of aprindine and digoxin for the prevention of symptomatic atrial fibrillation.

A multicenter, placebo-controlled, randomized, double-blind trial compared the preventive effect of aprindine and digoxin on the recurrence of atrial fibrillation (AF) with placebo, and also compare the effectiveness of these 2 drugs in the prevention of AF. Patients with symptomatic paroxysmal or persistent AF who had converted to sinus rhythm (SR) were randomly assigned aprindine (40 mg/day), digoxin (0.25 mg/day) or placebo and followed up on an outpatient basis every 2 weeks for 6 months. Of the 141 patients from 36 participating centers, 47 were given aprindine, 47 digoxin, and 47 were on placebo. After the 6-month follow-up, the Kaplan-Meier estimates of the percentage of patients remaining free of recurrent symptomatic AF on aprindine, digoxin and placebo were 33.3%, 29.2% and 21.5%, respectively. In patients remaining in SR for 15 days after from the start of follow-up, freedom from recurrence was significantly more prevalent in the aprindine group than in the placebo group (p=0.0414), but there was no significant difference between the digoxin and placebo groups. The rate of adverse events was similar in the 3 groups. In conclusion, neither aprindine nor digoxin had a significant effect on preventing relapse of symptomatic AF; however, recurrence of AF occurred later with aprindine than with placebo or digoxin.

Aged↗