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Hiroshi Inoue

Publications and source records attributed to Hiroshi Inoue.

At least 199 records · Page 11Linked to original sources

Transcriptional program of the endocrine pancreas in mice and humans.

The Endocrine Pancreas Consortium was formed in late 1999 to derive and sequence cDNA libraries enriched for rare transcripts expressed in the mammalian endocrine pancreas. Over the past 3 years, the Consortium has generated 20 cDNA libraries from mouse and human pancreatic tissues and deposited >150,000 sequences into the public expressed sequence tag databases. A special effort was made to enrich for cDNAs from the endocrine pancreas by constructing libraries from isolated islets. In addition, we constructed a library in which fetal pancreas from Neurogenin 3 null mice, which consists of only exocrine and duct cells, was subtracted from fetal wild-type pancreas to enrich for the transcripts from the endocrine compartment. Sequence analysis showed that these clones cluster into 9,464 assembly groups (approximating unique transcripts) for the mouse and 13,910 for the human sequences. Of these, >4,300 were unique to Consortium libraries. We have assembled a core clone set containing one cDNA for each assembly group for the mouse and have constructed the corresponding microarray, termed "PancChip 4.0," which contains >9,000 nonredundant elements. We show that this PancChip is highly enriched for genes expressed in the endocrine pancreas. The mouse and human clone sets and corresponding arrays will be important resources for diabetes research.

Animals↗

[Examination of measurement and its method of compensation of the sensitivity distribution using phased array coil for body scan].

The influence on the quality of images by measurement of a sensitivity distribution and the use of a sensitivity compensation filter was considered using an opposite-type phased array coil and volume-type phased array coil. With the opposite-type phased array coil, the relation between coil interval and filter was investigated for the IIC filter, SCIC filter (GE), and the Normalize filter (SIEMENS). The SCIC filter and Normalize filter showed distance dependability over the coil interval of SNR and uniformity was observed, and the existence of an optimal coil interval was suggested. Moreover, with the IIC filter, distance dependability over a coil interval was small, and the decrease in contrast with use was remarkable. On the other hand, with the volume-type phased array coil, the overlap of an array element was investigated to determine the influence it had on sensitivity distribution. Although the value stabilized in the arrangement center of an array element, and its near, was shown when volume-type phased array coil was used, SNR and uniformity decreased as it separated from the center. This showed that overlap of an array element does not have an adverse influence on image quality when the already optimized arrangement is used.

Image Enhancement↗

Clinical significance of the expression of activin A in esophageal carcinoma.

Activin A is a member of the transforming growth factor beta (TGF-beta) superfamily and is a strong differentiation factor of embryonic stem (ES) cells. It is unknown whether activin A has any correlation with carcinoma cell differentiation. We investigated the expression of activin-betaA (Act-betaA) which is a subunit of activin A, its receptor type I and IIb (ActRI, ActRIIb) and its inhibitor, inhibin-alpha (Inh-alpha), which is a subunit of inhibin A in esophageal carcinoma by reverse transcription polymerase chain reaction (RT-PCR) method. Act-betaA was overexpressed in carcinoma tissues significantly (p=0.030). On the other hand, Inh-alpha, ActRI and ActRIIb were neither overexpressed, nor suppressed. In immunohistochemistry and in situ hybridization analysis, Act-betaA expression was mainly derived from carcinoma cells. The mRNA expression of Act-betaA was not associated with carcinoma cell differentiation but lymph node metastasis (n0, 1, 2 vs. n3, 4; p=0.013) and clinical stage (I, II, III vs. IV; p=0.026). Moreover, patients with high mRNA expression of Act-betaA had a tendency to show poor prognosis compared to those with low mRNA expression (p=0.064). The finding indicated that activin A expression might not be associated with carcinoma cell differentiation but tumor aggressiveness such as lymph node metastasis in esophageal carcinoma.

Activin Receptors↗

Stimulation of CD40 inhibits Fas- or chemotherapy-mediated apoptosis and increases cell motility in human gastric carcinoma cells.

The widespread expression of CD40, a member of the tumor necrosis factor (TNF) receptor (TNFR) superfamily, is likely to account for the central role of CD40 in the regulation of humoral immunity and host defense. Interestingly, the expression of the CD40 in various types of carcinoma cells was often observed and conveys signals regulating diverse cellular responses, ranging from proliferation to growth suppression. Thus, the biologic role of the CD40-CD40L interaction in solid tumors is still controversial. In this study, we investigated the expression and function of the CD40 in gastric carcinoma cells. In 3-4,5 dimethylthiozol-2-yl-2,5-diphenyl tetrazolium bromide (MTT) assay and Annexin V/propidium iodide staining, CD40 stimulation using a soluble form of CD40 ligand did not affect cell viability, but significantly inhibited Fas-mediated or chemotherapy-mediated apoptosis in three CD40-positive gastric cancer cell lines. Moreover, in migration assay, CD40 stimulation induced an elevation of cell motility in CD40-positive gastric carcinoma cells. Our results show that the CD40 expression on gastric carcinoma makes cells less vulnerable to apoptosis induced by Fas or chemotherapy. These results suggest that the CD40 expression on gastric carcinoma may be associated with cell survival and elevation of cell motility.

Antineoplastic Agents, Phytogenic↗

[Susceptibilities of bacteria isolated from patients with lower respiratory infectious diseases to antibiotics (2001)].

From October 2001 to September 2002, we collected the specimen from 370 patients with lower respiratory tract infections in 16 institutions in Japan, and investigated the susceptibilities of the isolated bacteria to various antibacterial agents and antibiotics and patients' characteristics. Of 458 strains that were isolated from specimen (mainly from sputum) and assumed to be bacteria causing in inflammation, 456 strains were investigated. The breakdown of the isolated bacteria were: Staphylococcus aureus 69, Streptococcus pneumoniae 72, Haemophilus influenzae 85, Pseudomonas aeruginosa (non-mucoid) 44, P. aeruginosa (mucoid) 13, Klebsiella pneumoniae 32, Moraxella subgenus Branhamella catarrhalis 32, and others. Of 69 S. aureus strains, those with 4 micrograms/mL or more of MIC of oxacillin (methicillin-resistant S. aureus: MRSA) occupied 43.5%. Vancomycin and arbekacin showed the most potent activities against MRSA as observed in 2000. The frequency of S. pneumoniae exhibiting low sensitivity to penicillin (penicillin-intermediate S. pneumoniae: PISP + penicillin-resistant S. pneumoniae: PRSP) was 59.7% and both rates of PISP and PRSP were the highest after 1992. Carbapenems had strong activities against S. pneumoniae. Especially, panipenem and imipenem inhibited the growth of all 72 strains at 0.125 and 0.5 microgram/mL, respectively. Generally, all drugs had strong activities against H. influenzae with MIC90s of 16 micrograms/mL or less. The drug that had the strongest activity against H. influenzae was levofloxacin, which inhibited the growth of 80 of the 85 strains at 0.063 microgram/mL. Against P. aeruginosa mucoid strain, meropenem had a strong activity with MIC90 of 0.5 microgram/mL while, against non-mucoid strain, tobramycin had a strong activity with MIC90 of 2 micrograms/mL. K. pneumoniae showed good susceptibilities to all drugs except ampicillin and minocycline, and the MIC90s were 4 micrograms/mL or less. Particularly, cefmenoxime, cefpirome, and imipenem had the strongest activity (MIC90: 0.125 microgram/mL), and cefozopran had a strong activity, inhibiting the growth of all strains at 0.25 microgram/mL. Also, all drugs generally had strong activities against M. (B.) catarrhalis. MIC90s of all drugs were 4 micrograms/mL or less. The drug that had the strongest activity was minocycline and levofloxacin inhibiting all 32 strains at 0.063 microgram/mL. Most of the patients with respiratory infection were aged 70 years or older, accounting for approximately a half of the total (40.5%). As for the incidence by the diseases, bacterial pneumonia and chronic bronchitis were the highest, being noted in 39.2% and 37.3% of all the patients, respectively. The bacteria frequently isolated from the patients with bacterial pneumonia were S. aureus (19.3%) and S. pneumoniae (19.9%). In contrast, H. influenzae (22.0%) were frequently isolated from the patients with chronic bronchitis. Before the drug administration, the bacteria frequently isolated from the patients were S. pneumoniae (20.8%) and H. influenzae (21.5%). S. pneumoniae and H. influenzae decreased after the initiation of drug administration while S. aureus increased. The isolation frequency of P. aeruginosa was higher after than before the initiation of drug administration. The bacteria were frequently isolated from the patients who had already treated with cephems were S. aureus and P. aeruginosa. From the patients who had already treated with macrolides, S. pneumoniae was the most frequently isolated while S. aureus was the most frequently isolated from the patients pre-treated with quinolones.

Adult↗

Degradation profile of mRNA in a dead rat body: basic semi-quantification study.

To profile postmortem degradation of mRNA, total RNA was extracted, at given postmortem intervals, from the brain, lung, heart and liver of rats left at 20 degrees C. In electrophoretic analysis, total RNA was most stable in the brain, moderately stable in the lung and heart, and most unstable in the liver. Northern blot analysis of total RNA extracts from the brain and liver of dead rats with a cDNA probe for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) showed that GAPDH mRNA degraded in a similar fashion to total RNA. Analysis of the postmortem degradation profile of GAPDH mRNA with real-time reverse transcriptase-polymerase chain reaction (real-time RT-PCR) gave results consistent with those above, indicating that real-time RT-PCR is reliable for estimation of the mRNA level in specimens from dead bodies. Real-time RT-PCR analysis showed that degradation rates of three housekeeping genes, GAPDH, beta-actin and hypoxanthine guanine phosphoribosyltransferase, in the brains of dead rats were similar. The degradation rate of interleukin-1beta (IL-1beta) mRNA induced by intravenous injection of LPS to rats was higher than that of GAPDH mRNA in the lung. In real-time RT-PCR analysis using GAPDH mRNA as an internal standard, the detection level of IL-1beta mRNA decreased in the postmortem interval. However, enhanced expression of IL-1beta was detected for at least 3 days postmortem.

Actins↗

Oxygen affinity of hemoglobin regulates O2 consumption, metabolism, and physical activity.

The oxygen affinity of hemoglobin is critical for gas exchange in the lung and O(2) delivery in peripheral tissues. In the present study, we generated model mice that carry low affinity hemoglobin with the Titusville mutation in the alpha-globin gene or Presbyterian mutation in the beta-globin gene. The mutant mice showed increased O(2) consumption and CO(2) production in tissue metabolism, suggesting enhanced O(2) delivery by mutant Hbs. The histology of muscle showed a phenotypical conversion from a fast glycolytic to fast oxidative type. Surprisingly, mutant mice spontaneously ran twice as far as controls despite mild anemia. The oxygen affinity of hemoglobin may control the basal level of erythropoiesis, tissue O(2) consumption, physical activity, and behavior in mice.

Animals↗

Type III intestinal metaplasia and carcinoma express an identical carbohydrate-epitope revealed by a novel monoclonal antibody (2D11).

A monoclonal antibody (2D11, IgG2b) obtained by immunizing mice with a mucin fraction of the human gastric mucosa reacted specifically to intestinal metaplasia of human gastric mucosa and fetal intestinal mucosa but not to normal adult gastric, small intestinal or colonic mucosa in immunohistochemical staining. The results of Western blotting indicated that 2D11 recognized the high molecular weight glycoprotein(s) (mucin) of the stomach. Treatment of the antigens with sodium periodate abolished their reactivity to 2D11, and digestion of the antigens with beta-galactosidase reduced their reactivity to 2D11. Digestion of the antigens with pronase had no effect, however, suggesting that 2D11 recognizes the oligosugar moiety but not the peptide moiety of the antigens. Further immunohistochemical investigation showed that the reactivity of 2D11 was restricted to the Type IotaIotaIota intestinal metaplasia that is identified by a characteristic staining pattern with the high iron diamine-Alcian blue stain. 2D11 also reacted in high frequency to adenocarcinomas of the stomach (66.7%), pancreas (66.7%) and gallbladder (50.0%), but in low frequency to those in lung (8.3%) and colon (11.1%). It is of interest that 2D11 reacted to very restricted regions of the gastric adenocarcinomas. All monoclonal antibodies to mucin polypeptides (MUC1, 2, 3, 5AC and 6) examined stained intestinal metaplasia and carcinomas in a different pattern from 2D11 in immunohistochemistry. These facts indicate that Type IotaIotaIota intestinal metaplasia and carcinomas express carbohydrate chains identical to those expressed in the fetal intestinal mucosa, suggesting that both of them are closely related to fetal intestinal mucosa.

Animals↗

Neuropilin 1 and neuropilin 2 co-expression is significantly correlated with increased vascularity and poor prognosis in nonsmall cell lung carcinoma.

BACKGROUND: Cell-retained isoforms of vascular endothelial growth factor A (VEGF-A) have been reported to play an essential role in tumor progression through stromal neovascularization in malignant solid tumors. While more than 95% of nonsmall cell lung carcinoma (NSCLC) expresses cell-retained VEGF-A isoform, the clinicopathologic implications of neuropilin (NRP), considered the specific receptor for limited types of VEGF-A isoform, are not well understood. METHODS: The authors examined NRP1 and NRP2 mRNA expression in 68 NSCLCs and 15 extraneoplastic tissues by a densitometry-assisted, semi-quantitative reverse transcription-polymerase chain reaction. The authors determined the distinct expression of NRPs using the expression level of NRPs relative by optical density to beta2-microglobulin. The authors also investigated VEGF-A isoforms, their receptors, and the clinical implications. Vascularity of NSCLC was morphologically estimated on sections immunostained with anti-CD34 antibody. RESULTS: Eleven of 15 extraneoplastic specimens showed NRP1 expression (73.3%) and 8 showed NRP2 expression (53.3%). The expression level of NRP1 or NRP2 of neoplasmic tissue was higher than that of extraneoplastic tissues (P < 0.01, Mann-Whitney U test). Fifty-five and 44 NSCLCs expressed NRP1 and NRP2, respectively. Forty patients co-expressing NRP1 and NRP2 showed significantly poorer prognosis and increased vessel counts as compared to those 28 cases without co-expression (P < 0.05, log-rank test; P < 0.05, Mann-Whitney U test). CONCLUSIONS: The co-expression of NRP1 and NRP2 genes is significantly correlated with tumor progression through neovascularization in NSCLC. These results suggest that both NRP1 and NRP2 are key molecules for stromal vascularization by cell-retained VEGF in NSCLC.

Carcinoma, Non-Small-Cell Lung↗

Production and characterization of a monoclonal antibody for sweat-specific protein and its application for sweat identification.

Identification of body fluids is a common task in medico-legal practice, but specific markers for sweat have not been identified to date. To develop a method for identification of sweat, we identified a sweat-specific protein and produced monoclonal antibodies by immunizing mice with sweat proteins fractionated by anion-exchange chromatography. Among many sweat-reactive monoclonal antibodies obtained, one monoclonal antibody (G-81) was selected because of its unique specificity. G-81 reacted to sweat but not to other body fluids (e.g. serum, saliva, semen, milk, urine and tears) in ELISA. G-81 specifically stained the eccrine sweat gland and did not stain any other tissue including the apocrine sweat gland. In western blotting, G-81 reacted strongly to a 7 kDa band and faintly to 20, 27 and 33 kDa bands of sweat protein. The N-terminal amino acid sequence (18 amino acids) of G-81-reactive peptides was determined, and an identical sequence was found in an antibiotic peptide dermcidin (110 amino acids) reported recently, suggesting that G-81 recognized a fragment of dermcidin. The G-81-reactive peptide could be detected in 8,192-fold dilutions of sweat by ELISA and could be detected in 200-fold diluted sweat samples ( n=26) independent of the protein concentration. The G-81-reactive peptide was very stable and was able to detect sweat stains left for at least 11 weeks at room temperature without substantial loss of reactivity. These facts suggest that G-81 is a very useful tool for sweat identification in medico-legal practice.

Amino Acid Sequence↗

Intratumoral injection of dendritic cells after treatment of anticancer drugs induces tumor-specific antitumor effect in vivo.

We investigated the in vivo antitumor effects of intratumoral (i.t.) administration of dendritic cells (DC) after low-dose chemotherapy using cisplatin + 5-FU. Combination of i.t. injection of DC and systemic chemotherapy induced complete rejection of the treated tumor, MC38 murine adenocarcinoma. Furthermore, the antitumor effects were also observed on a distant tumor inoculated in the contralateral flank of the animal. When 10x the number of tumor cells were inoculated, the antitumor effect of the combination of DC after chemotherapy was also confirmed and in comparison to that of DC or chemotherapy alone, thereafter contributed to a greater prolongation of survival. To analyze the mechanisms of the systemic antitumor effect generated in this system, we assessed the cytolytic activity against inoculated tumors. The cytolytic activity of effector cells from treated animals was shown to be tumor-specific and was mainly CD8 and MHC Class-I (p < 0.01) restricted. CD4 and MHC Class-II treatment marginally inhibited the cytolytic activity but not significantly (p = 0.07, 0.08 respectively). The cytolysis of effector cells was enhanced more significantly by the treatment of both DC and chemotherapy, than that of either DC or chemotherapy alone. Our study suggests that the strategy of i.t. injection of DC after low-dose chemotherapy could be a powerful weapon to treat patients with cancer in the clinical settings.

Adenocarcinoma↗

Clinical and biological significance of S-phase kinase-associated protein 2 (Skp2) gene expression in gastric carcinoma: modulation of malignant phenotype by Skp2 overexpression, possibly via p27 proteolysis.

Reduced expression level of p27, a cyclin-dependent kinase inhibitor, is associated with high aggressiveness and poor prognosis of various malignant tumors, including gastric carcinoma. S-phase kinase-associated protein 2 (Skp2), a member of the F-box family of substrate-recognition subunits of Skp1-Cullin-F-box ubiquitin-protein ligase complexes, is necessary for p27 ubiquitination and degradation. In the present study, we examined the clinical and biological significance of Skp2 expression in human gastric carcinoma and the relationship between the expression of Skp2 and p27. Northern blot analysis showed that Skp2 mRNA was overexpressed in carcinoma tissues (P < 0.05), and the high Skp2 expression group showed significantly poorer prognosis in 98 patients with gastric carcinoma (P < 0.05). Immunohistochemical analysis showed that Skp2 protein was expressed predominantly in carcinoma cells. We also found an inverse correlation between the expression of Skp2 mRNA and p27 protein in vivo (P < 0.01). To analyze the biological behavior of Skp2, we established stably Skp2-transfected gastric carcinoma cell lines. Western blot analysis showed that Skp2-transfected cells expressed lower levels of p27 protein than the control cells. Skp2-transfected cells showed significantly higher levels of growth rate (P < 0.05), percentage of bromodeoxyuridine-positive cells after serum starvation (P < 0.01), resistance to apoptosis induction by actinomycin D treatment (P < 0.05), and invasion potential (P < 0.01) than the control cells. These findings indicate that Skp2 expression can modulate the malignant phenotype of gastric carcinoma, possibly via p27 proteolysis. Skp2 can play an important role in gastric carcinoma progression and would be a novel target for the treatment of gastric carcinoma as well as a strong prognostic marker.

Antibiotics, Antineoplastic↗

Expression of chemokine receptor CCR7 is associated with lymph node metastasis of gastric carcinoma.

The interactions of chemokine receptor CCR7 and its ligands are essential for migration of lymphocytes and dendritic cells to lymph nodes. In this study, we found that 4 of 6 (67%) gastric carcinoma cell lines tested expressed functional CCR7 for the chemokine CCL21/6Ckine, as demonstrated by calcium mobilization and actin polymerization assays. Moreover, we also showed that signaling through CCR7 induced chemotactic and invasive responses in CCR7-positive gastric carcinoma cells. In clinical samples, immunohistochemical assay showed that CCR7-positive carcinoma cells were detected in 42 of 64 (66%) cases and a significant difference in both lymph node metastasis (P < 0.001) and lymphatic invasion (P < 0.001) between CCR7-positive and -negative cases. Patients with CCR7-positive tumors had a significantly poorer prognosis than those with CCR7-negative tumors (P < 0.05). Stepwise regression analysis revealed that the most important factor related to lymph node metastasis was the expression of CCR7. These results indicated that CCR7 and its ligands interaction is associated with preferential lymph node metastasis of gastric carcinoma.

Actins↗

Rhodacyanine dyes as antimalarials. 1. Preliminary evaluation of their activity and toxicity.

The rhodacyanine dye MKT-077 (1), a potent antitumor agent, was found to possess strong in vitro activity against Plasmodium falciparum and a low cytotoxicity. Several new rhodacyanine dyes related to 1, containing a variety of linked heterocyclic moieties, were synthesized, and their antimalarial potencies were evaluated. The synthetic rhodacyanines were found to have EC(50) values against P. falciparum in vitro in the range of 4-300 nM. Several compounds in this series have remarkable selective toxicity profiles (>100).

Animals↗

Cardiac sympathetic denervation modulates the sympathoexcitatory response to acute myocardial ischemia.

OBJECTIVES: This study was designed to elucidate the influence of cardiac sympathetic denervation on the sympathoexcitatory response to acute myocardial ischemia during balloon coronary occlusion (BCO) in humans. BACKGROUND: Alterations of cardiac sympathetic nerve function could modulate sympathetic reflexes originating from the ischemic area. METHODS: In 23 patients with angina pectoris, we quantified the baseline cardiac sympathetic denervation of the ischemia-related area by iodine-123 metaiodobenzylguanidine ((123)I-MIBG), and transient changes in sympathetic activity during BCO by wavelet analysis of RR interval variability. RESULTS: Balloon coronary occlusion resulted in a transient augmentation of low-frequency (LF: 0.04 to 0.14 Hz) spectral components of RR interval variability in 4 of 12 patients with cardiac denervation and in 8 of 11 patients without denervation (p < 0.01 by the chi-square test). Consequently, the increase in LF components was significantly less during BCO in patients with cardiac denervation (34%) than in those without denervation (273%) (interaction: p < 0.05). In seven patients with severe ischemia provoked by a fall of > or = 10% in the left ventricular ejection fraction, LF components increased by 506% during BCO, regardless of the condition of cardiac denervation. In contrast, in patients with mild ischemia provoked by a fall of <10% in the ejection fraction, changes of LF components during BCO were significantly less in patients with denervation than in those without denervation (84 vs. 344%, p < 0.05). CONCLUSIONS: These findings suggest that if the provoked ischemia is not severe, cardiac sympathetic denervation could prevent ischemia-induced sympathoexcitation.

3-Iodobenzylguanidine↗

Modulation of the visual word retrieval system in writing: a functional MRI study on the Japanese orthographies.

We used functional magnetic resonance imaging (fMRI) to examine whether the act of writing involves different neuropsychological mechanisms between the two script systems of the Japanese language: kanji (ideogram) and kana (phonogram). The main experiments employed a 2 x 2 factorial design that comprised writing-to-dictation and visual mental recall for kanji and kana. For both scripts, the actual writing produced a widespread fronto-parietal activation in the left hemisphere. Especially, writing of kanji activated the left posteroinferior temporal cortex (lPITC), whereas that of kana also yielded a trend of activation in the same area. Mental recall for both scripts activated similarly the left parietotemporal regions including the lPITC. The writing versus mental recall comparison revealed greater activations in the left sensorimotor areas and right cerebellum. The kanji versus kana comparison showed increased responses in the left prefrontal and anterior cingulate areas. Especially, the lPITC showed a significant task-by-script interaction. Two additional control tasks, repetition (REP) and semantic judgment (SJ), activated the bilateral perisylvian areas, but enhanced the lPITC response only weakly. These results suggest that writing of the ideographic and phonographic scripts, although using the largely same cortical regions, each modulates the visual word-retrieval system according to their graphic features. Furthermore, comparisons with two additional tasks indicate that the activity of the lPITC increases especially in expressive language operations regardless of sensory modalities of the input stimulus.

Acoustic Stimulation↗