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Biomedical subjects

H Inoue

Publications and source records attributed to H Inoue.

At least 685 records · Page 38Linked to original sources
PubMed1997

5,6-Cis-penems: broad-spectrum anti-methicillin-resistant Staphylococcus aureus beta-lactam antibiotics.

5,6-cis-Penem derivatives have been synthesized and evaluated as anti-MRSA antibiotics. The cis-penems 5 and 6 showed potent activities against not only MRSA but also a wide variety of bacteria including beta-lactamase-producing microorganisms. These compounds were designed to have high affinity to the penicillin-binding protein 2a of MRSA and to form stable acyl intermediates with beta-lactamases by blocking the deacylating water molecule.

Anti-Bacterial Agents↗
PubMed1997

Suppressive effects of tranilast on the expression of inducible cyclooxygenase (COX2) in interleukin-1beta-stimulated fibroblasts.

We investigated the effects of tranilast on inducible cyclooxygenase (COX2)-mediated prostaglandin E2 (PGE2) production and enzyme induction in interleukin-lbeta (IL-1beta)-stimulated cultured dermal fibroblasts. IL-1beta enhanced PGE2 production in cultured fibroblasts. Tranilast did not affect constitutive cyclooxygenase (COX1) or COX2 activity in non-stimulated or IL-lbeta-stimulated fibroblasts. However, the COX2 expression induced by IL-1beta was inhibited by tranilast. This result, that IL-1beta-induced COX2 expression was suppressed by tranilast, was confirmed by immunohistochemical analysis. Thus, it is possible for tranilast to regulate PGE2 production by inhibiting COX2 induction.

Anti-Allergic Agents↗
PubMed1997

Purification of a regulatory subunit of type II cAMP-dependent protein kinase from Drosophila heads.

The cytosolic extract from Drosophila heads was separated using anion-exchange column chromatography. Two types of cAMP-dependent protein kinase (PKA), type I and type II, were detected, and type II PKA was found to be a major isozyme. The regulatory subunit of type II PKA (RII) was purified, and only one isoform was observed. The purified protein had an apparent molecular mass of 51 kDa on SDS gel electrophoresis. Partial amino acid sequences of the protein were almost identical with the RII alpha subunit of human. Since PKA has been implicated to be especially important for learning and memory in Drosophila, the RII subunit may play an essential role in the regulation of neuronal activity in the brain of Drosophila, and possibly in human.

Affinity Labels↗
PubMed1997

Inverse gene expression of prostacyclin and thromboxane synthases in resident and activated peritoneal macrophages.

Prostacyclin and thromboxane A2 produced from prostaglandin H2 are known to be important modulators with opposite biological activities. To examine possible roles of these prostanoids in immune responses, we have studied the gene expression of prostacyclin synthase (PGIS) and thromboxane synthase (TXS) in murine resident macrophages or in macrophages elicited with casein or bacillus Calmette-Guérin (BCG). Northern blot analyses showed that the PGIS mRNA was expressed in a decreasing order in the resident, and casein- and BCG-elicited macrophages. In contrast, the TXS mRNA was expressed in an increasing order in the resident, and casein- and BCG-elicited macrophages. On the other hand, the mRNA for cyclooxygenase-2, which produces PGH2 and participates in the production of prostanoids in inflammation, was expressed in both the resident and BCG-elicited macrophages but barely in the casein-elicited cells. In situ hybridization analysis showed that the expression of mRNAs for PGIS and TXS was ascribable not only to the alteration of the expression levels of both mRNAs in the each macrophage but also to the changes in subpopulations of the cells expressing these mRNAs. These observations suggested that the inverse gene expression of PGIS and TXS in macrophages contributes to immune responses by modulating the relative levels of prostacyclin and thromboxane A2.

Animals↗
PubMed1997

Human cholecystokinin type A receptor gene: cytogenetic localization, physical mapping, and identification of two missense variants in patients with obesity and non-insulin-dependent diabetes mellitus (NIDDM).

The human CCKAR gene was previously mapped to chromosome 4 using a panel of human/hamster somatic cell hybrids. We now report the cytogenetic and physical localization of the CCKAR gene. Using fluorescence in situ hybridization, we determined that CCKAR maps to 4p15.1-p15.2. On the physical map, CCKAR was adjacent to the marker AFMa283yh5, between AFMb355ya5 and WI-4086. A simple sequence repeat (D4S391) with high heterozygosity was found in the database, and CCKAR and this genetic marker were colocalized on two YACs (933D9 and 928A5). We also characterized the genomic structure and determined the exon-intron boundaries of the gene. This provided the opportunity to screen the gene in patients with non-insulin-dependent diabetes mellitus and/or obesity for single nucleotide changes using a single-strand conformational polymorphism strategy. Five sequence variants were identified in the coding sequence of the gene, including two missense variants (G21R and V365I). The results of these studies provide (1) precise genetic and physical mapping data, (2) exon-intron sequences for single nucleotide analysis, and (3) identification of two missense mutations in the CCKAR gene. The contribution of these CCKAR variants to normal physiology, to obesity, and to diabetes can now be evaluated.

Animals↗
PubMed1997

A mutant cell line resistant to Vibrio parahaemolyticus thermostable direct hemolysin (TDH): its potential in identification of putative receptor for TDH.

Thermostable direct hemolysin (TDH), a pore-forming toxin produced by Vibrio parahaemolyticus, is cytotoxic to Rat-1, a fibroblast cell line derived from rat embryo. Through mutagenesis of Rat-1 with nitrosoguanidine, we established a mutant cell line, MR-T1. MR-T1 was over 200 times more resistant to the cytotoxic activity of TDH than Rat-1. TDH increased membrane permeability of Rat-1 but not of MR-T1. Binding analysis showed that, while being able to bind to Rat-1. TDH failed to bind to MR-T1, indicating that MR-T1 is deficient in the putative receptor for TDH. Somatic hybrid cells between Rat-1 and MR-T1 were similarly sensitive to TDH as Rat-1. Moreover, TDH could bind to the hybrid cells as well as to Rat-1 cells. These results indicate that MR-T1 is promising for complementation cloning of a gene related to the putative receptor for TDH.

Animals↗
PubMed1997

alpha-2,3-Sialyltransferase type 3N and alpha-1,3-fucosyltransferase type VII are related to sialyl Lewis(x) synthesis and patient survival from lung carcinoma.

BACKGROUND: Biosynthesis of sialyl Lewis(x) (sLe(x)) requires a sialyltransferase for alpha-2,3-sialylation and a fucosyltransferase for alpha-1,3-fucosylation. To date, five human alpha-1,3-fucosyltransferase (Fuc-T) genes and five human alpha-2,3-sialyltransferase (ST) genes have been cloned. However, it is not known which enzyme is mainly responsible for sLe(x) synthesis. METHODS: Three hundred thirteen patients with nonsmall cell lung carcinoma who had a curative tumor resection were the subjects of this study. Using tumor tissues fixed in formaldehyde, amplification of genomic DNA of Fuc-T and ST was performed by PCR and correlated with sLe(x) staining and patient prognosis. RESULTS: The frequency of strong ST3N and Fuc-TVII amplification was significantly higher than that of STZ, ST4, Fuc-TIII, Fuc-TV, and Fuc-TVI amplification (P < 0.01). The frequency of sLe(x) staining was similar to ST3N and Fuc-TVII amplification. Survival of the patients whose tumors had strong amplification of both ST3N and Fuc-TVII was significantly shorter than that of patients whose tumors had no amplification of either gene (P < 0.01). In a multivariate analysis of survival, Fuc-TVII remained a statistically significant prognostic factor. CONCLUSIONS: In lung carcinoma, ST3N and Fuc-TVII may both be related to sLe(x) synthesis, and Fuc-TVII is a more important indicator of poor prognosis.

Adult↗
PubMed1997

Glucose-transporter-type-I-gene amplification correlates with sialyl-Lewis-X synthesis and proliferation in lung cancer.

Increased glucose transport is a common characteristic of most tumors. To examine the role of elevated glucose uptake in lung cancer, we performed PCR amplification of 2 facilitative glucose transporter genes (GLUT1 and GLUT3) and immunohistochemical staining for GLUT1, proliferating cell nuclear antigen (PCNA), and sialyl Lewis x (sLe(x)) on tumor specimens from 327 patients with lung cancer who underwent surgical resection from 1980 to 1993. To evaluate the relationship between GLUT, alpha-2,3-sialyltransferase (ST), and alpha-1,3-fucosyltransferase (Fuc-T) genes, PCR amplification of the ST3N and Fuc-TVII also was performed. Amplification of GLUT1 was significantly greater than that of GLUT3. GLUT1 and GLUT3 amplification correlated with PCNA staining (p < 0.01). In addition, GLUT1 amplification correlated with the grading of sLe(x) staining as well as with the grading of GLUT1 staining (p < .03, p < 0.01). GLUT1 was co-amplified with ST3N and Fuc-TVII genes, which are involved in the synthesis of sLe(x) (p < 0.01). The survival of patients whose tumors showed GLUT1 amplification was significantly shorter than that of patients whose tumors did not (p < 0.01). In a multivariate analysis of survival, GLUT1 remained a statistically significant prognostic factor. Our results suggest that GLUT1 amplification may participate in sLe(x) synthesis as well as in proliferation, and may be of prognostic value in lung cancer.

Adult↗
PubMed1997

Retinoids suppress phorbol ester-mediated induction of cyclooxygenase-2.

Cyclooxygenase-2 expression is up-regulated in transformed cells and tumors. Because this enzyme catalyzes the synthesis of prostaglandins, strategies aimed at suppressing its expression may prove useful in preventing or treating cancer. We investigated the ability of retinoids to suppress phorbol ester-mediated induction of cyclooxygenase-2 in human oral epithelial cells. Treatment with phorbol myristate acetate (PMA) resulted in approximately a 3-fold increase in the production of prostaglandin E2 (PGE2). Retinoids [all-trans-retinoic acid (RA), 13-cis-RA, and retinyl acetate] markedly suppressed PMA-mediated increases in amounts of cyclooxygenase-2 (Cox-2) and the production of PGE2. Retinoids also suppressed the induction of Cox-2 mRNA by PMA. Nuclear run-offs revealed increased rates of Cox-2 transcription after treatment with PMA; this effect was inhibited by all-trans-RA. Transient transfection experiments showed that PMA caused about a 2-fold increase in Cox-2 promoter activity, an effect that was suppressed by all-trans-RA. Our data indicate that treatment of oral epithelial cells with PMA is associated with enhanced transcription of Cox-2 and increased production of PGE2. These effects of PMA were inhibited by retinoids.

Anticarcinogenic Agents↗
PubMed1997

Isolation, characterization, and chromosomal mapping of the human Nkx6.1 gene (NKX6A), a new pancreatic islet homeobox gene.

Nkx6.1 (gene symbol NKX6A), a new member of the NK homeobox gene family, was recently identified in rodent pancreatic islet beta-cell lines. The pattern of expression suggested that this gene product might be important for control of islet development and/or regulation of insulin biosynthesis. We now report cloning of human NKX6A, characterization of its genomic structure, and its chromosomal localization. The predicted protein of human NKX6A contained 367 amino acids and had 97% identity to the hamster protein. The highly conserved NK decapeptide and homeodomain regions were identical between human and hamster, suggesting functional importance of these domains. The coding region spanned approximately 4.8 kb and was composed of three exons. The gene was localized to four CEPH "B" yeast artificial chromosome clones (914B4, 951G9, 981D6, and 847B3), and a nearby polymorphic marker (D4S1538) on chromosome 4 was identified < 1270 kb from the gene. Using fluorescence in situ hybridization, we also determined that NKX6A maps to 4q21.2-q22.

Amino Acid Sequence↗
PubMed1997

Structure and expression of the human FHIT gene in normal and tumor cells.

The FHIT gene, encoded by 10 exons in a 1.1-kb transcript, encompasses approximately 1 Mb of genomic DNA, which includes the hereditary RCC t(3;8) translocation break at 3p14.2, the FRA3B common fragile region, and homozygous deletions in various cancer-derived cell lines. Because some of these genetic landmarks (e.g., the t(3;8) break between untranslated FHIT exons 3 and 4, a major fragile region that includes a viral integration site between exons 4 and 5, and cancer cell homozygous deletions in intron 5) do not necessarily affect coding exons and yet apparently affect expression of the gene product, we examined the FHIT locus and its expression in detail in more than 10 tumor-derived cell lines to clarify mechanisms underlying aberrant expression. We observed some cell lines with apparently continuous large homozygous deletions, which included one or more coding exons; cell lines with discontinuous deletions, some of which included or excluded coding exons; and cell lines that exhibited heterozygous and/or homozygous deletions, by Southern blot analysis for the presence of specific exons. Most of the cell lines that exhibited genomic alterations showed alteration of FHIT transcripts and absence or diminution of Fhit protein.

Acid Anhydride Hydrolases↗
PubMed1997

Sensitive detection of human globin chains by microbore high-performance liquid chromatography and its forensic application.

This paper describes a simple and sensitive high-performance liquid chromatographic (HPLC) method for the detection of human globin chains in blood and bloodstains. The method involves direct injection of the filtered samples of dilute hemolysates or bloodstain extracts onto a microbore C4 reversed-phase column (2.1 mm I.D.) with UV detection at 220 nm. Microbore HPLC offers a significant improvement in sensitivity with little loss of the resolution of globin chains and only small variations in the determination of gamma chain composition. The detection limit of hemoglobin (Hb) was 0.1 microgram, which is equivalent to about 1 nl of fresh whole blood. Umbilical cord blood could be differentiated from adult blood in stains that were up to twenty weeks old, by the presence of gamma globin chains. The present method will be useful for detection of abnormal Hbs and for the determination of gamma chain composition in clinical laboratories, as well as in the practice of forensic science for the analysis of minute amounts of blood and bloodstains.

Blood Stains↗
PubMed1997

Ex vivo and in vivo evaluation of the blood compatibility of surface-modified polyurethane catheters.

Catheter model tubes were prepared from a medical-grade polyetherurethane and their outer surfaces modified by surface-graft polymerization of acrylamide and dimethyl acrylamide (DMAA). The surface-graft layer was characterized by means of dry staining, scanning electron microscopy (SEM), X-ray photoelectron spectroscopy, and protein adsorption. Ex vivo evaluation for the blood compatibility of the surface-modified polyurethane was carried out using the polyurethane tube as an arterio-venous shunt between the carotid artery and the jugular vein of rabbits. When the surface density of grafted polymer was in the range of 10-30 microg/cm2, the in vitro adsorption of IgG exhibited a minimum value and platelet adhesion to the grafted polyurethane surface was insignificant, in marked contrast with that to the virgin (nonmodified) surface. The in vivo blood compatibility of polyurethane was evaluated by implanting the catheter tube in the inferior vena cava of rabbits from the femoral vein after ligation of a distal site of the exposed femoral vein. After remaining there for predetermined periods of time, the implanted catheters were taken out together with the veins of the rabbits that had been heparinized and sacrificed just prior to excision of the veins. After exchange of the blood in the veins for saline, the excised veins were opened by cutting longitudinally to inspect for clot formations on the surfaces of the implanted catheters. Occlusion of the inferior vena cava was not observed for any of the catheters, nor was there any apparent damage or microembolizations in the lungs and kidneys. Many small-sized clots were observed on the surfaces of the nonmodified polyurethane tubes after a 2-week implantation whereas the catheter surfaces grafted with DMAA polymer chains had a much smaller number of clots. When the blood compatibility of polyurethane surfaces was graded for relative evaluation from one (marked clotting) to five (no clotting) based on the size and number of the clots, the evaluation results were as follows: 3.1 (virgin, 2 weeks), 4.0 (grafted, 1 week), 4.1 (grafted, 2 weeks), and 3.5 (grafted, 1 month).

Acrylamides↗
PubMed1997

Direct evidence of high DNA binding activity of transcription factor AP-1 in rheumatoid arthritis synovium.

OBJECTIVE: To investigate the possible activation of transcription factor AP-1 in rheumatoid arthritis (RA) and its involvement in the pathogenesis of RA. METHODS: Synovial tissues and peripheral blood samples were obtained from 25 patients with RA and 5 patients with osteoarthritis (OA) during arthroplasty and synovectomy. The synovial tissue was digested with collagenase and separated into adherent and nonadherent cells by plastic-adhesion methods. Nuclear extracts obtained from each sample were examined by electrophoretic mobility shift assay to determine the DNA binding activity of AP-1. The expression of c-fos and c-jun messenger RNA (mRNA) was examined by in situ reverse transcription assay. RESULTS: A markedly high DNA binding activity of AP-1 was detected in the synovial tissues of RA patients, while virtually no activity or only a little activity was observed in OA patients. Following separation of adherent and nonadherent cells, the AP-1 activity was mainly detected in adherent cells, which consisted of synovial cells and macrophages. However, the activity was significantly higher in the mononuclear cells infiltrating into RA synovium than in RA peripheral blood mononuclear cells. The high DNA binding activity of AP-1 in RA correlated with the expression of c-fos and c-jun mRNA in situ. Furthermore, AP-1 binding activity also correlated with disease activity. CONCLUSION: In RA synovium, AP-1 DNA binding activity was constitutively up-regulated. These findings suggest that AP-1 may play an important role in the pathogenesis of RA, including synovial hyperplasia and abnormal immune responses.

Aged↗
PubMed1997

Quantitative evaluation of blood flow distribution to exercising and resting skeletal muscles in patients with cardiac dysfunction using whole-body thallium-201 scintigraphy.

BACKGROUND AND HYPOTHESIS: Decreased blood flow to working muscles makes an important contribution to exercise intolerance in patients with chronic heart failure. This study was undertaken to examine whether maldistribution of skeletal muscle blood flow is closely related to exercise intolerance in patients with cardiac dysfunction. METHODS: Whole-body thallium scintigraphy was performed during one-leg exercise in 11 patients with left ventricular (LV) dysfunction (LV ejection fraction < 45%). Blood flow distribution to the exercising and resting legs was quantified by expressing regional thallium counts as a percentage of the whole-body counts at rest, at the level of anaerobic threshold, and at peak exercise. RESULTS: At anaerobic threshold, the thallium activity of exercising muscle increased from 4.2 +/- 0.7 to 14.0 +/- 2.5% (p < 0.05) in the thigh and from 1.7 +/- 0.3 to 4.1 +/- 0.9% (p < 0.05) in the calf, compared with the resting value. Consequently, the ratio of thallium activity between exercising and resting legs increased to 2.7 +/- 0.7 (p < 0.05) in the thigh and to 2.3 +/- 0.7 (p < 0.05) in the calf. When plotted as a function of anaerobic threshold, thallium activity of the exercising thigh (r = 0.78, p < 0.05) and the thallium ratio between exercising and resting thigh (r = 0.69, p < 0.05) declined with the reduction of exercise tolerance. These correlations were not observed in calves. CONCLUSION: Whole-body thallium scintigraphy demonstrated a maldistribution of leg blood flow in patients with reduced aerobic exercise capacity, suggesting that this abnormality could play an important role in exercising intolerance in these patients.

Adult↗
PubMed1997

Feasibility of the transparent cap-fitted colonoscope for screening and mucosal resection.

PURPOSE: A transparent plastic cap of 17 mm in outer diameter, 15 mm in inner diameter, and 10 mm in length can be easily attached to the tip of a colonoscope. By using the cap, a better view of the lesions hiding at the opposite side of the fold can be obtained. When a flat colonic lesion is found, a submucosal injection of saline solution is performed, the target mucosa is sucked inside the cap, snared under a full endoscopic suction, and resected by an electrical current. This procedure is called endoscopic mucosal resection using transparent cap-fitted endoscope (EMRC). Feasibility of the cap-fitted colonoscope for screening colonoscopic examination and mucosal resection was evaluated. MATERIALS AND METHODS: One hundred forty patients were randomly allocated for screening with a normal colonoscope (NCF) or that with the cap-fitted colonoscope (CCF). Average time for insertion up to the cecum, patients' discomfort during insertion expressed in 4 degrees, and average number of lesions found in one patient were compared. Thirty lesions randomly allocated for mucosal resection with conventional strip biopsy or EMRC were also evaluated. RESULTS: Time consumed for insertion up to the cecum with the CCF (12.4 +/- 6.6 minutes) was the same as that with the NCF (12.3 +/- 5.2 minutes), and there was no significant difference in patients' discomfort; however, the average number of lesions found in one patient was larger when using the CCF (0.86 +/- 0.96) than when using the NCF (0.58 +/- 0.81). For mucosal resection, 40 flat or wide-based lesions including 6 mucosal carcinomas were resected with EMRC. We experienced only one pinhole perforation of the ascending colon by heat damage, which was treated successfully by surgery. There was no other major complication or recurrence. CONCLUSION: The cap-fitted endoscope was equal in maneuverability, was excellent in sensitivity in comparison with the regular colonoscope, and was thought to be feasible both in screening and mucosal resection.

Aged↗