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Biomedical subjects

H Inoue

Publications and source records attributed to H Inoue.

At least 91 records · Page 5Linked to original sources
PubMed1997

Genetic and molecular characterization of Neurospora crassa mus-23: a gene involved in recombinational repair.

A newly isolated mutant, mus-23, of Neurospora crassa was found to be highly sensitive to a wide variety of mutagens, including UV light, methyl methanesulfonate, 4-nitroquinoline 1-oxide, N-methyl-N'-nitro-N-nitrosoguanidine and tert-butyl hydroperoxide. This mutant was originally isolated as a mutant that could not grow on medium containing histidine. Meiosis and sporulation were defective in homozygous crosses between mus-23 haploids. The mus-23 gene is located on the right arm of LGII, between fl and trp-3. Analyses of epistasis between mus-23 and other mutations that cause defects in DNA repair indicated that the mus-23 gene belongs to the same DNA repair group as mei-3, which is the Neurospora homolog of the Saccharomyces cerevisiae gene RAD51. The double mutant carrying mus-23 and uvs-3 mutations was lethal. The mus-23 gene was cloned by complementation of the MMS-sensitive phenotype of the mus-23 mutant. The gene contained an open reading frame of 1578 bp and did not contain any introns. The molecular weight of the predicted mus-23 gene product was 60.4 kDa. Computer analyses revealed that the MUS23 protein has significant homology to Mre11p, which is known to be involved in recombinational repair in S. cerevisiae. The level of mus-23 transcripts increased significantly within 60 min of treatment with UV or MMS and then gradually decreased. The role of MUS23 protein in recombinational repair is discussed.

Amino Acid Sequence↗
PubMed1997

Genetic studies of the leptin receptor gene in morbidly obese French Caucasian families.

Family studies have shown that in some populations up to 75% of the variation of body mass index can be explained by genetic factors. However, in humans, no major obesity gene has been identified to date. In contrast, there are a number of genetically well defined animal models for obesity. In two of those models (ob/ob and db/db), defects in the same pathway are responsible for obesity. Recently, some evidence has been found for the OB gene also being involved in human obesity. In this study we investigated the potential role of the OB receptor (OBR) in the etiology of massive obesity in humans using familial linkage analyses and case-control association studies. The typing of two microsatellite markers (D1S198 and D1S209), flanking the OBR gene, in 256 sib pairs showed no evidence for linkage with obesity. In order to be able to detect small gene effects, association studies with a 3'-UTR insertion/deletion polymorphism were carried out. The results of these analyses remained non-significant (chi 2 = 3.442, P = 0.18). However, subjects heterozygous for the insertion/deletion polymorphism showed a slight trend towards lower insulin values 30 min after an oral glucose load compared to homozygous individuals (P = 0.02). In summary, our results do not support a major role of the human OBR gene in the development of morbid obesity in our population.

Adult↗
PubMed1997

Soluble E-selectin, ICAM-1 and VCAM-1 levels in systemic and coronary circulation in patients with variant angina.

OBJECTIVE: The purpose of this study was to assess whether the plasma levels of soluble adhesion molecules including E-selectin, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) are elevated in patients with variant angina and whether they are released in the coronary circulation. METHODS: Antecubital venous plasma samples were collected from 33 patients with variant angina, 22 patients with stable effort angina and 20 control subjects. Samples were also collected from the aortic root (AO) and the coronary sinus (CS) in 18 patients with variant angina before and after left coronary spasm induced by intracoronary injection of acetylcholine. The soluble adhesion molecules were assayed by enzyme immunoassay. RESULTS: Antecubital venous plasma soluble E-selectin (P < .05), ICAM-1 (P < .01) and VCAM-1 (NS) levels were higher in the variant angina group than in the control group, respectively. The plasma soluble ICAM-1 level was also higher (P < .01) in the variant angina group than in the stable effort angina group. In the variant angina group, both soluble ICAM-1 (P < .05) and VCAM-1 (P < .01) levels were significantly lower in CS than AO at baseline. In contrast, after the spasm the plasma soluble ICAM-1 level was (P < .05) higher in CS than AO and the CS-AO differences of soluble ICAM-1 (P < .05) and VCAM-1 (P < .05) increased as compared with the baseline, respectively. These values were remained unchanged in the stable effort angina group after rapid atrial pacing and in the control group after administration of acetylcholine. CONCLUSIONS: Circulating plasma levels of both soluble E-selectin and ICAM-1 were elevated in patients with variant angina, indicating an association of an inflammatory reaction with coronary spasm. Both soluble ICAM-1 and VCAM-1 appeared to be trapped in the coronary circulation at baseline and released into the coronary circulation following coronary spasm and reperfusion in the patients.

Acetylcholine↗
PubMed1997

Effect of azelastine on PGE2 production in fibroblasts in normal skin. The possibility of inhibition of inducible cyclooxygenase by azelastine.

The effects of azelastine on prostaglandin E2 (PGE2) production were investigated by using cultured normal dermal fibroblasts obtained from the same traumatic region of 3 patients and the CRL-1475 cell line. Interleukin-1beta (IL-1) enhanced PGE2 production in cultured normal fibroblasts and CRL-1475 cells. 10(-6) M azelastine inhibited PGE2 production in these IL-1-stimulated fibroblasts. However, the drug did not influence spontaneous PGE2 production in cultured CRL-1475 fibroblasts not stimulated with IL-1 but slightly it increased in cultured normal dermal fibroblasts under the same conditions. These results suggest that azelastine either regulates synthesis of an inducible cyclooxygenose protein or inhibits PGE2 production as an inducible cyclooxygenase inhibitor.

Accidents↗
PubMed1997

Prevalence of the Trp64Arg missense mutation of the beta3-adrenergic receptor gene in Japanese subjects.

Prompted by the recent findings that a tryptophan to arginine (Trp64Arg) mutation in the beta3-adrenergic receptor gene was associated with an earlier onset of non-insulin-dependent diabetes mellitus (NIDDM) in Pima Indians, with abdominal obesity and insulin resistance in Finns, and with an increased capacity to gain weight in French whites, we studied the prevalence of this mutation in 231 diabetic and 95 nondiabetic Japanese subjects and assessed its contribution to the development of obesity and NIDDM. The allelic frequencies of the mutation were 0.18 in diabetic and 0.23 in nondiabetic subjects, showing no significant difference between the two groups (P = .067). In nondiabetic subjects, body mass index (BMI) did not differ between those with and without the mutation (22.2 +/- 3.5 v 21.4 +/- 3.2 kg/m2, P = .252). In NIDDM subjects, BMI at the time of study and maximal BMI before the start of treatment did not differ between those with and without the mutation (22.8 +/- 2.6 v 23.2 +/- 3.7 kg/m2, P = .678, and 24.7 +/- 2.6 v 24.9 +/- 3.1 kg/m2, P = .277). Homozygotes for the mutation did not have trends to have increased BMI in either diabetic or nondiabetic subjects. The age at diagnosis of NIDDM also did not differ between the two groups (48.8 +/- 9.9 v 47.8 +/- 12.5 years, P = .796). Fasting serum cholesterol and triglyceride levels and systolic and diastolic blood pressure before the start of treatment did not differ between NIDDM subjects with and without the mutation. In conclusion, although the Trp64Arg mutation is not uncommon in Japanese, it does not appear to be associated with obesity, NIDDM, age at diagnosis of NIDDM, or dyslipidemia. Our results suggest that the mutation has minor effects, if any, on the development of obesity and NIDDM in Japanese.

Adult↗
PubMed1997

A monoclonal antibody that recognizes ganglioside GD1b in the rat central nervous system.

A monoclonal antibody (MAb), generated by immunizing BALB/c mice with homogenized bovine retinal tissue, was specific to ganglioside GD1b incorporated into liposome membranes. The antibody (MAb-5G6), classified as IgM, immunostained intensely the perikaryon and processes of motoneurons in the cranial motor nuclei and spinal cord. Spinal and trigeminal ganglion cells were also immunopositive to the MAb. Some fiber tract systems, such as the spinal and mesencephalic trigeminal tracts, the solitary tract and the posterior funiculus, were also immunoreactive to the MAb. These findings suggest that MAb-5G6 labeled specifically neurons with axons extending outside of the central nervous system as a peripheral nerve. The immunoreactive substances were visualized under electron microscopy just beneath the postsynaptic membrane and just inside the plasmalemma of the thick dendrites. No axon terminal was immunolabeled by the MAb. In the rat embryos, immunoreactivity to MAb-5G6 was found in the dorsal and ventral root fibers on the 15th embryonic day (E15). However, cell bodies of the spinal ganglion cells and motoneurons were immunostained by MAb-5G6 at a later stage (E20). The ventral commissure fibers in the floor plate of the spinal cord were transiently immunolabeled during E13-15.

Animals↗
PubMed1997

Guyon's canal syndrome. A different clinical presentation caused by an atypical fibrous band.

An atypical case of Guyon's canal syndrome is reported. A 40-year-old woman with hypoaesthesia and claw-finger deformity of the little finger, hypothenar muscle atrophy but no first dorsal interosseous muscle atrophy, underwent release of a fibrous band to decompress the ulnar nerve at the deep branch just proximal to the branch to the abductor digiti minimi muscle. The claw-finger deformity recovered 1 week after surgery.

Adult↗
PubMed1997

The central slip attachment fracture.

Eight displaced central slip attachment fractures were treated by open reduction and internal fixation to avoid boutonniere deformity, to reduce the fracture anatomically and to allow early mobilization of the joint. This injury should be recognized as a disruption of the dynamic extensor mechanism associated with an intraarticular fracture, fracture-dislocation or soft tissue injury of the PIP joint. We have grouped central slip attachment fractures into three types according to the mechanism of injury, with suggested methods of treatment.

Adolescent↗
PubMed1997

Relationship between postoperative intraocular pressure elevation and residual sodium hyaluronate following phacoemulsification and aspiration.

PURPOSE: To investigate the relationship between postoperative intraocular pressure (IOP) elevation and the amount and viscosity of sodium hyaluronate remaining in the anterior chamber after phacoemulsification and aspiration (PEA). SETTING: Hachioji Medical Center and Tokyo Medical College Hospital, Tokyo, Japan. METHODS: In 107 eyes, washout (irrigation and aspiration [I/A]) of sodium hyaluronate was performed for randomly assigned durations of 5, 10, or 20 seconds following PEA and intraocular lens (IOL) implantation. At the conclusion of washout, the flow from the I/A tip was reversed and a sample of the irrigating solution obtained from the anterior chamber. The residual sodium hyaluronate concentration was measured by sandwich assay. The kinematic viscosity of the sample was calculated from a regression equation derived from measured viscosities of standard hyaluronate solutions of various concentrations. RESULTS: The preoperative mean IOP in the groups that received 5-, 10-, and 20-second washout was 11.9, 10.9, and 11.6 mm Hg, respectively. The mean IOP on the first postoperative day in the 5-second group was significantly higher (P = .001, Kruskal-Wallis test) than those in the 10- and 20-second groups (10.8 and 11.6 mm Hg, respectively). A significant correlation was observed between residual hyaluronate concentration and IOP on the first postoperative day (Spearman's rank correlation coefficient 0.319, n = 80, P = .045) and also between kinematic viscosity and IOP on the first postoperative day (one-factor analysis of variance, P < .001). CONCLUSION: The sodium-hyaluronate-induced IOP elevation is related to its viscosity as well as to its high molecular weight. Washout times of at least 10 seconds are desirable to prevent IOP elevation.

Aged↗
PubMed1997

Structural analysis of A-protein of cucumber green mottle mosaic virus and tobacco mosaic virus by synchrotron small-angle X-ray scattering.

The size and shape of A-protein of tobacco mosaic virus coat protein (TMVP) and cucumber green mottle mosaic virus coat protein (CGMMVP) were evaluated by means of small-angle X-ray scattering (SAXS) using a synchrotron radiation source, complemented by electron microscopic observations. The results imply that TMV and CGMMV A-proteins are composed of three and two subunits, respectively, stacked in the shape of an isosceles triangular prism at lower ionic strength. Considering the difference of the A-protein structure at higher and lower ionic strength, the globular core structure was proposed as a subunit which might be modeled as a thin isosceles triangular prism composed of four globular cores joined by rather flexible segments. These cores correspond probably to four helical regions in a subunit, and rearrange their relative positions according to the external conditions. A slight rearrangement of core positions in a subunit may result in the formation of A-proteins of various shapes.

Capsid↗
PubMed1997

Relationship between prognostic score and thyrotropin receptor (TSH-R) in papillary thyroid carcinoma: immunohistochemical detection of TSH-R.

We have demonstrated the expression of thyrotropin receptor (TSH-R) in thyroid neoplasms (13 adenomas, 21 papillary carcinomas, two follicular carcinomas) and adjacent normal thyroid using the monoclonal antibody against human TSH-R and have also demonstrated a relationship between prognostic scores and the expression of TSH-R. Among the adenomas, eight showed an intensity similar to that of normal thyroid and five showed a higher intensity than normal. Two tumours exhibited heterogeneous distribution of TSH-R. Among the papillary carcinomas, seven showed similar intensity to normal tissue and four showed higher intensity and ten showed weaker intensity. Eight tumours showed heterogeneous distribution of the stain. Among the follicular carcinomas, one showed similar intensity to normal tissue and the other exhibited weaker intensity. Both cases showed homogeneous distribution of TSH-R. The adenomas never showed a weaker intensity than normal thyroid, but various intensities of TSH-R occurred in differentiated carcinomas. There was no significant relationship between the clinical data and the signal intensity in the adenomas. Among the papillary carcinomas, however, the group with weaker intensity had significantly poorer prognostic scores than the other two groups. Thus, we assume that low TSH-R may be expressed by the clinically high-risk group of patients with papillary thyroid carcinoma.

Adenocarcinoma, Follicular↗
PubMed1997

Functional analysis of the evolutionary conserved arginine 182 residue in human glutathione S-transferase P1-1.

The mutational replacement of Arg182 with threonine markedly decreased the specific activities for GSH-conjugation reaction. The Kcat of R182T for GSH-[1-chloro-2,4-dinitrobenzene] conjugation reaction was about 100-fold smaller than that of the wild type. On the other hand, the affinity for GSH of R182T was not significantly affected. The pKa of the thiol group of GSH bound in R182T was approximately 0.9 pK units higher than those in the wild type, but the Kcat/Km1-chloro-2,4-dinitrobenzene values at high pH were not so much lower than those of the wild type. The thermostability of R182T was significantly lower than that of the wild type. Therefore, Arg182 seems to be important for the construction of the active enzyme structure.

Amino Acid Sequence↗
PubMed1997

Benzo[a]pyrene up-regulates cyclooxygenase-2 gene expression in oral epithelial cells.

Cyclooxygenase may be important in the pathogenesis of smoking-related cancer because it activates carcinogens and catalyzes prostaglandin biosynthesis. We determined the effects of benzo[a]pyrene (B[a]P), a polycyclic aromatic hydrocarbon in tobacco smoke, on cyclooxygenase-2 (Cox-2) mRNA, protein and synthesis of prostaglandin E2 (PGE2) in normal and transformed oral epithelial cells. Treatment with B[a]P caused a dose-dependent increase in production of PGE2, with a maximal increase of approximately 100%. Enhanced synthesis of PGE2 was associated with increased amounts of Cox-2 protein. B[a]P also caused a two-fold increase in Cox-2 mRNA in both normal and transformed cells. Transient transfections with a Cox-2 promoter construct showed that B[a]P-mediated induction of Cox-2 mRNA reflected increased transcription. Levels of Cox-1 were unaffected by B[a]P. B[e]P did not affect the synthesis of PGE2 or amounts of Cox-2. These data are important because B[a]P-mediated induction of Cox-2 may predispose to carcinogenesis by enhancing the production of mutagens and the synthesis of prostaglandins.

Benzo(a)pyrene↗
PubMed1997

Diversity of epitopes recognized by cytotoxic T lymphocytes that are specific for rejection antigen peptide pRL1a presented on BALB/c leukemia RL Male 1.

Cytotoxic T lymphocytes (CTL) generated in (BALB/c x C57BL/6)F1 (CB6F1) and BALB/c spleen cells stimulated with BALB/c radiation-leukemia RL Male 1 cells or pRL1a (IPGLPLSL) peptide itself recognized pRL1a on RL Male 1 in association with Ld. We first studied pRL1a peptide residues used for binding to the Ld molecule by examining the inhibition by variant peptides with single Ala substitutions at each position (P) of recognition of P815 target cells sensitized with Ld-binding p2Ca (LSPFPFDL) peptide for BALB/c anti-p2Ca CTL. The results showed that Leu at P8 is predominantly involved in the binding and Pro at P2 is partially involved. Substitution of Gly to Ala at P3 increased binding. We then investigated the epitope residues recognized by four pRL1a-specific CTL clones by examining their cytotoxicity against the P815 target sensitized with variant pRL1a peptides. Recognition by clone Y-16 involved predominantly Leu at P4 and P6, and also Pro at P5 and Ser at P7, and partially Ile at P1. Recognition by clone U-41 involved predominantly Ile at P1 and Leu at P6, and partially Gly at P3, Leu at P4, Pro at P5 and Ser at P7. Recognition by clone P-2 involved predominantly Leu at P4 and P6, and Ser at P7, with no partial involvement of other substitutions being observed. Finally, recognition by clone B-24 predominantly involved all residues, except Gly at P3, which was partially involved. TCR V beta genes utilized by those CTL clones were different. The findings show that tumor antigen peptide pRL1a generates a wide repertoire of CTL clones that differ in TCR V beta usage and in the intrapeptide epitope residues they recognize.

Animals↗
PubMed1997

Identification and characterization of functional residues in a Na+/H+ antiporter (NhaA) from Escherichia coli by random mutagenesis.

Forty-one mutants were isolated by means of random PCR mutagenesis of the Escherichia coli Na+/H+ antiporter (nhaA), which could not support the growth of a nhaAnhaB mutant (HIT delta AB-) on plates containing 0.15 M LiCl (pH 7.5) or 0.65 M NaCl (pH 8.0). Most of the mutants were sensitive to both NaCl and LiCl, or to LiCl alone. DNA sequencing revealed that twelve of the mutants had single amino acid substitutions. All the mutations, except for H225P, were of the conserved residues of NhaA homologues and located in the putative transmembrane helices. The Na+/H+ and Li+/H+ antiporter activities of the mutant NhaA were measured with everted membrane vesicles: eight of the mutants lost both antiporter activities completely under all pH conditions examined. Although both D133A and L138P retained low Li+/H+ antiporter activity, D133A lost Na+/H+ antiporter activity, while L138P retained normal Na+/H+ antiporter activity at pH 7.0 and 8.0. Interestingly, at pH 8.5, L138P no longer showed any Li+/H+ antiporter activity. H225P retained relatively high antiporter activities, although their pH dependence was altered. These observations supported the previous indication that His-225 is part of the pH sensor [Gerchman, Y. et al. (1993) Proc. Natl. Acad. Sci. USA 90, 1212-1216]. L73R exhibited about 20% each of the activities only at pH 8.0, and showed a similar pH dependence to H225P in both Na+/H+ and Li+/H+ antiport. Therefore, in addition to His-225, Leu-73, and/or its vicinity may also contribute to the pH sensing.

Amino Acid Sequence↗
PubMed1997

Physiological roles of endothelium-derived nitric oxide in the epigastric island flaps of rabbits.

Nitric oxide (NO), identified as the mediator of endothelium-dependent relaxation of vascular smooth muscle, is known to cause a number of inflammatory conditions, especially in ischemia/reperfusion injury. This experimental study, using a rabbit epigastric island flap, was designed to investigate whether skin flap ischemia followed by reperfusion-influenced serum NO and c-GMP concentrations in the flap. In addition, we also investigated the premedicated effects of the NO synthase inhibitor and heparin on serum NO and c-GMP concentrations in skin flap ischemia/reperfusion. Serum NO concentration after 15, 30, 45, and 60 minutes of ischemia followed by reperfusion significantly increased compared with that in nonischemic control and elevated flaps. On the contrary, serum NO concentration was suppressed in L-NAME or aminoguanidine pretreated animals with ischemic group. Administration of heparin increased the serum NO concentration in elevated flaps, but suppressed it in ischemic flaps followed by reperfusion. The changes in serum c-GMP and NO concentrations were related in all of the experimental groups. These results suggest that NO may be derived from vascular endothelial cells and dilate peripheral vessels in compensation for ischemia.

Animals↗
PubMed1997

Imaging assessment of the response of bone tumors to preoperative chemotherapy.

Assessment of the response of bone tumors to preoperative chemotherapy is of clinical importance. The authors determined the value of 3 imaging techniques (digital subtraction angiography, thallium scintigraphy, and dynamic magnetic resonance imaging) in guiding patient management by assessing the response of 17 bone sarcomas to preoperative chemotherapy compared with histologic evaluation of the resected specimens. Digital subtraction angiography showed a sensitivity of 87.5%, specificity of 57.1%, and accuracy of 73.3%. Thallium scintigraphy (sensitivity, 85.7%; specificity, 85.7%; accuracy, 85.7%) was superior to angiography in predicting tumor responses. The results of dynamic magnetic resonance imaging were analyzed on the basis of the value of slopes, which represents the percent increase in signal intensity per minute. The differences in slope before and after chemotherapy and the postchemotherapy slope values correlated with the histologic responses. The assessment by dynamic magnetic resonance imaging showed a sensitivity of 100%, specificity of 85.7%, and accuracy of 90.9%. Thallium scintigraphy and dynamic magnetic resonance imaging were considered noninvasive, reliable techniques that had about equal ability to assess the response of bone sarcomas to preoperative chemotherapy. Dynamic magnetic resonance imaging offers major advantages in the spatial resolution and can be more readily quantitated when compared with thallium scintigraphy.

Adolescent↗