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Biomedical subjects

H Ueda

Publications and source records attributed to H Ueda.

At least 145 records · Page 8Linked to original sources
PubMed1997

Expression of endothelial nitric oxide synthase in human eccrine clear cells.

Nitric oxide is generated from L-arginine by nitric oxide synthase (NOS), which has at least three isoforms; endothelial-type NOS (eNOS) and brain-type NOS (bNOS) are constitutive enzymes, and inducible-type NOS (iNOS) is expressed after stimulation. Studies by the avidin-biotin immunocomplex method, revealed eNOS immunoreactivity exclusively in the human eccrine clear cells. No eNOS immunoreactivity was observed in the eccrine dark cells or myoepithelial cells. No staining of iNOS or bNOS was observed in the eccrine gland. These findings indicate that NO plays a physiological part in the production and/or excretion of sweat in the human skin eccrine gland.

Adult↗
PubMed1997

Immunocytochemical study of dystrophin localization in cone cells of mouse retinas.

PURPOSE: Previously, the authors reported that dystrophin was observed under the rod cell membranes in rat retinas. However, it was not determined whether dystrophin is located in cone cells. In the current study, the authors clarify dystrophin localization in cone cells of mouse retinas. METHODS: Immunoblotting, confocal laser scanning microscopy, and immunoelectron microscopy were used to investigate retinal dystrophin with a monoclonal antibody raised against the human dystrophin C-terminus. RESULTS: Immunoblotting analysis showed some immunoreactive bands from retinal extracts. Confocal images indicated two different immunostaining patterns: One was a tiny dot, and the other was a larger, aggregated dot. Immunoelectron microscopy revealed that retinal dystrophin was localized in cone cells as well as in rod cells. CONCLUSIONS: Retinal dystrophin is a common component of cone and rod cells and probably is related to the physiological function of photoreceptor cells.

Animals↗
PubMed1997

Neuroprotective effect of D-(E)-2-amino-4-methyl-5-phosphono-3-pentenoic acid in the gerbil model of transient global cerebral ischemia.

Effect of the new competitive N-methyl-D-aspartate (NMDA) antagonist D-(E)-2-amino-4-methyl-5-phosphono-3-pentenoic acid (CAS 137424-81-8, CGP 40116) was examined in a mongolian gerbil model of global cerebral ischemia. Effect of CGP 40116 was compared to that of another competitive NMDA antagonist, (+/-)-cis-4-phosphonomethyl-piperadine-2-carboxylic acid (CAS 110347-85-8, CGS 19755) under the same conditions. Drugs were administered intraperitoneally 30 min before bilateral carotid artery occlusion. At 4 days after the ischemia, locomotor activity was significantly higher in ischemic control mongolian gerbils in comparison with sham-operated mongolian gerbils. CGP 40116 at the dose of 10 mg/kg and CGS 19755 at the doses of 10 and 30 mg/kg significantly suppressed the increase of the motility. Seven days after ischemia, ischemic control group was still hyperactive compared to sham-operated group. CGP 40116 at the dose of 10 mg/kg and CGS 19755 at the dose of 30 mg/kg significantly reversed it. The number of survived neurons of ischemic control group was significantly less than that of sham-operated group at 7 days after ischemia. CGP 40116 at the dose of 10 mg/kg and CGS 19755 at the dose of 30 mg/kg significantly increased the number of survived neurons. It is concluded that CGP 40116 is more potent for amelioration of global cerebral ischemic damage than CGS 19755.

2-Amino-5-phosphonovalerate↗
PubMed1997

Effect of D-(E)-2-amino-4-methyl-5-phosphono-3-pentenoic acid on focal cerebral ischemia in cat.

The effect of the new competitive N-methyl-D-aspartate (NMDA) antagonist D-(E)-2-amino-4-methyl-5-phosphono-3-pentenoic acid (CAS 137424-81-8, CGP 40116) was examined in a cat model of focal cerebral ischemia. Effect of CGP 40116 was compared to that of another competitive NMDA antagonist, (+/-)-cis-4-phosphonomethyl-piperadine-2-carboxylic acid (CAS 110347-85-8, CGS 19755) under the same conditions. Drugs were administered intravenously 30 min before left middle cerebral artery (MCA) occlusion. After MCA occlusion for 8 h, infarction spreaded widely among caudate nucleus prepyriform cortex, amygdala and temporal lobe cortex in the ischemic hemisphere. CGP 40116 at the dose of 10 mg/kg and CGS 19755 at the dose of 30 mg/kg significantly decreased the infarcted area. CGP 40116 was effective in the frontal and central brain, although CGS 19755 showed neuroprotective effect in almost all sites. Thus, the compounds are potent neuroprotectants in focal ischemia.

2-Amino-5-phosphonovalerate↗
PubMed1997

[Effect on 5'-deoxy-5-fluorouridine (5'-DFUR) of pyrimidine nucleoside phosphorylase (PyNPase), matrix metalloprotease and serum IAP values. Hokuriku Colorectal Cancer Chemotherapy Study Group].

PyNPase activity, MMPs activity and serum IAP values were measured in tumor tissues from colorectal cancer patients who had been divided into two groups, one given preoperative 5'-DFUR and the controls. PyNPase activity of the preoperative administration group was approximately equivalent to that of the controls. In the control group, correlations were assessed between PyNPase activity and activities of MMP1 and MMP3. To assess the effect of 5'-DFUR on the activity of MMPs, we divided patients into two groups, a high and a low PyNPase activity group. Although there was no correlation with MMPs activity of the preoperative administration group and the control group in the low PyNPase activity group, the activities of MMP1 and MMP9 of the control group were significantly higher in the high PyNPase activity group. Moreover, the serum IAP value of the administration group was significantly lower than that of the control group. These results indicated that PyNPase activity was thus suggested to be somehow related to MMPs activity and serum IAP values.

Adult↗
PubMed1997

[Ureteral cancer producing carbohydrate antigen 19-9: report of a case].

A case of a CA19-9-producing ureteral cancer is reported. A 58-year-old man presented with gross hematuria. Retrograde pyelography showed an irregular filling defect in the right ureter. The serum CA19-9 level was 932 U/ml (normal < 37). Right total nephroureterectomy was performed. The histological diagnosis was grade 3 transitional cell carcinoma. Immunohistochemical analysis showed CA19-9 to be expressed not only in the cancer cells but also in the normal transitional cell epithelium of the renal pelvis.

Biomarkers, Tumor↗
PubMed1997

Hemodynamic effects of warm bathing in a Hubbard tank and exercise loading in patients after myocardial infarction.

Hemodynamic parameters were measured during bathing and exercise testing in 43 patients with myocardial infarction (mean age: 60.2 years) to investigate the predictive parameters to determine when patients could safely resume bathing. Patients took a fresh water bath at 42 degrees C in the supine position for 5 min in a Hubbard tank. Group A showed an elevation of pulmonary capillary wedge pressure (PCWP) during bathing of 10 mmHg or more (23 patients, mean age: 61.7 years) and group B showed an elevation of less than 10 mmHg (20 patients, mean age: 60.5 years). Continuous multistep exercise tests were performed with a bicycle ergometer in the supine position, and hemodynamic parameters were measured at up to 50 W for 3 min on the day before the warm bathing test. There were no significant differences in the changes of arterial pressure and heart rate between the two groups. The PCWP at 3 min with a load of 50 W was significantly higher in group A (26.9 +/- 9.0 mmHg) than in group B (16.7 +/- 9.1 mmHg, p < 0.01). The stroke index (SI) during exercise testing was significantly lower in group A than in group B. The difference in the stroke index from baseline values (delta SI) at 3 min with a load of 50 W was significantly lower in group A (3.5 +/- 5.5 ml/m2/beat) than in group B (10.6 +/- 7.0 ml/m2/beat, p < 0.01). Similarly, delta CI and delta oxygen pulse during testing were significantly lower in group A than in group B. The physical work capacity and ejection fraction of the left ventricle of group A were significantly lower than those of group B, whereas the left ventricular end-diastolic pressure was higher in group A than in group B. CI, delta CI, SI, delta SI, METs, oxygen pulse, and delta oxygen pulse were examined by regression analysis and multivariate analysis to predict a significant elevation of delta PCWP during bathing. delta SI (p = 0.0032), delta CI (p = 0.0094), delta SI + METs (p = 0.0051), delta CI + METs (p = 0.0061), delta CI + delta SI (p = 0.0084), and delta CI + delta SI + METs (p = 0.0093) showed the highest correlations with delta PCWP. These findings suggest that changes in delta CI, delta SI, and METs are good predictive parameters for determining when patients may safely resume bathing. We suggest that patients with myocardial infarction, reduced cardiac function and a physical work capacity of approximately 4.0 METs, delta SI: 5 ml/m2/beat and delta CI: 2.4 l/min/m2 resume bathing only after careful consideration.

Adult↗
PubMed1996

Similarity of in vivo somatic mutations at an autosomal adenine phosphoribosyltransferase locus between T- and B-cells in human peripheral blood.

In vivo somatic mutations have been detected at several human loci by using clonal cultures of peripheral blood T-cells. It has not been fully understood whether or not the somatic mutations in T-cells are similar to those of other cell types. To address this issue, we cloned, from human peripheral blood, T- and B-cells with mutations at an autosomal adenine phosphoribosyltransferase (APRT) locus. For the efficient detection of somatic mutations at the APRT locus, a blood sample from a human individual heterozygous for germline APRT deficiency was used. T- and B-cells deficient in APRT enzyme activity were cloned from peripheral blood mononuclear cells using a selecting agent, 2,6-diaminopurine. The APRT-deficient mutant frequencies were on the order of 10(-4) in both T- and B-cells. The single-strand conformation polymorphism analysis of the APRT DNA of mutant B-cell clones suggested that the molecular mechanisms leading to the APRT deficiency in B-cells were similar to those in T-cells. Our observations suggest that both the frequency and the mode of in vivo somatic mutations occurring spontaneously at general autosomal loci in B-cells are similar to those in T-cells.

Adenine Phosphoribosyltransferase↗
PubMed1996

Efficient gene transfer to EGF receptor overexpressing cancer cells by means of EGF-labeled cationic liposomes.

Epidermal growth factor (EGF)-labeled cationic liposomes (EGF-liposomes) were prepared for efficient gene transfer vector to EGF receptor expressing cells. Transfection activity of EGF-liposomes associating plasmid PGV-C which encodes luciferase showed a 2-fold increase in EGF receptor expressing cells, HEC-1-A, compared to that of EGF-non-labeled liposomes (N-liposomes). In EGF receptor deficient HRA cells, however, both EGF- and N-liposomes exhibited low transfection efficiency and no difference was observed between them. Furthermore, by the addition of anti EGF receptor antibody, transfection efficiency of EGF-liposomes was suppressed, suggesting EGF receptor-mediated endocytosis of EGF-liposomes. Transfection activity of EGF-liposomes was strongly dependent on the concentrations of fusogenic lipid, dioleoylphospha-tidylethanolamine in liposomes. By X-gal staining 6-8% of GCH-1(m) cells which also had EGF receptor expressed beta-galactosidase activity following the transfection by EGF-liposomes associating pSV-beta-galactosidase. These findings indicate that EGF-liposomes could be a preferable vector for EGF-receptor expressing cells.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1996

Analysis of the mRNA cap-binding ability of human eukaryotic initiation factor-4E by use of recombinant wild-type and mutant forms.

In order to identify the amino acid residues necessary for the selective recognition of the mRNA cap structure by human eukaryotic initiation factor-4E (eIF-4E), which plays a central role in the first step of mRNA translation, we prepared recombinant wild-type and fourteen mutant forms and compared their cap-binding abilities by affinity chromatography. By the direct expression of a synthetic gene encoding human eIF-4E as the soluble form in Escherichia coli and the application on a 7-methylguanosine-5'-triphosphate-Sepharose 4B cap affinity column, pure recombinant eIF-4E was prepared; the optimum pH for the binding of the mRNA cap was 7.5. Among the amino acid residues conserved among various eIF-4E species, each of 14 functional residues was replaced with a nonpolar amino acid (alanine or leucine). All mutant eIF-4E genes, which were constructed by site-directed mutagenesis, were expressed in the same way as the wild type, and their cap-binding abilities were compared with that of the wild type. Consequently, all eight tryptophan residues. Glu103, and two histidine residues at positions 37 and 200 in human recombinant eIF-4E were suggested to be important for the recognition of the mRNA cap structure through direct interaction and/or indirect contributions. Indirect contributions included the construction of the overall protein structure, especially the cap-binding pocket.

Amino Acid Sequence↗
PubMed1996

Inositol 1,4,5-trisphosphate-gated calcium transport through plasma membranes in nerve terminals.

We developed new biochemical approaches to demonstrate the presence of inositol 1,4,5-triphosphate (InsP3)-gated calcium channels in presynaptic plasma membranes (SPM) and their involvement in the presynaptic receptor-mediated Ca2+ influx into nerve terminals. In perfusion experiments using SPM vesicles preloaded with 45Ca2+, InsP3 elicited the release of 45CA2+ into perfusates in a saturable manner. The InsP3- evoked 45Ca2+ release from resealed SPM vesicles was more potent than that from resealed vesicles using any other subcellular fractions. Here we also report the involvement of InsP3-gated mechanisms in the presynaptic receptor-mediated Ca2+ influx into synaptosomes (nerve terminals) by use of such resealed vesicles reconstituted with purified Gi1.

Adenosine Triphosphate↗
PubMed1996

Inhibition of tumor growth by direct intratumoral gene transfer of herpes simplex virus thymidine kinase gene with DNA-liposome complexes.

To establish the expression of the herpes simplex virus thymidine kinase (HSV-TK) gene in tumor cells, we analyzed the promoter function of the SV40 promoter and the nucleotide sequence (CACGTG) to which Myc-Max heterodimers (Myc/Max) were capable of binding in four kinds of cell lines: COLO320 DM, A-431, KF, and Nakajima. When luciferase reporter plasmid under the control of SV40 promoter was transfected into tumor cells in vitro, a high level of luciferase activity was observed in all kinds of cell lines. However, by transfection of the luciferase gene promoted by the Myc/Max binding sequence, accelerated luciferase expression was observed in COLO320 DM and A-431 cells with high expression of c-myc, but not in KF and Nakajima cells, which showed low expression of c-myc. The repeated transfection of the liposome-conjugated HSV-TK gene regulated by the SV40 promoter and cultivation in 100 micrograms/ml of aciclovir for 5 days in vitro demonstrated growth inhibition for all four kinds of cell lines. However, cell toxicity was observed only in COLO320 DM and A-431 cells when the HSV-TK gene promoted by the Myc/Max binding sequence was introduced. (The survival rate to 100 micrograms/ml of aciclovir concentration in COLO320 DM, A-431, KF, and Nakajima cells was 59%, 53%, 74%, and 79%, respectively.) In vivo direct injection of the liposome-conjugated HSV-TK gene regulated by the SV40 promoter into established tumors and aciclovir administration for 10 days into the mice resulted in significant tumor volume reduction in three tested tumor cells. However, injection of the HSV-TK gene promoted by the Myc/Max binding sequence and aciclovir administration into mice could achieve significant tumor regression only in COLO320 DM and A-431 cells. These results suggest that gene therapy using the HSV-TK gene promoted by the Myc/Max binding sequence can be an attractive approach for treatment against tumor cells expressing high levels of c-myc.

Acyclovir↗
PubMed1996

Regulation of transforming growth factor-beta 1 expression by the hepatitis B virus (HBV) X transactivator. Role in HBV pathogenesis.

TGF-beta 1 has been implicated in the pathogenesis of liver disease. The high frequency of detection of the hepatitis B virus X (HBx) antigen in liver cells from patients with chronic hepatitis, cirrhosis, and liver cancer suggested that expression of HBx and TGF-beta 1 may be associated. To test this possibility, we examined the expression of TGF-beta 1 in the liver of transgenic mice expressing the HBx gene. We show that the patterns of expression of TGF-beta 1 and Hbx protein are similar in these mice and that HBx activates transcription of the TGF-beta 1 gene in transfected hepatoma cells. The cis-acting element within the TGF-beta 1 gene that is responsive to regulation by Hbx is the binding site for the Egr family of transcription factors. We further show that the Egr-1 protein associates with the HBx protein, allowing HBx to participate in the transcriptional regulation of immediate-early genes. Our results suggest that expression of Hbx might induce expression of TGF-beta 1 in the early stages of infection and raise the possibility that TGF-beta 1 may play a role in hepatitis B virus pathogenesis.

Amino Acid Sequence↗
PubMed1996

Stapes fixation associated with symphalangia.

We describe a 10-year-old boy and his 9-year-old sister, both of whom had bilateral hearing loss associated with ankylosis of the proximal interphalangeal joints (symphalangia). They had no other abnormal findings except hearing loss and ankylosis of the joints of some fingers and toes on systemic examination. The cause of conductive hearing loss in both cases was bony fusion between the stapes and the bone of the oval window niche. There were no other anatomical abnormalities in the middle or the external ear. The patients' hearing improved markedly after stapes surgery. Histopathologic examination of the stapes revealed an abnormal zone of ossification near the anterior annular ligament and calcification in the annular ligament, which seemed to be the cause of the stapes fixation.

Child↗
PubMed1996

Cytokines involving gp130 in signal transduction suppressed growth of a mouse hybridoma cell line and enhanced its antibody production.

Three cytokines, interleukin 6 (IL-6), leukaemia inhibitory factor (LIF), and oncostatin M (OSM), that bind to composite receptors including a common signal transducer gp130 suppressed proliferation of a mouse B-cell hybridoma cell line 2E3-O cultured in serum-free medium, while they enhanced antibody production of the cells. The specific growth rate of the cells reduced from 1.0/day for control to 0.6/day for the cultures supplemented with IL-6, LIF, or OSM at 1, 4, or 2 ng/ml, respectively. The antibody productivity increased five-fold when the cells were cultured with IL-6, LIF, or OSM at 1, 25, or 20 ng/ml, respectively. Transforming growth factor beta1(TGF-beta1) similarly suppressed growth of the cells at the concentration of 5 ng/ml, while it did not enhance the antibody production. Cell cycle analysis revealed that IL-6 induced the cells to be arrested at G1 phase of the cell cycle more intensively than TGF-beta1, indicating that IL-6 and TGF-beta1 suppressed the growth through mutually different mechanisms. As a whole, this work suggests that gp130, which is commonly involved in each receptor for IL-6, LIF, and OSM, transduces signals for suppressing proliferation and possibly for enhancing antibody production in the hybridoma cells.

Animals↗
PubMed1996

The significant effect of HLA-DRB1 matching on acute rejection in kidney transplants.

The object of the present study was to confirm the HLA-DRB1 matching effect on rejection crisis, its severity, and kidney graft survival based on genotyping. Ninety-four renal allografts were included in this study. DNA typing of HLA-DRB1 was performed by the polymerase chain reaction sequence-specific oligonucleotide method. The incidence of acute rejection within 6 months following transplantation, the frequency of OKT3 administration for steroid-resistant rejection, histopathological findings, and graft survival rate were compared between the DRB1-matched (n = 23) and DRB1-mismatched (n = 71) groups. Four acute rejections occurred in the DRB1-matched group (incidence; 17%) and 40 in the DRB1-mismatched group (56%). In the DRB1-matched group, the incidence of acute rejection was significantly less frequent than that of the DRB1-mismatched group (P < 0.005). In the DRB1-matched group, only one patient received OKT3 administration (4%), in contrast to 16 of 71 patients in the DRB1-mismatched group (23%). The use of OKT3 was significantly less frequent in the DRB1-matched group (P < 0.05). Histopathological findings from biopsy specimens showed no constant distribution of pathological grades of acute rejection according to DRB1 matching in the present study. The graft survival rate in the two groups did not differ significantly, but the graft survival rate in the DRB1-mismatched group had a tendency to decrease as the grafts survived longer. In conclusion, the results of the present study confirm that HLA-DRB1 matching has marked beneficial effects on kidney transplants through reduction of the acute rejection rate and decrease of the severity of rejection, and suggest that improvement of graft survival will be obtained through kidney allocation to a DRB1-matched recipient.

Acute Disease↗