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Biomedical subjects

H Ueda

Publications and source records attributed to H Ueda.

At least 397 records · Page 22Linked to original sources

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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

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↗

HLA associations in otosclerosis in Japanese patients.

Otosclerosis is a disease of the otic capsule that is caused by abnormal resorption and redeposition of bony tissue. Sixty-two unrelated Japanese patients exhibiting clinical otosclerosis were typed for HLA-A, -B, -C antigens. Twenty-one of the patients were also typed for DR antigens. The frequency of HLA-Aw33 was significantly higher in otosclerosis patients than in the control group (24.2% vs 9.5%). This finding suggests that the presence of HLA-Aw33 antigens may be related to an increased susceptibility to otosclerosis or to its clinical outcome.

Adolescent↗

Dynamic structure of glomerular capillary loop as revealed by an in vivo cryotechnique.

Morphological studies using immersion or perfusion fixation methods do not reveal the ultrastructure of functioning kidneys with normal circulation. A simple apparatus was developed for freezing the kidneys in vivo without stopping the blood supply, and the ultrastructure of the glomerular capillary loops was examined under different haemodynamic conditions. Mouse kidneys were frozen under normal blood flow conditions; others were frozen in the same way after ligation of the abdominal aorta at a point caudal to the renal arteries. They were then processed for the freeze-substitution or deep-etching method. Good ultrastructural preservation was obtained within about 5 microM depth from the frozen tissue surface. Functioning glomeruli with normal blood flow possessed open capillary lumens, different shapes of foot processes and atypical basement membranes with low density. Moreover, heterogeneity in width between foot processes was identified on the replica membranes. Under the acute conditions used to increase blood supply into the kidneys, the spaces between the flat foot processes became more widely dilated and the basement membrane was seen to be three-layered. The ultrastructure of glomeruli in functioning kidneys has been demonstrated for the first time by this "in vivo cryotechnique."

Animals↗

Sensitization interval and administration method alter the effect of 15-deoxyspergualin on heart transplantation in sensitized recipient rats.

We evaluated the effect of 15-deoxyspergualin (DSG) on accelerated rejection. Brown Norway rats (BN) served as organ donors and Lewis rats (LEW) as recipients. In an accelerated rejection model, after a LEW rat was sensitized with BN skin, a BN heart was transplanted. Various intervals between sensitization and heart transplantation were examined. The heart allografts in sensitized recipients were rejected earlier than those in unmodified recipients regardless of the sensitization interval. DSG (2.5 mg/kg per day), given to the recipients during the sensitization phase, significantly prolonged graft survival compared with the untreated hosts when the sensitization interval was short. When the recipients were treated with DSG after heart transplantation, heart graft survival was significantly prolonged regardless of the sensitization interval. Flow cytometric analysis and complement-dependent cytotoxicity tests revealed that DSG suppressed antidonor antibody formation and the postoperative administration of DSG significantly decreased the proliferation of B cells when the sensitization interval was short and the proliferation of class II antigen-positive cells when the sensitization interval was long.

Animals↗

Association of Trp64Arg mutation of the beta3-adrenergic-receptor with NIDDM and body weight gain.

A possible pathogenic mutation in the beta 3-adrenergic-receptor gene (Trp64Arg) has been reported to be associated with an earlier age of onset of non-insulin-dependent diabetes mellitus (NIDDM) and clinical features of the insulin resistance syndrome in Pima Indian, Finnish and French subjects. Since marked heterogeneity has been reported in the association of mutations of candidate genes with NIDDM between Japanese and other ethnic groups, we investigated the association of Trp64Arg with NIDDM in Japanese subjects. The allele frequency of the mutation (Arg) was slightly, but not significantly, higher in NIDDM than in control subjects (70 out of 342 alleles [20.5%] vs 40 out of 248 [16.1%], respectively, p > 0.2). When our data were combined with those of Pima Indian and Finnish subjects, however, the Arg/Arg genotype was significantly associated with NIDDM as compared with the other two genotypes (p < 0.005, relative risk [RR] 2.13, 95% confidence interval [CI] 1.28-3.55). The Arg allele was also associated with NIDDM (p < 0.05, RR 1.27, 95% CI 1.06-1.52). Japanese subjects homozygous for the mutation had a significantly higher body mass index (mean +/- SD: 25.5 +/- 3.9 kg/m2) than heterozygotes (22.6 +/- 4.1, p < 0.05) and normal homozygotes (22.8 +/- 3.8, p < 0.05). NIDDM patients homozygous for the mutation tended to have an earlier age of onset of NIDDM than those with other genotypes. These data suggest that the Trp64Arg mutation not only contributes to weight gain and age-at-onset of NIDDM but is also associated with susceptibility to NIDDM.

Age of Onset↗

Clothing microclimate temperatures during thermal comfort in boys, young and older men.

To examine the effects of age-related differences in thermoregulatory function on the clothing microclimate temperature (Tm) and Tm fluctuations while maintaining thermal comfort in daily life, 5 boys (group B, 10-11 years), 5 young men (group Y, 20-21 years) and 5 older men (group O, 60-65 years) volunteered to take part in this study. The subjects were asked to maintain thermal comfort as closely as possible in their daily lives. Tm (temperatures between the skin surface and the innermost garment) at four sites (chest, back, upper arm, and thigh), skin temperature on the chest (Tchest) and ambient temperature (Ta) were measured over a period of 8-12 h from morning to evening on one day in each of the seasons, spring, summer, autumn, and winter. Records of ability to maintain thermal comfort and of adjustment of their clothes were kept by each subject. Ta during periods of thermal comfort did not differ among the groups in any of the seasons. In group Y, Tm was significantly lower at the thigh than at the other sites in spring, autumn, and winter (P < 0.05) and fluctuations (CV) of Tm were significantly larger at the thigh than at other sites in autumn and winter (P < 0.05). Similar tendencies were observed for Tm and CV of Tm in group B. However, Tm and CV of Tm in group O did not differ by site except for the autumn Tm. Group O had a smaller CV at the thigh in winter (P < 0.05), compared to groups B and Y, suggesting a smaller regional difference in Tm fluctuation in group O. Group O adjusted their clothes even on the lower limbs (together with upper body) in order to maintain thermal comfort in accordance with changes in Ta, while groups B and Y did so only on their upper bodies. These results suggest that compared to boys and young men, lower thermoregulatory function in older men may affect Tm and CV of Tm as a result of clothing on lower limbs being adjusted differently in order to maintain thermal comfort.

Adult↗