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Biomedical subjects

S Ueda

Publications and source records attributed to S Ueda.

At least 343 records · Page 19Linked to original sources

Effect of cyclosporin A on isolated rabbit bladder and urethral smooth muscle.

BACKGROUND: The effects of Cyclosporin A (CsA) on vascular smooth muscle inducing hypertension have been established. However, there is no information available concerning the effects of CsA on urinary smooth muscle. Therefore, the effects of CsA were investigated on the bladder and urethral smooth muscle in rabbits. METHODS: CsA (10 mg/kg/day, intravenously) was given to rabbits for 14 days. Rabbit bladder and urethral smooth muscles were then isolated and evaluated using a muscle bath technique. RESULTS: 10(-10)-5 x 10(-6)M CsA alone had no direct effect on bladder or urethral smooth muscles. The Emax (maximum contractile response) and ED50 values for acetylcholine-induced contractions and the Emax (maximum relaxation response) for isoproterenol-induced relaxation in bladder smooth muscles were not significantly different between CsA-treated and control groups. The ED50 for isoproterenol-induced relaxation was significantly lower in the CsA group (P < 0.05). The Emax for phenylephrine- and clonidine-induced contractions in urethral smooth muscle was significantly higher in the CsA-treated group (P < 0.05). The ratio of the maximum response of the urethral smooth muscle to phenylephrine and clonidine in Ca+2-free solution to the normal solution in the CsA group (13.42% and 4.40%, respectively) was significantly higher than the maximal response ratios in the control group (6.34% and 3.00%, respectively; P < 0.05). CONCLUSIONS: CsA treatment augments the relaxation response of the bladder to isoproterenol and the contractile response of the urethra to phenylephrine and clonidine. In addition, CsA increases the filling of intracellular stores of releasable Ca+2, and also increases the permeability of Ca+2 in rabbit urethral smooth muscle. Thus, it is suggested that CsA may cause a urinary disturbance in patients treated with CsA via increased urethral resistance.

Acetylcholine↗

Subcapsular hematoma and hypertension following percutaneous needle biopsy of a transplanted kidney.

Percutaneous needle biopsy of a transplanted kidney is indicated as a diagnostic procedure in the evaluation of unexplained deteriorated renal function after renal transplantation. This procedure is not always without complications. We report a case of a subcapsular hematoma and hypertension following a percutaneous needle biopsy of a transplanted kidney. The cause of hypertension in this patient and the management of the subcapsular hematoma are discussed.

Adult↗

Latex agglutination test for canine parvovirus.

Canine parvovirus (CPV) was detected in faeces from dogs with diarrhoea by a specific slide agglutination test using latex particles coated with anti-CPV monoclonal antibody (LA-anti-CPV). The agglutination of LA-anti-CPV with CPV on a glass slide was evident macroscopically within 2 min. The sensitivity of the latex agglutination (LA) test was similar to that of the hemagglutination test. The LA test is available for the rapid diagnosis of CPV infection at an animal hospital.

Animals↗

Regulation of RNA polymerase sigma subunit synthesis in Escherichia coli: intracellular levels of four species of sigma subunit under various growth conditions.

By a quantitative Western immunoblot analysis, the intracellular levels of two principal sigma subunits, sigma 70 (sigma D, the rpoD gene product) and sigma 38 (sigma S, the rpoS gene product), and of two minor sigma subunits, sigma 54 (sigma N, the rpoN gene product) and sigma 28 (sigma F, the rpoF gene product), were determined in two Escherichia coli strains, W3110 and MC4100. The results indicated that the levels of sigma 54 and sigma 28 are maintained at 10 and 50%, respectively, of the level of sigma 70 in both strains growing at both exponential and stationary phases, but in agreement with the previous measurement for strain MC4100 (M. Jishage and A. Ishihama, J. Bacteriol. 177:6832-6835, 1995), the level of sigma 38 was undetectable at the exponential growth phase but increased at 30% of the level of sigma 70 at the stationary phase. Stress-coupled change in the intracellular level was observed for two sigma subunits: (i) the increase in sigma 38 level and the decrease in sigma 28 level upon exposure to heat shock at the exponential phase and (ii) the increase in sigma 38 level under high-osmolality conditions at both the exponential and stationary phases.

Bacterial Proteins↗

Molecular analysis of the poly(3-hydroxyalkanoate) synthase gene from a methylotrophic bacterium, Paracoccus denitrificans.

A 3.6-kb EcoRI-SalI fragment of Paracoccus denitrificans DNA hybridized with a DNA probe carrying the poly(3-hydroxyalkanoate) (PHA) synthase gene (phaC) of Alcaligenes eutrophus. Nucleotide sequence analysis of this region showed the presence of a 1,872-bp open reading frame (ORF), which corresponded to a polypeptide with a molecular weight of 69,537. Upstream of the ORF, a promoter-like sequence was found. Escherichia coli carrying the fusion gene between lacZ and the ORF accumulated a level of poly(3-hydroxybutyrate) that was as much as 20 wt% of the cell dry weight in the presence of beta-ketothiolase and acetoacetylcoenzyme A reductase genes of A. eutrophus. The ORF was designated phaCPd. A plasmid vector carrying the phaCPd'-'lacZ fusion gene downstream of the promoter-like sequence expressed beta-galactosidase activity in P. denitrificans. When a multicopy and broad-host-range vector carrying the ORF along with the promoter-like sequence was introduced into P. denitrificans, the PHA content in the cells increased by twofold compared with cells carrying only a vector sequence.

Acyltransferases↗

Sensitivity of volume-sensitive Cl- conductance in human epithelial cells to extracellular nucleotides.

The sensitivity to extracellular nucleotides of volume-sensitive Cl- channel activity was investigated by whole cell and single-channel patch-clamp recordings in a human small intestinal epithelial cell line (Intestine 407) during steady osmotic swelling. Adenine nucleotides added to the bathing solution suppressed whole cell volume-sensitive Cl- currents with the potency sequence of ATP > ADP > AMP. In contrast, extracellular adenosine 3',5'-cyclic monophosphate (cAMP) at over 0.1 mM increased volume-sensitive Cl- currents in the entire voltage range examined, whereas guanosine 3',5'-cyclic monophosphate was without effect. Neither the single-channel conductance nor the open probability was affected by extracellular cAMP. Extracellular ATP (at over 30 microM), in the Mg(2+)-free form, inhibited the whole cell volume-sensitive Cl- current, preferentially in the outward direction. By exposure to extracellular ATP, the single Cl- channel current became flickery at positive potentials. These results indicate that the volume-sensitive Cl- channel in the human epithelial cell is stimulated voltage independently by extracellular cAMP but blocked voltage dependently by the Mg(2+)-free form of extracellular ATP.

Adenosine Diphosphate↗

Irradiation effects on the metabolism of metastatic brain tumors: analysis by positron emission tomography and 1H-magnetic resonance spectroscopy.

To evaluate irradiation effects on the metabolism of metastatic brain tumors treated by Gamma Knife radiosurgery, positron emission tomography (PET) and 1H-magnetic resonance spectroscopy (MRS) studies were performed on five patients. The tumor origins were lung cancer in three patients and breast cancer in two. Treatment volume was 0.4-10.1 cm3 (mean: 5.5 cm3). The marginal dose to the tumor was 24-30 Gy (mean: 26.2 Gy). The follow-up period was 5-19 months (mean: 13.4 months). No patients had conventional whole-brain radiation therapy. 18F-fluoroboronophenylalanine (18FBPA) or 18F-fluorodeoxyglucose (18FDG) were used as tracers for the PET study. Using 1H-MRS, several metabolites were simultaneously measured in metastatic brain tumor and adjacent brain. In the PET study of the representative case, the uptake rate of 18FBPA that is actively transported to the tumor decreased markedly 15 days after radiosurgery and continued to decrease thereafter. In the 1H-MRS study, choline, which is characteristically high in metastatic brain tumors, also decreased over time. In two cases with suspected radiation injury, the enhanced region, which was decreased in size in early follow-up, enlarged progressively and was accompanied by edema. However, 18FBPA and 18FDG were not transported to the enhanced region. The peak of free lipid, which might show destruction of the cell membrane, was recognized in the enhanced region and adjacent brain in these cases. This study revealed that radiation effects on the metabolism of metastatic brain tumors occur at an early stage after radiosurgery and continue over several months. In particular, in the case of radiation injury, PET and 1H-MRS studies made it possible to distinguish between regrowth of the tumor and radiation injury.

Aged↗

Molecular anatomy of human chromosome 9: comparative mapping of the immunoglobulin processed pseudogene C epsilon 3 (IGHEP2) in primates.

Karyotypic homology in relation to human chromosome 9 (HSA 9) was studied through comparative mapping of the immunoglobulin-processed pseudogene C epsilon 3 (IGHEP2) in primates. IGHEP2, which has been mapped to 9p24.2 --> p24.1 in the human genome, was assigned to PTR 11q34 (common chimpanzee), PPA 11q34 (pygmy chimpanzee), PPY 13q16 (orangutan), HLA 8qter (white-handed gibbon), HAG 8qter (agile gibbon), and MFU 14q22 (Japanese macaque) by fluorescence in situ hybridization. To verify the breakpoints of presumed pericentric inversions on the ancestral great ape chromosomes, three DNA markers on HSA 9, cCI9-37 (9q22.1 --> q22.2), cCI9-135 (9q22.32 --> q22.33), and cCI9-208 (9p13.3 --> p13.2), were also assigned to PTR/PPA 11p11 (cCI9-37 and 135), PTR/PPA 11q22 (cCI9-208), PPY 13q22 (cCI9-37 and 135), and PPY 13q12 (cCI9-208). These data more clearly define the position of the breakpoints of pericentric inversions that occurred in the human-chimp ancestral and chimpanzee ancestral chromosomes and support the hypothesis of HSA 9 genesis previously derived from banding analyses of HSA 9 and its homologs.

Animals↗

The role of genetic polymorphisms of angiotensin-converting enzyme in the progression of renal diseases.

The renin-angiotensin system is likely to be important in the progression of renal diseases because of its effect on tissue hemodynamics and glomerular cell function. Recent evidence from small studies has suggested a possible role for the genetic determinants of angiotensin converting enzyme activity in the rate of progression of renal failure. We studied the effect of the insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme gene on the rate of renal function deterioration in 822 patients with a variety of renal diseases. We found that the slope of the reciprocal serum creatinine-versus-time plot was steeper in patients homozygous for the deletion allele (DD) compared with those homozygous for the insertion allele (II) (P = .015). When patients with similar renal function at presentation (creatinine < 200 mumol/L) were compared, II homozygotes had significantly improved renal survival (P = .039). Separate analyses of patients with glomerular diseases and tubulointerstitial diseases demonstrated an effect of this genotype in glomerular diseases only. These data provide further evidence of the possible role of the angiotensin-converting enzyme gene in the rate of progression of renal failure, although further studies are required to evaluate the role of this and other proposed candidate genes in renal diseases.

Adolescent↗

Suppression of enveloped virus production with a substance from silkworm faeces.

In the course of investigating effective biological active substances, we detected a substance in an extract of silkworm faeces that markedly suppresses viral production. The extract, prepared with hot phosphate-buffered saline and purified with ammonium sulfate precipitation, inhibited HVJ (Sendai virus), HSV (herpes simplex virus type-1), and HIV (human immunodeficiency virus type-1), but not poliovirus, suggesting that it is effective on enveloped virus production but not on non-enveloped ones. In the case of HVJ, indirect immunofluorescent staining using anti-HVJ antibody and Northern blotting analysis showed that, while viral adsorption and entry into the host cells were not affected, the synthesis of viral specific gene was inhibited by pretreatment of the virions with the extract. The extract affected more effectively aged virion, which losses membrane function as barrier and its envelope is leaky, than young virion that maintains barrier function. The active substance was partially purified by gel filtration after treatment of the extract with 1 N NaOH solution. From analysis with SDS-PAGE (SDS-polyacrylamide gel electrophoresis), protein bands were detected with molecular masses of about 25 kDa and near 14 kDa, while sugars were also detected with lectin blotting.

Animals↗

Calcitonin gene-related peptide inhibits human platelet aggregation.

Calcitonin gene-related peptide (CGRP) is a potent vasodilator in humans. CGRP receptors have also been found in various tissues other than blood vessels, such as the central nervous system, kidney, and heart. However, little is known about the effects of CGRP on human platelets. We investigated the effect of CGRP (human alpha type) on platelet aggregation in 21 healthy subjects (9 men and 12 women, mean age 54.6 years). CGRP inhibited platelet aggregation in vitro in 19 of the subjects (90.5%) in a dose-dependent manner with 50% inhibitory doses of 1.6 mumol/L and 1.1 mumol/L for aggregation induced by epinephrine and collagen, respectively. 125I-labeled CGRP bound specifically to intact platelets. The dissociation constant (Kd) was 61.9 +/- 17.7 pmol and the maximum number of binding sites (Bmax) was 5.4 +/- 3.9 pmol/10(9) platelets. The CGRP analogue (8-37)CGRP, but not calcitonin, inhibited the binding of 125I-CGRP to platelets. CGRP (5 mumol/L), but not (8-37)CGRP or calcitonin, increased the platelet cyclic AMP (cAMP) concentration by 31.7 +/- 3.6%. Thus, CGRP inhibits platelet aggregation via a specific receptor and by increasing the platelet cAMP concentration. CGRP may play a role in the modulation of platelet function in humans.

Adult↗

Mouse lethal activity of a HEp-2 vacuolation factor, cereulide, produced by Bacillus cereus isolated from vomiting-type food poisoning.

The HEp-2 vacuolation factor (or cereulide) produced by Bacillus cereus isolated from vomiting-type food poisoning, which is supposed to induce emesis, was found to give mouse and suncus lethality after intravenous and intraperitoneal administration. The emetic activity of the factor was also found to be resistant to heating at 121 degrees C for 15 min, exposure to pH 2 and 11, and to digestion with proteolytic enzymes such as pepsin and trypsin. These findings suggest that the cereulide produced by B. cereus is stable in the digestive tracts, induce emesis, and show lethal activity leading to cellular damage.

Animals↗

Cytopathic effect inhibition assay for canine interferon activity.

Conditions for cytopathic effect (CPE) inhibition assay of canine interferon (IFN) activity in Madin-Darby canine kidney (MDCK) cells and in canine tumor cell line A72 was investigated using the New Jersey strain of vesicular stomatitis virus (VSV). The culture supernatant from canine splenocytes stimulated with ultraviolet-irradiated Newcastle disease viruses was used as reference IFN. MDCK cells were resistant for growth of VSV when the cells were confluent. Full CPE was observed only in a sparsely growing culture. Canine IFN activity could be assayed on less than 10(4) MDCK cells/well of a 96-well microplate, and more than 10(5) TCID50/ml of VSV was required. In A72 cells, VSV growth was not as dependent on cell density as in MDCK cells, requiring 10(3) TCID50/ml of VSV. MDCK-VSV system showed a higher IFN sensitivity than A72-VSV, whereas reproducibility was higher for the latter than the former. Based on these findings, A72-VSV system for canine IFN assay is recommended for practical use due to its easy handling characteristics.

Animals↗

Protective immunity induced with the antigens fractionated by affinity chromatography from the second generation schizonts of Leucocytozoon caulleryi.

A crude extract of the 2nd generation schizonts (2GS) of Leucocytozoon caulleryi and chicken serum containing serum-soluble antigens were applied to an immunoaffinity column using immunoglobulin of chickens recovered from L. caulleryi infections. The fractionated antibody-bound proteins showed several bands between 70 kDa and 35 kDa by the electrophoretic analysis. By immunization of chickens, the fractionated proteins conferred the protection at varying degrees comparable to that conferred by the unfractionated original materials. These results indicated that antiserum against 2GS can recognize the proteins involved in protective immunity.

Animals↗

Mistyping of the human angiotensin-converting enzyme gene polymorphism: frequency, causes and possible methods to avoid errors in typing.

A polymorphism of the gene encoding the human angiotensin I-converting enzyme (ACE), which is defined by an insertion/deletion polymorphism in intron 16, has been identified as a candidate genetic locus in the development of cardiovascular and renal disease. We have demonstrated that the accuracy of ACE genotyping is critically dependent on the strategy of the PCR used in typing. Of 1238 individuals genotyped by a standard method, 335 were typed as DD, 645 as DI and 258 as II. However, when DD individuals were retyped using modified methods (including either 5% dimethyl sulphoxide, or a 'hot start') 35 of the original 335 samples (10.5%) were retyped as DI. In approximately half of these mistyped samples, PCR amplification was assessed as inefficient by the absence of a third faint heteroduplex band in a control ID sample: when the assay was repeated without any modifications, the mistyped samples were correctly genotyped. In the remainder, mistyping persisted. In these cases, the use of a third 'nested' PCR primer specific for the I allele was required for successful genotyping, providing a more reliable strategy without the need for further modification to the PCR technique. Our results suggest that the triple primer approach is the method of choice for accurate ACE genotyping.

Dimethyl Sulfoxide↗

Traumatic acute subdural hematoma localized on the superior surface of the tentorium cerebelli--two case reports.

An 8-year-old boy, who fell downstairs and struck his head, and a 62-year-old female, who hit her head in the automobile accident, presented with unusual traumatic acute subdural hematoma localized on the superior surface of the tentorium cerebelli. Magnetic resonance imaging was useful for determination of the anatomical location of the hematoma, and confirmation of absence of significant parenchymal contusion. Injury of the variant bridging vein possibly caused subdural hematoma over the tentorium.

Child↗

Effectiveness of revascularization surgery evaluated by proton magnetic resonance spectroscopy and single photon emission computed tomography.

Proton magnetic resonance spectroscopy (MRS) and single photon emission computed tomography (SPECT) were used to evaluate chronic ischemic regions in 26 stroke patients before and 1, 3, and 6 months after revascularization surgery. The volume of interest for proton MRS was placed in an area including part of the frontal and temporal opercula, insular cortex, and basal ganglia. Twenty healthy volunteers served as controls for proton MRS. Patients were divided into three groups according to the preoperative proton MRS. Group A (n = 12) had significantly lower N-acetylaspartate/choline (NAA/Cho) and N-acetylaspartate/creatine (NAA/Cr) ratios on the operative side compared to those on the contralateral side, and also lower than those in normal subjects. In seven patients in Group A, postoperative serial proton MRS demonstrated no recovery of these ratios on the operative side. However, proton MRS of the other five patients indicated gradual improvement in these ratios on the operative side at 3 to 6 months after surgery, and SPECT indicated an increase in cerebral blood flow on the operative side in four of these five patients. In Group B (n = 9), proton MRS and SPECT showed no laterality before revascularization and no remarkable change during the postoperative course. In Group C (n = 5), NAA/Cho or NAA/Cr decreased on the contralateral side preoperatively. Two patients showed fluctuating values of NAA/Cho or NAA/Cr during the postoperative period. Serial proton MRS and SPECT studies may be useful for the evaluation of revascularization surgery on ischemic regions. The efficacy of revascularization surgery on the metabolism may appear gradually within 3-6 months.

Adolescent↗