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Biomedical subjects

S Uchida

Publications and source records attributed to S Uchida.

At least 199 records · Page 11Linked to original sources

Apafant (a PAF receptor antagonist) suppresses the early and late airway responses in guinea pigs: a comparison with antiasthmatic drugs.

We studied the effects of apafant (WEB 2086 BS), a specific and potent platelet activating factor (PAF) receptor antagonist, on the early and late airway responses in conscious and actively sensitized guinea pigs. An increase in airway resistance (Rs) was seen 1 min after the inhaled antigen challenge (early airway response), followed by another increase in Rs which peaked between 4 and 8 h after the provocation (late airway response). Oral administration of apafant as well as theophylline inhibited both early and late airway responses. Ozagrel, an inhibitor of thromboxane A2 synthetase, salbutamol, a beta2-adrenoceptor agonist, and dexamethasone significantly inhibited either the early or the late airway response only. Disodium cromoglycate inhibited neither the early nor the late airway response. The results showed that apafant inhibited both the early and late airway responses in sensitized guinea pigs and its effect was comparable or superior to that of anti-asthmatic drugs used clinically.

Administration, Inhalation↗

Regulation of aquaporin-2 gene transcription by GATA-3. off.

To evaluate the functional role of GATA motifs in the 5'-flanking region of a kidney-specific AQP-2 water channel gene, we sought to isolate a GATA factor(s) expressed in collecting ducts and determined the role on the AQP-2 promoter. Two cDNAs encoding GATA factors were isolated from rat kidney, whose sequences were highly homologous with human GATA-2 and -3. Reverse-transcription PCR using dissected nephron segments revealed that rat GATA-3 but not GATA-2 was expressed in collecting ducts, thus indicating that GATA-3 could interact with GATA motifs in the AQP-2 promoter. Transactivation experiments utilizing the rat GATA-3 expression vector indicated that rat GATA-3 increased the AQP-2 promoter activity about fourfold. These results indicated that GATA motifs in the 5'-flanking region of the hAQP-2 gene were functional cis-elements and that GATA-3 in collecting ducts may be one of the important regulators of AQP-2 expression in vivo.

Animals↗

Characterization of T-cell clones derived from peripheral blood lymphocytes of a patient with transfusion-associated graft-versus-host disease: Fas-mediated killing by CD4+ and CD8+ cytotoxic T-cell clones and tumor necrosis factor beta production by CD4+ T-cell clones.

Transfusion-associated graft-versus-host disease (TA-GVHD) is one of the most serious adverse effects of blood transfusion. It is generally thought to be caused by the infused lymphocytes. Donor-derived cytotoxic T lymphocytes (CTLs) directed against the recipient's HLAs, which have escaped the recipient's immune system and are proliferating, are considered to attack recipient organs and tissues. Despite the seriousness of the disease, the precise mechanism of its development remains unclear and no definitive treatment has been developed. With the aim of developing an effective treatment, we established and characterized T-cell clones from peripheral blood lymphocytes (PBLs) of a TA-GVHD patient. Three types of clones were established. Type I clones were CD8+ and specifically lyse cells that express HLA B52. Type II clones were CD4+, specifically lysed cells that express HLA DR15, and proliferated in response to stimulation with cells that express DR15. Type III clones were also CD4+, showed no cytotoxic activity toward any HLA-expressing cells, and proliferated in response to stimulation with cells that express DR15. Furthermore, we found that the Fas/Fas-ligand (Fas-L) system is involved in the cytotoxicity of the type I and II clones and that the type III clones produce and secrete a large amount of tumor necrosis factor beta (TNFbeta) after antigen stimulation. Based on our results, these three types of clones can be classified into two categories: those that have the ability to induce GVHD directly by cytolysis and that show no cytotoxic activity and those that have the ability to cause GVHD indirectly through secretion of cytotoxic lymphokines.

Aged↗

Effect of regional myocardial perfusion abnormalities on regional myocardial early diastolic function in patients with hypertrophic cardiomyopathy.

Nonuniform hypertrophy of the left ventricle is an important factor in regional diastolic dysfunction in patients with hypertrophic cardiomyopathy (HCM). However, the effect of myocardial perfusion abnormalities on regional diastolic dysfunction has not been established in patients with HCM. We investigated the relationship between regional myocardial perfusion abnormalities and regional early diastolic function in 31 patients with HCM and 8 control patients. Short-axis images of the left ventricle recorded by cine magnetic resonance imaging were divided into ten blocks. The time-to-peak-wall-thickness-thinning rate (TPWR) and the wall thickness were measured in each block. Of the 310 blocks from the patients with HCM, 242 (78%) showed normal thallium-201 uptake (group 1), 40 (13%) showed slightly decreased uptake (group 2), and 28 (9%) showed markedly decreased uptake (group 3). There was no difference in the regional wall thickness among the three groups. The TPWR was longer in patients with HCM than in control patients. It was significantly longer in group 3 (190+/-45ms) than in group 1 (167+/-36 ms) and group 2 (160+/-31 ms). (P < 0.01). The linear regression slope of the relationship between the TPWR and the regional wall thickness was significantly steeper in group 3 than in groups 1 and 2 (P < 0.05). In conclusion, abnormalities in regional myocardial perfusion, in addition to regional hypertrophy, contributed to the regional early diastolic dysfunction in patients with HCM.

Adult↗

Brain pharmacokinetics and in vivo receptor binding of 1,4-dihydropyridine calcium channel antagonists.

Brain pharmacokinetics of 1,4-dihydropyridine (DHP) calcium channel antagonists and their in vivo receptor binding in mice were characterized. The area under the concentration vs time curve (AUCbrain) for [3H]nifedipine, [3H]nimodipine and [3H]PN 200-110 in mouse brain after intravenous injection was higher than that for [3H]amlodipine. Brain/plasma concentration ratios (AUCbrain/AUCplasm) for [3H]nimodipine and [3H]PN 200-110 were 3 to 5 times higher than those for [ H]nifedipine and [3H]amlodipine. Further, brain/heart concentration ratios (AUCbrain/AUCheart) for [3H]nifedipine, [3H]nimodipine and [3H]PN 200-110 were about 20 times higher than the ratio for [3H]amlodipine. A significant amount of specific binding in particulate fractions of mouse brain was detected in vivo by intravenous injection of [3H]nifedipine, [3H]nimodipine and [3H]PN 200-110 but not [3H]amlodipine. These data suggest that [3H]nifedipine, [3H]nimodipine and [3H]PN 200-110 are more extensively taken up into brain from plasma than [3H]amlodipine and bind to the receptor sites in brain parenchymal cells in a significant amount in vivo. In conclusion, the present simultaneous measurement of pharmacokinetics and in vivo receptor binding in mouse brain suggests an usefulness of calcium channel antagonists such as nimodipine in the pharmacotherapy of brain diseases.

Animals↗

Responses of adrenal function to stimulation of lumbar and thoracic interspinous tissues in the rat.

In urethane-anesthetized Wistar rats, the responses of adrenal sympathetic nerve activity and catecholamine secretion were measured following chemical stimulation of lumbar and thoracic interspinous tissues. Injection of normal saline into the lower lumbar or lower thoracic interspinous tissues produced no changes in adrenal sympathetic nerve activity or catecholamine secretion. On the other hand, the injection of capsaicin produced protracted increases in adrenal nerve activity and catecholamine secretion both in CNS-intact animals and in animals acutely spinalized at the Cl-2 level. Repetitive electrical stimulation of the medial branch of a lumbar primary dorsal ramus, the nerve which provides sensation to the lumbar interspinous tissues, produced A- and C-reflex discharges, mediated at the spinal and supraspinal levels, in the adrenal sympathetic nerve.

Adrenal Glands↗

Concentration of global fallout 99Tc in rice paddy soils collected in Japan.

Analysis of global fallout 99Tc in environmental samples should provide useful information for predicting the nuclide behaviour under natural conditions which is important from the viewpoint of radioecology. Concentrations of 99Tc in rice paddy soils collected in Japan have been studied. After chemical separation, 99Tc in the final solution was measured by ICP-MS. The activity ratio of 99Tc/137Cs was used to understand the 99Tc behaviour in the environment because the fission yields of 99Tc and 137Cs from 235U or 239Pu are almost the same. The theoretical activity ratio from fission which is calculated now is about 3.0 x 10(-4). Our results showed that the range of activity ratios of 99Tc/137Cs in the soil samples was (2.0-5.2) x 10(-3); these ratios were one order of magnitude higher than the theoretical one. 99Tc has been accumulating in rice paddy soil like 137Cs has, although their mechanisms might differ. One of the reasons for the high ratio in the surface soil might be the ratios in the atmospheric samples, which have increased from the order of 10(-3) to 10(-2) (García-León et al., 1993).

Journal Article↗

Identification of HLA class II antigens as the targets of effector clones which may cause transfusion-associated graft-versus-host disease.

We established T cell clones, which were considered to be the possible cause of transfusion-associated graft-versus-host disease (TA-GVHD), from the peripheral blood lymphocytes (PBLs) of two patients. In both cases, several CD4+ cytotoxic T-cell (CTL) clones were established. In case I, the target antigen of the established CD4+ clones was a DRB1*0403-related antigen serologically typed as HLA DR4, which was one of the patient HLA antigens. In case II, the target of four out of five established CD4+ CTL was a DRB1*1302-related antigen. One CD4+ CTL clone showed cytotoxicity against cells carrying A*2402, B*4403, Cw*1403 and DPB1*0401. A monoclonal antibody (mAb) blocking study showed only anti-DP mAb inhibited the cytotoxicity of this clone. Thus, it might be considered that this clone recognizes HLA-DP with its binding peptides derived from either A*2402, B*4403, Cw*1403 or DRB1*1302. Our findings indicate that CD4+ CTLs may play important roles in the aetiology of TA-GVHD and that the antigens of patients recognized by donor-derived effector cells may not always recognize a single HLA antigen.

Aged↗

Long-term effect of manidipine on renal function and structure in uninephrectomized spontaneously hypertensive rats.

1. Long-term effects of manidipine hydrochloride (MAN), a calcium channel blocker, were examined in three groups of spontaneously hypertensive rats (SHR). Group 1 was given uninephrectomy (UNX) and MAN treatment, group 2 was given UNX and was not treated with MAN and group 3 was given neither UNX nor MAN treatment. 2. At week 15 after UNX, inulin clearance in group 1 rats decreased compared with rats in groups 2 and 3, but remained at the same level at week 40, when the level in group 2 rats declined below that in rats in groups 1 and 3. 3. Glomerular and tubulointerstitial lesions did not differ at week 15 after UNX among the three groups, whereas at week 40 both were advanced in the order of groups 2, 1 and 3. 4. Proteinuria did not differ between rats in groups 1 and 2 over the experimental period. 5. At week 15, the kidney weights of group 1 rats were greater than those of group 2 rats, indicating more prominent tubular hypertrophy in the former group. This was confirmed by morphometry of the proximal tubuli. In contrast, the glomerular volumes of rats in groups 1 and 2 were enlarged compared with that of rats in group 3, with no difference between the former two groups. 6. The findings suggest that MAN exerts renoprotective effects in SHR, both with regard to function and morphology. An effect on glomerular haemodynamics was considered to more likely be the mechanism underlying the renoprotective effect of MAN rather than that of a lowering of systemic blood pressure. 7. Augmented tubular hypertrophy after MAN treatment was an unexpected finding of the present study and the biological significance of this finding remains to be explored.

Animals↗

Spontaneous establishment of an Epstein-Barr virus-infected fibroblast line from the synovial tissue of a rheumatoid arthritis patient.

An Epstein-Barr virus (EBV)-infected fibroblast line, designated DSEK, was spontaneously established from synovial tissue of a patient with rheumatoid arthritis (RA). DSEK cells expressed EBV nuclear antigens EBNA-1 and EBNA-2 and latent membrane protein LMP-1. Cell surface markers of DSEK cells were similar to those of EBV-negative fibroblast clones derived from synoviocytes and were negative for lymphocyte and macrophage markers. DSEK cells expressed CD44, CD58, and HLA-DR antigens and spontaneously produced interleukin-10 basic fibroblast growth factor and transforming growth factor beta1. These results indicate that rheumatoid synoviocytes can be a target for EBV infection and suggest that EBV may play a role in the pathogenesis of RA.

Arthritis, Rheumatoid↗

Cloning of rat and mouse aquaporin-2 gene promoters and identification of a negative cis-regulatory element.

The promoters of rat and mouse aquaporin-2 (AQP-2) genes were cloned and compared with that of human genes. Nucleotide identity up to -593 bp was 62%, and consensus sequences such as TATA box and adenosine 3',5'-cyclic monophosphate responsive element were conserved. Deoxyribonuclease I footprint assay revealed a footprinted region at -210 to -184 bp in rat AQP-2 gene promoter produced by nuclear extract from nonexpressing (liver) tissue. The sequence of this region included a GATA motif but otherwise showed no homology with any other previously known cis-elements. Electromobility shift assay and ultraviolet cross-linking analysis confirmed that specific binding proteins to this element were present in kidney, spleen, and liver and that these proteins were distinct from GATA factors. Both deletion and mutation of this cis-element abolished the protein DNA binding and increased promoter activity in in vitro reporter gene assay using rat cultured hepatocyte Ac2F cells, suggesting the negative regulatory role of this cis-element. These results indicate that tissue-specific expression of AQP-2 gene may in part be regulated by this novel negative acting cis-element.

Animals↗

Multitracer studies on the accumulation of radionuclides in mushrooms.

We used the multitracer technique to study the transfer of several radionuclides to two mushroom species. Radionuclides accumulated in the fruiting bodies of the mushrooms in the order of 83Rb > 65Zn > 54Mn > 22Na > 75Se and 85Sr > 60Co > 88Y, 102Rh, 139Ce, 143.144Pm, 146.153Gd and 173Lu > 175Hf. The concentration ratio values for 83Rb, 65Zn and 54Mn in the fruiting bodies were more than 10, whereas those for 60Co, 88Y, 102Rh, 121mTe, 175Hf and the rare earth elements were less than 1. There were major differences in the accumulations of the alkali elements.

Basidiomycota↗

Regulated expression of endothelin converting enzymes in glomerular endothelial cells.

Endothelin converting enzyme (ECE) constitutes a potential regulatory site for the production of active mature endothelins. Two cDNAs (ECE-1 and -2) encoding ECE have recently been cloned, but the regulation of the expression of these ECE has not been clarified. In the study presented here, an attempt was made to determine whether or not glomerular endothelial cells (GEN) express ECE-1 and -2, and to learn how the expression of ECE-1 and -2 is regulated by kinase-mediated signaling systems. Ribonuclease protection assay revealed the expression of ECE-1 and -2 in cultured GEN, and the expression was increased approximately 2.5- and approximately 1.8-fold, respectively, by treatment with 10(-7) M 12-O-tetradecanocyl-phorbol-13-acetate (TPA) for 4 hours. These increases in ECE-1 and -2 expression with TPA were inhibited by cotreatment with calphostin C (10(-7) M). In contrast, 24-h treatment with 10(-7) M TPA significantly decreased the expression of ECE-1 and -2, indicating that the expression was tightly regulated by protein kinase C (PKC)-dependent mechanism(s). Actinomycin D (1 microgram/mL) abolished the TPA-induced increase of ECE-1 and -2 mRNA, whereas TPA treatment did not affect the mRNA stability of ECE-1 and -2, thus suggesting that TPA-induced increases of ECE-1 and -2 mRNA resulted from the transcriptional activation of ECE-1 and -2, gene, rather than from the increase of mRNA stability. In addition to the regulation by PKC, the effects of protein kinase A and G on ECE-1 and -2 expression were also examined. Treatment with chlorophenyl-thio cyclic AMP (200 microM) for 24 h decreased ECE-1 and -2 expression to approximately 50% and approximately 40% of the control value, respectively. 8-bromo-3', 5'-cyclic GMP also decreased ECE-1 and -2 expression to approximately 80% and approximately 25% of the control value, respectively. These results demonstrate that the expression of ECE-1 and -2 is regulated by kinase-mediated signaling systems, with the most prominent regulatory effect shown by protein kinase C.

Animals↗

Transcriptional regulation of aquaporin-2 water channel gene by cAMP.

Aquaporin-2 (AQP-2) water channel is a key molecule for urinary concentration whose expression is augmented by dehydration in vivo. To elucidate the regulatory mechanism of this phenomenon in vitro, mouse collecting duct cell lines were established from a transgenic mouse harboring temperature-sensitive simian virus 40 large T antigen gene and then screened for the AQP-2 expression, using ribonuclease protection assay. In one cell line designated C4, the endogenous AQP-2 mRNA level measured by ribonuclease protection assay increased fourfold after treatment with chlorophenylthio-cAMP (cpt-cAMP) (400 microM). In contrast, phorbol 12-myristate 13-acetate did not affect the AQP-2 mRNA level. To identify the molecular mechanism(s) of cAMP-induced upregulation of AQP-2 mRNA in C4 cells, luciferase assay was performed using various 5'-flanking regions of the human AQP-2 gene. Luciferase activity in C4 cells transfected with constructs containing approximately 2.8-kbp or 224-bp 5'-flanking region showed a 3.5-fold increase by cpt-cAMP treatment, indicating that the 224-bp 5'-flanking region contains the elements necessary for cAMP-induced regulatory mechanisms. This region contains cAMP-responsive element (CRE), and the deletion of the core sequence of CRE (GACGTCA) or introduction of mutation into CRE (GTGGTCA) completely abolished the responsiveness to cpt-cAMP, confirming the key role of CRE in the cAMP-induced transcriptional activation of the AQP-2 gene. Electrophoretic mobility shift assay revealed the existence of proteins binding to CRE in C4 cells and in rat kidney. The binding of CRE proteins to CRE was increased in the nuclear extract from cpt-cAMP-treated C4 cells and dehydrated rat kidney compared with those from controls. These results demonstrated that the CRE in the AQP-2 gene promoter is a key cis-element for cAMP-mediated transcriptional regulation of this gene and may be important for in vivo regulation of AQP-2 expression in a dehydrated state.

Animals↗

[Genetic diagnosis for post-transfusion graft-versus-host disease using PCR amplification of variable number of tandem repeats].

Post-transfusion graft-versus-host disease (PT-GVHD) is a potentially fatal adverse reaction to blood transfusion with a mortality rate of 95 percent or higher. We previously developed a definitive diagnostic test for PT-GVHD using polymerase chain reaction (PCR) amplification of highly polymorphic microsatellite repeat sequences. The results show the replacement of patient original DNA types with donor types on development of PT-GVHD. However, samples of patient blood obtained before transfusion are rarely stored until more than 2 weeks after the transfusion when PT-GVHD is usually suspected. On these occasions, we analyzed PCR amplified fragments of DNA extracted from fingernail clippings to determine the patient original DNA types. As a rapid and easily method, this system could be a useful tool for the clinical diagnosis of PT-GVHD.

Graft vs Host Disease↗

Highly diastereoselective synthesis of 2'-deoxy[2'-2H]ribonucleoside derivatives by the use of tris(trimethylsilyl)[2H]silane.

The use of (Me3Si)3SiH or (Me3Si)3Si2H-Et3B system in place of Bu3SnH or Bu3Sn2H-Et3B system for a reductive protylation or deuteration reaction of 2'-Br-2'-deoxy-3',5'-O-TIPDS-[2'-2H]ribonucleosides or 2'-Br-2'-deoxy-3',5'-O-TIPDS-ribonucleosides was confirmed to improve the reactions to afford an excellent diastereoselectivity, i.e., the diastereoselectivity of (2'S)- or (2'R)-2'-deoxy-[2'-2H]ribonucleosides obtained by the reaction at 0 degree C is equivalent to the diastereoselectivity obtained by that using Bu3SnH or Bu3Sn2H-Et3B system at < -70 degrees C; the reductive deuteration of 2'-O-phenoxythiocarbonyl-3',5'-O-TIPDS-ribonucleosides through (Me3Si)3Si2H-Et3B system at 0 degree C also gave (2'R)-2'-deoxy[2'-2H]ribonucleosides with such a high diastereo-selectivity as above.

Deoxyribonucleosides↗