Biomedical subjects

Y Kubo

Publications and source records attributed to Y Kubo.

At least 55 records · Page 3Linked to original sources

PubMed2001

Os odontoideum associated with hypertrophic ossiculum terminale. Case report.

The authors report on the case of a 20-year-old man who presented with a transient tetraparesis. Neuroimaging studies demonstrated atlantoaxial dislocation and ventral compression of the rostral spinal cord caused by a quite rare association of os odontoideum and hypertrophic ossiculum terminale. The patient underwent removal of two free ossicula via a transoral approach and posterior fusion in which an autogenous bone graft was placed. The majority of cases of os odontoideum are believed to be an acquired form; however, controversy with regard to the congenital causes of os odontoideum remains. One hypothesis is that os odontoideum results from the failure of fusion and the hypertrophy of the proatlas, although considerable confusion surrounds this hypothesis because definitive classification of os odontoideum-to differentiate between similar anomalies-has not been established. This rare coincidence in the current case supports the belief that os odontoideum has a different embryological origin from ossiculum terminale, which is thought to be a proatlantal remnant.

Abnormalities, Multiple↗

PubMed2001

Non-invasive approach for diagnosing atrophic gastritis using the 13C-bicarbonate breath test.

The experiments presented here were done to evaluate whether the levels of CO2 in respiratory air during the 13C-bicarbonate breath test (13C-BBT) may be used as a marker of non-invasive diagnosis of the levels of atrophic gastritis. Twenty-eight patients with chronic gastritis and five healthy volunteers were enrolled in the study. Moreover, experimental gastritis was induced in rats by N-methy-N-nitro-N-nitrosoguanidine. In human, the levels of atrophic gastritis were evaluated from the vascular pattern of the gastric fornix. Total delta 13CO2 calculated from the 13C-BBT and the mucosal thickness ratio (MTR) were measured in rats with experimental gastritis. The levels of 13CO2 were significantly higher from patients with a vascular pattern at the fornix than in those without a vascular pattern (p<0.01). There was a good correlation between MTR and the levels of 13CO2, in rats with experimental gastritis (p<0.01). These findings indicate that the levels of 13CO2 during 13C-BBT reflect the levels of atrophic gastritis and show its clinical significance for non-invasive evaluation of atrophic gastritis. This has important clinical implications in selecting Helicobacter pylori-positive cases for therapy and follow-up.

Age Factors↗

PubMed2001

Economic analysis of a filmless system based on the hospital information system.

A filmless system (FLS) based on a picture archiving and communication system (PACS) equipped with UNIX work- stations and a local area network (LAN) specialized for viewing radiographs has not been deemed acceptable, for reasons of economics. However, personal computers (PCs) have recently become more powerful, to the point where PCs approach UNIX workstations in terms of capabilities. As a result, a PC-based image viewing workstation (IVW) has sufficient functions for practical use. Diagnostic resolution is not equal to that of film, but is comparable. Since the hospital information system (HIS) includes many PC terminals connected by a LAN, the cost problem can be resolved by using these PC terminals as the IVWs. In order to investigate the practicability of this idea, two types of FLSs using HIS facilities were designed: one is a system based on the use of high-resolution cathode ray tubes (H-CRTs), and the other is based on the use of conventional CRTs and radiologist reports, minimizing the number of H-CRTs. The total costs of the two systems were analyzed. As a result, the former FLS was found to be about 15% more expensive than the latter, which was less expensive than a film-based system (FBS). However, whether the FLS is more profitable than the FBS from the viewpoint of hospital management strongly depends on the medical insurance system.

Costs and Cost Analysis↗

PubMed2001

Primary structure, developmental expression and functional properties of an inward rectifier K+ channel of the tunicate.

A cDNA encoding for a tunicate inward rectifier K+ channel (TuIRK) was isolated. TuIRK exhibited the highest similarity (approximately 50%) with mammalian Kir2 (IRK) subfamily. Maternal RNA of TuIRK was detected by RT-PCR in unfertilized eggs. By in situ hybridization, the transcript was observed at the 32-cell stage, restricted at the 64-cell stage in anterior epidermal cells of a4-2 blastomere lineage, and disappeared at the late gastrula stage. Therefore, TuIRK was identified to be the inward rectifier whose expression was previously reported to change dramatically upon the neural/epidermal cell fate selection. In Xenopus oocytes, TuIRK expressed a strongly inward rectifying K+ current. The basic electrophysiological properties of TuIRK were similar to those of the mouse IRK1 (mIRK1), except that the sensitivity to the block by extracellular Mg2+ was much lower than that of mIRK1. To identify the structural determinant, we made mutants of the pore region, and then of the extracellular loop (N226 of TuIRK, and E125 of mIRK1). In E125N mutant of mIRK1, the sensitivity to the Mg2+ block was decreased significantly, whereas N226E of TuIRK1 did not acquire the sensitivity. These results demonstrate the contribution of this site to the Mg2+ block and the presence of additional determinant(s).

Amino Acid Sequence↗

PubMed2001

[Effect of extracellular Ca2+ on neuronal function].

Metabotropic glutamate receptors (mGluR) are well known to have a significance for the plastic change of synaptic neurotransmission. We observed that mGluR can be activated not only by glutamate but also by extracellular Ca(2+) of physiological concentration. Our observation suggests that extracellular Ca(2+) plays a critical role for the neuronal function, not only by serving as a source to elevate intracellular Ca(2+) concentration by entering through various channels, but also by activating metabotropic glutamate receptors as a ligand.

English Abstract↗

PubMed2000

Regulator of G protein signaling 8 (RGS8) requires its NH2 terminus for subcellular localization and acute desensitization of G protein-gated K+ channels.

Functional roles of the NH(2)-terminal region of RGS (regulators of G protein signaling) 8 in G protein signaling were studied. The deletion of the NH(2)-terminal region of RGS8 (DeltaNRGS8) resulted in a partial loss of the inhibitory function in pheromone response of yeasts, although Galpha binding was not affected. To examine roles in subcellular distribution, we coexpressed two fusion proteins of RGS8-RFP and DeltaNRGS8-GFP in DDT1MF2 cells. RGS8-RFP was highly concentrated in nuclei of unstimulated cells. Coexpression of constitutively active Galpha(o) resulted in translocation of RGS8 protein to the plasma membrane. In contrast, DeltaNRGS8-GFP was distributed diffusely through the cytoplasm in the presence or absence of active Galpha(o). When coexpressed with G protein-gated inwardly rectifying K(+) channels, DeltaNRGS8 accelerated both turning on and off similar to RGS8. Acute desensitization of G protein-gated inwardly rectifying K(+) current observed in the presence of RGS8, however, was not induced by DeltaNRGS8. Thus, we, for the first time, showed that the NH(2) terminus of RGS8 contributes to the subcellular localization and to the desensitization of the G protein-coupled response.

Active Transport, Cell Nucleus↗

PubMed2000

NK-mediated elimination of mutant lymphocytes that have lost expression of MHC class I molecules.

Mutant cells generated in vivo can be eliminated when mutated gene products are presented as altered MHC/peptide complexes and recognized by T cells. Diminished expression of MHC/peptide complexes enables mutant cells to escape recognition by T cells. In the present study, we tested the hypothesis that mutant lymphocytes lacking expression of MHC class I molecules are eliminated by autologous NK cells. In H-2b/k F1 mice, the frequency of H-2Kb-negative T cells was higher than that of H-2Kk-negative T cells. The frequency of H-2K-deficient T cells increased transiently after total body irradiation. During recovery from irradiation, H-2Kk-negative T cells disappeared more rapidly than H-2Kb-negative T cells. The disappearance of H-2K-deficient T cells was inhibited by administration of Ab against asialo-GM1. H-2Kk-negative T cells showed higher sensitivity to autologous NK cells in vitro than H-2Kb/k heterozygous or H-2Kb-negative T cells. Adding syngeneic NK cells to in vitro cultures prevented emergence of mutant cells lacking H-2Kk expression but had little effect on the emergence of mutant cells lacking H-2Kb expression. Results in the H-2b/k F1 strain correspond with the sensitivity of parental H-2-homozygous cells in models of marrow graft rejection. In H-2b/d F1 mice, there was no significant difference between the frequencies of H-2Kb-negative and H-2Kd-negative T cells, although the frequencies of mutant cells were different after radiation exposure among the strains examined. H-2b/d F1 mice also showed rapid disappearance of the mutant T cells after irradiation, and administration of Ab against asialo-GM1 inhibited the disappearance of H-2K-deficient T cells in H-2b/d F1 mice. Our results provide direct evidence that autologous NK cells eliminate mutant cell populations that have lost expression of self-MHC class I molecules.

Animals↗

PubMed2000

Enzymatic synthesis of 1,3,6,8-tetrahydroxynaphthalene solely from malonyl coenzyme A by a fungal iterative type I polyketide synthase PKS1.

The Colletotrichum lagenarium PKS1 gene encoding iterative type I polyketide synthase of 1,3,6,8-tetrahydroxynaphthalene (T4HN) was overexpressed in Aspergillus oryzae. SDS-PAGE analysis of the cell-free extract prepared from the transformant showed an intense band of 230000 which corresponded to the molecular weight of the deduced PKS1 protein. By using this cell-free extract, in vitro synthesis of T4HN was successfully confirmed as the first example of the fungal multi-aromatic ring polyketide synthase activity ever detected. To identify the starter unit for T4HN synthesis, (14)C-labeled acetyl CoA and/or (14)C-labeled malonyl CoA were used as substrates for T4HN synthase reaction. Observed was the incorporation of (14)C label into T4HN solely from malonyl CoA even in the absence of acetyl CoA and not from acetyl CoA. This in vitro result unambiguously identified that malonyl CoA serves as the starter as well as extender units in the formation of T4HN by fungal polyketide synthase PKS1.

Aspergillus oryzae↗

PubMed2000

Superconducting fluctuations and the pseudogap in the slightly overdoped high- T(c) superconductor TlSr2CaCu2O6.8: high magnetic field NMR studies

From measurements of the 63Cu Knight shift ( K) and the nuclear spin-lattice relaxation rate ( 1/T1) under magnetic fields from zero up to 28 T in the slightly overdoped high- T(c) superconductor TlSr2CaCu2O6.8 ( T(c) = 68 K), we find that the pseudogap behavior, i.e., the reductions of 1/T1T and K above T(c) from the values expected from the normal state at high T, is strongly field dependent and follows a scaling relation. We show that this scaling is consistent with the effects of the Cooper pair density fluctuations. The present finding contrasts sharply with the pseudogap property reported previously in the underdoped regime where no field effect was seen up to 23.2 T. The implications are discussed.

Journal Article↗

PubMed2000

Cysteine-scanning mutagenesis around transmembrane segments 1 and 11 and their flanking loop regions of Tn10-encoded metal-Tetracycline/H+ antiporter.

Putative transmembrane helices (TM) 1 and 11 in the metal-tetracycline/H(+) antiporter are predicted to be close to each other on the basis of disulfide cross-linking experiments of the double-cysteine mutants in the periplasmic loop regions (Kubo, Y., Konishi, S., Kawabe, T., Nada, S., and Yamaguchi, A. (2000) J. Biol. Chem. 275, 5270-5274). In this study, each amino acid from Asn-2 to Gly-44 in the putative TM1 and loop1-2 regions or that from Ser-328 to Gly-366 in TM11 and its flanking regions was individually replaced with cysteine. With respect to the TM1 region, 10 mutants, from T5C to L14C, were all not reactive with N-ethylmaleimide (NEM), and from D15C to I22C, NEM-reactive and non-reactive mutations periodically appeared every two residues. Three mutants, M23C to V25C, were all NEM-reactive, but the degree of the latter two mutants was very low. Seven mutants, from L26C to E32C, were all highly reactive with NEM. Therefore, the region of TM1 is composed of the 21 amino acid residues from Thr-5 to Val-25. It is a partially amphiphilic helix, that is, the N-terminal (cytoplasmic) half is embedded in the hydrophobic interior, and the C-terminal (periplasmic) half faces a water-filled channel. With respect to TM11, nine mutants, from S328C to G336C, and six mutants, from L361C to G366C, were all reactive with NEM. On the other hand, out of the 24 mutants, from L337C to S360C, 17 were not reactive with NEM, and the 7 NEM-reactive mutants were scattered, indicating that this region is a transmembrane segment. The 7 residues from Val-347 to Phe-353 including Pro-350 formed a central hydrophobic core, and the 7 NEM-reactive mutations were periodically distributed in its flanking regions, indicating that both ends of TM11 face a water-filled channel. Ala-354 is located at about 1/3 of the length from the periplasmic end of TM11. Disulfide cross-linking experiments on double-cysteine mutants having the combination of A354C and a cysteine-scanning mutation in the loop1-2 region indicated that loop1-2 is very flexible and close to the periplasmic end of TM11. Tetracycline prevented the cross-linking formation between the periplasmic ends of TM1 and TM11; however, it did not affect the cross-linking between loop1-2 and TM11, indicating that the substrate-induced conformational change involves a shift in the relative locations of TM1 and TM11.

Amino Acid Sequence↗

PubMed2000

Proximity of periplasmic loops in the metal-Tetracycline/H(+) antiporter of Escherichia coli observed on site-directed chemical cross-linking.

Our previous study on second-site suppressor mutations of the Tn10-encoded metal-tetracycline/H(+) antiporter suggested that Leu(30) and Ala(354), located in periplasmic loop 1-2 and 11-12, respectively, are conformationally linked to each other (Kawabe, T., and Yamaguchi, A. (1999) FEBS Lett. 457, 169-173). To determine the spatial proximity of these two residues, cross-linking gel-shift assays of the L30C/A354C double mutant were performed after the mutant had been oxidized with Cu(2+)/o-phenanthroline. The results indicated that Leu(30) and Ala(354) are close to each other but that Gly(62), which is located in cytoplasmic loop 2-3, and Ala(354) are distant from each other, as a negative control. Then, a single Cys residue was introduced into each of the six periplasmic loop regions (P1-P6), and eleven double mutants were constructed. Of these eleven double Cys mutants, the L30C/A354C and L30C/T235C mutants showed a mobility shift on oxidation, indicating that P1 is spatially close to P4 as well as P6. In contrast, the other nine mutants, L30C/S92C, L30C/S156C, L30C/S296C, S92C/S296C, S92C/T235C, S92C/A354C, S156C/T235C, S156C/S296C, and S156C/A354C, showed no mobility shift under oxidized conditions on intramolecular cross-linking. The S92C and S296C mutants showed dimerization on intermolecular cross-linking, indicating that P2 and P5 are located at the periphery of the helix bundle.

Amino Acid Sequence↗

PubMed2000

The transition state in the folding-unfolding reaction of four species of three-disulfide variant of hen lysozyme: the role of each disulfide bridge.

The effects of lacking a specific disulfide bridge on the transition state in folding were examined in order to explore the folding-unfolding mechanism of lysozyme. Four species of three-disulfide variant of hen lysozyme (3SS-lysozyme) were prepared by replacing two Cys residues with Ala or Ser: C6S/C127A, C30A/C115A, C64A/C80A and C76A/C94A. The recombinant hen lysozyme was studied as the standard reference containing four authentic disulfide bridges and the extra N-terminal Met: the recombinant hen lysozyme containing the extra N-terminal. Folding rates were measured by monitoring the change in fluorescence intensity associated with tri-N-acetyl-d-glucosamine binding to the active site of refolded lysozyme. It was confirmed that the folding rate of the recombinant hen lysozyme containing the extra N-terminal was the same as that of wild-type lysozyme, and that the folding rate was little affected by the presence of tri-N-acetyl-d-glucosamine (triNAG). The folding rate of C64A/C80A was found to be the fastest and almost the same as that of the recombinant hen lysozyme containing the extra N-terminal, and that of C30A/C115A the second, and that of C6S/C127A the third. The folding rate of C76A/C94A was particularly slow. On the other hand, the unfolding rates which were measured in the presence of triNAG showed the dependence on the concentration of triNAG. The intrinsic unfolding rate in the absence of triNAG was determined by extrapolation. Also in the unfolding rate, C76A/C94A was markedly slower than the others. It was found from the analysis of binding constants of triNAG to C64A/C80A during the unfolding process that the active site of C64A/C80A partly unfolds already prior to the unfolding transition. On the basis of these kinetic data, we suggest that C64A/C80A folding transition can occur with leaving the loop region around SS3 (C64-C80) flexible, while cross-linking by SS4 (C76-C94) is important for the promotion of folding, because it is an indispensable constraint on the way towards the folding transition state.

Amino Acid Substitution↗

PubMed2000

SiO(x) Monolayer Overcoating Effect on the TiO(2) Photocatalytic Oxidation of Cetylpyridinium Bromide.

The effect of SiO(x) monolayer coverage on the rate of TiO(2) photocatalytic oxidation of cetylpyridinium bromide (CPB) in aqueous solutions has been studied. The rate of CPB removal from the solution (5<pH<7) increases with the surface modification at concentrations below 4.5x10(-4) M and its promoting effect is enhanced with decreasing concentration. A modified Langmuir-Hinshelwood model, where the rate of surface reaction is assumed to be proportional to the coverage of CPB at the photostationary state is proposed for dilute reaction systems. As a result of the kinetic analyses, the acceleration of the reaction with the SiO(x) monolayer coverage can be attributed to the increase in the rate of adsorption due to the electrostatic attraction of cetylpyridinium ion. The suppression of Br(-) adsorption is also suggested as a minor contribution. Copyright 2000 Academic Press.

Journal Article↗

PubMed2000

Revascularization of canine cryopreserved tracheal allografts.

BACKGROUND: We examined the blood supply of a cryopreserved tracheal allograft and its morphohistologic changes after transplantation. METHODS: In each of 22 dogs, a five-ring tracheal segment was replaced by one of the following tracheal grafts: fresh autografts (n = 8), cryopreserved tracheal allografts (n = 8), or fresh allografts (n = 6). The cryopreserved tracheal allografts were preserved at -196 degrees C for 60 days. No immunosuppressant was given to any of the animals. All grafts were retrieved at 1 and 12 weeks and assessed by microangiography and histology. RESULTS: The epithelial denudation and the revascularization of the transverse intercartilaginous arteries were recognized within 7 days as common to each of the three types of grafts. In the cryopreserved tracheal allografts, neither cartilage degradation nor graft shrinkage occurred at 7 days. However, the recanalized transverse intercartilaginous arteries completely disappeared at 12 weeks, and marked shrinkage occurred; the cartilage cells were accompanied by karyolysis and were significantly decreased in number (p < 0.05). Recanalization of the transverse intercartilaginous arteries was also demonstrated in the fresh allografts; however, necrosis abruptly occurred as a result of acute rejection responses. CONCLUSIONS: Cryopreservation of a tracheal allograft provided sufficient reduction of the acute rejection responses, and blood supply to the cryopreserved tracheal allograft was established through the recanalized transverse intercartilaginous arteries within 7 days; however, subsequent chronic rejection responses resulted in occlusion of the transverse intercartilaginous arteries and atrophy.

Animals↗

PubMed2000

Anastomotic complications after bronchoplastic procedures for nonsmall cell lung cancer.

BACKGROUND: Anastomotic complications associated with bronchoplastic procedures cannot be completely avoided despite the improvements made in surgical techniques and suture materials. Thus, the present study attempted to clearly define the significant factors influencing anastomotic complications. METHODS: Between 1978 and 1998, 47 patients with primary nonsmall cell lung cancer underwent bronchoplastic procedures. The incidences of anastomotic complications were calculated according to each of the following clinical factors: primary site, age, pathologic type, pT factor, pN factor, pulmonary arterioplasty, surgical procedure, suture material, coverage of the anastomotic line, positive resection margin, and preoperative chemotherapy. The results were analyzed using univariate and multiple logistic regression analysis. RESULTS: Anastomotic complications occurred in 8 patients. Four had anastomotic dehiscence and 4 had stenosis. Of these 8 patients, the resection margin was diagnosed as being positive in 6 patients. Three showed metastasis of the most distal mediastinal lymph node whereas the others had a residual tumor at the bronchial resection margin. According to multiple logistic regression analysis, only pN factor (p = 0.04) and positive resection margin (p = 0.02) had a significant influence on the complications. CONCLUSIONS: Thus, pN2 patients, especially those with metastasis of the most distal mediastinal lymph node and patients with a residual tumor at the bronchial resection margin, have a significantly higher risk of anastomotic complications.

Adult↗

PubMed2000

Effect of vitamin A palmitate on vitamin A-deficient rabbits

Purpose: We examined the effects of vitamin A palmitate (VA pal) eyedrops on the symptoms caused by vitamin A-deficiency in rabbits. Methods: Three-week-old rabbits were raised on a vitamin A-deficient diet, and were examined for the quantity of retinol in the serum and the condition of the anterior segment of the eye. The vitamin A-deficient animals were treated with VA pal eyedrops.Results: The retinol in the serum began to decrease 7 months after the animals were placed on the vitamin A-deficient diet. After 10 months, superficial punctate keratitis and the loss of conjunctival goblet cells were observed. Treatment of the disease in the anterior segment of the eyes with VA pal eyedrops resulted in restoration of normal condition within 3 weeks. The treatment increased the goblet cells in the ocular conjunctiva and the retinol in the serum. Conclusions: These results show the efficacy of topical VA pal treatment for vitamin A-deficient rabbits.

Journal Article↗

PubMed2000

Flavonols from Heterotheca inuloides: tyrosinase inhibitory activity and structural criteria.

Tyrosinase inhibitory activity of flavonols, galangin, kaempferol and quercetin, was found to come from their ability to chelate copper in the enzyme. In contrast, the corresponding flavones, chrysin. apigenin and luteolin, did not chelate copper in the enzyme. The chelation mechanism seems to be specific to flavonols as long as the 3-hydroxyl group is free. Interestingly, flavonols affect the enzyme activity in different ways. For example, quercetin behaves as a cofactor and does not inhibit monophenolase activity. On the other hand, galangin inhibits monophenolase activity and does not act as a cofactor. Kaempferol neither acts as a cofactor nor inhibits monophenolase activity. However, these three flavonols are common to inhibit diphenolase activity by chelating copper in the enzyme.

Arnica↗

PubMed2000

Molecular design of antibrowning agents.

Tyrosinase inhibitory and antioxidant activity of gallic acid and its series of alkyl chain esters were investigated. All inhibited the oxidation of L-3,4-dihydroxyphenylalanine (L-DOPA) catalyzed by mushroom tyrosinase. However, gallic acid and its short alkyl chain esters were oxidized as substrates yielding the colored oxidation products. In contrast, the long alkyl chain esters inhibited the enzyme activity without being oxidized. This indicates that the carbon chain length is associated with their tyrosinase inhibitory activity, presumably by interacting with the hydrophobic protein pocket in the enzyme. On the other hand, the esters, regardless their carbon chain length, showed potent scavenging activity on the autoxidation of linoleic acid and 1,1-diphenyl-2-p-picryhydrazyl (DPPH) radical, suggesting that the alkyl chain length is not related to the activity. The effects of side-chain length of gallates in relation to their antibrowning activity are studied.

Agaricales↗