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Biomedical subjects

Y Oda

Publications and source records attributed to Y Oda.

At least 253 records · Page 14Linked to original sources
PubMed1998

Binding characteristics of antibodies to the TSH receptor.

We have used fragments of the TSH receptor (TSHR) expressed in E. coli as glutathione S-transferase fusion proteins to produce rabbit polyclonal antibodies and a panel (n=5) of monoclonal antibodies to the extracellular fragment of the TSHR. The binding characteristics of the antibodies to linear, conformational, glycosylated and unglycosylated forms of the receptor in different assay systems have been investigated. The reactivity of these antibodies with the TSHR was assessed by Western blotting with both native and recombinant human TSHR expressed in CHO cells, immunoprecipitation of 35S-labelled full-length TSHR produced in an in vitro transcription/ translation system, immunoprecipitation of 125I-TSH/TSHR complexes, inhibition of 125I-TSH binding to the TSHR and fluorescence activated cell sorter (FACS) analysis of binding to CHO-K1 cells expressing the TSHR on their cell surface. Fab fragments of monoclonal antibodies were isolated, labelled with 125I and used to determine the affinity constants of these antibodies with receptor, bound and free Fab being separated by polyethylene glycol (PEG) precipitation. Rabbit polyclonal and mouse monoclonal antibodies reacted with the TSHR in Western blotting and one monoclonal antibody (3C7) was able to inhibit 125I-TSH binding to native human TSHR (74% inhibition), recombinant human TSHR (84% inhibition) and porcine TSHR (65% inhibition). Affinity constant values for TSHR monoclonal antibody Fab fragments calculated using Scatchard analysis were about 10(7) M(-1). Four out of five monoclonal antibodies reacted in FACS analysis with TSHR expressed on the surface of CHO-K1 cells. The FACS unreactive monoclonal (3C7) bound well to detergent solubilised TSH receptors and this emphasised the importance of using a combination of FACS analysis and radioactively-labelled probes in analysis of the TSH receptor. The monoclonal antibodies produced in this study were found to be of relatively low affinity but proved useful for detection of the receptor by Western blotting and by FACS analysis.

Animals↗
PubMed1998

Spinal meningeal melanocytoma presenting with superficial siderosis of the central nervous system. Case report and review of the literature.

Meningeal melanocytoma is a benign melanocytic tumor that originates most frequently from the melanocytes in the posterior fossa or along the spinal cord. This tumor generally occurs as an extraaxial mass that compresses adjacent neural structures to produce various neurological signs. The authors describe an unusual case in which a patient with a meningeal melanocytoma located at the thoracic spinal cord presented with superficial siderosis of the central nervous system (CNS). Extensive neuroradiological studies identified the presence of a spinal cord tumor, and postsurgical histological examination revealed the meningeal melanocytoma as a bleeding source. After surgery, lumbar puncture demonstrated normalization of the patient's cerebrospinal fluid; however, no neurological improvement occurred. The neurological deficits seem irreversible. Meningeal melanocytoma is biologically benign and can be cured by complete surgical resection; therefore, this tumor should be included in the differential diagnosis of pigmented lesions of the CNS. The authors reviewed 14 cases of well-documented meningeal melanocytoma in the literature and discuss the clinical, radiological, and pathological features of the present case to emphasize the importance of early diagnosis and identification of the source of bleeding in patients with superficial siderosis.

Brain Diseases↗
PubMed1998

[Validation of dissolution testing: evaluation of vibration levels of dissolution apparatuses].

The collaborative study participated by seven laboratories was carried out to develop a dissolution standard for evaluating vibration levels of dissolution apparatuses using enteric-coated granules of cefalexin (EG). Dissolution apparatuses could be divided into two groups according to their vibration levels and the dissolution test results of EG by the rotating basket method at 50 rpm. The critical value of acceleration was about 0.05 m/s2. The upper limit of normal dissolution rates of EG was calculated from the results of the rotating basket method at 50 rpm obtained from low vibration apparatuses. All high vibration apparatuses used in this study were distinguished by the limit from low vibration apparatuses, although most of them were not distinguished by current USP calibrators. These results suggest that EG would be useful as a calibrator for detection of apparatuses on high vibration levels.

Calibration↗
PubMed1998

Nuclear expression of YB-1 protein correlates with P-glycoprotein expression in human osteosarcoma.

The Y-box-binding protein, YB-1, is a member of the DNA-binding protein family. It binds to the Y-box, an inverted CCAAT box, in the promoter region of the human multidrug resistance 1 gene, which encodes P-glycoprotein (P-gp). Nuclear localization of YB-1 protein has been reported to be associated with the intrinsic expression of P-gp in human breast cancer. We studied the immunohistochemical expression of YB-1 protein in 69 untreated biopsy specimens of conventional osteosarcomas and compared it with the expression of P-gp. Furthermore, cell proliferation, as determined by the MIB-1-labeling index (MIB-1-LI), was measured by immunohistochemistry. In all 69 untreated osteosarcomas, YB-1 protein was expressed in the cytoplasm. In 32 of 69 (46%) cases, YB-1 was also localized in the nucleus. The expression of P-gp was evident in 23 of these 32 cases, and there was a significant correlation between the nuclear expression of YB-1 and P-gp expression (P < 0.0001). Chondroblastic osteosarcoma expressed YB-1 in the nucleus more frequently (eight of nine cases) than did other types of osteosarcoma, whereas P-gp was also frequently expressed in chondroblastic subtype. There was no correlation between the nuclear expression of YB-1 and histological grade. The MIB-1-LI was significantly higher in cases showing the nuclear expression of YB-1 (MIB-1-LI averaged 22.56 in cases with only cytoplasmic expression of YB-1 but averaged 28.20 in cases with cytoplasmic and nuclear expression of YB-1; P = 0.0477). In human osteosarcoma, nuclear localization of YB-1 protein was associated with the expression of P-gp, suggesting that YB-1 could be a prognostic marker for multidrug resistance in osteosarcoma.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1998

[Coronary artery fistula with left atrial myxoma: report of a case].

A 64-year-old male was referred for surgical treatment of left atrial myxoma. Preoperative coronary angiography revealed coronary artery fistula from the left anterior descending artery and the circumflex artery draining into the main pulmonary artery. Operative treatment was performed including resection of the myxoma, patch closure of the atrial septal defect, and closure of the fistula with pledgeted mattress sutures from within the main pulmonary artery on cardiopulmonary bypass. His postoperative course was uneventful, and disappearance of the left atrial myxoma and the coronary artery fistula was ascertained by echocardiography and coronary angiography.

Arterio-Arterial Fistula↗
PubMed1998

[Hemophagocytic syndrome associated with tuberculosis and mycoplasma infection in two patients].

Patient 1: A 48-year-old man was admitted to Osaka Red Cross Hospital because of fever and dyspnea. Laboratory examination revealed pancytopenia, liver dysfunction and hematostatic abnormality. Chest radiographs obtained on admission revealed ground-glass opacity in both lung fields, and an analysis of arterial blood showed severe hypoxemia (PaO2:46.8 Torr). Pulse therapy with methylprednisolone was started. Although the hypoxemia subsided and radiographic findings rapidly improved, pancytopenia persisted. Examination of bone marrow aspirate revealed mature histiocytes with marked hemophagocytosis. Amplified Mycobacterium tuberculosis direct tests of bronchoalveolar lavage fluid, sputum, urine, and bone marrow were all positive, and Mycobacterium tuberculosis was cultured from sputum and urine. Although the patient was taking antituberculous agents, his pancytopenia persosted. Treatment with etoposide induced remssion. Patient 2: A 19-year-old woman was admitted to Osaka Red Cross Hospital because of prolonged cough and fever. Laboratory examination revealed leukocytosis, liver dysfunction, and hematostatic abnormality. Serologic tests provided conclusive evidence of Mycoplasma infection and a CRP test was strongly positive. Chest radiographs obtained on admission revealed infiltration shadows in the middle and lower lung fields on both sides, with left pleural effusion. An analysis of arterial blood showed hypoxemia (PaO2: 54.2 Torr). Examination of bone marrow and pleural effusion samples revealed mature histiocytes with marked hemophagocytosis. Although treatment with antibiotics and pulse therapy with methylprednisolone was started, the patients respiratory functions deteriorated. Endotracheal intubation was performed. Therapy with etoposide induced remission. Hemophagocytic syndrome associated with Mycoplasma infection and tuberculosis appears to be exceedingly rare. In these 2 cases, it was difficult to achieve remission with therapy for the underlying infections, but etoposide treatment was effective.

Adolescent↗
PubMed1998

The effects of retraction forces applied to the anterior segment of orthodontic arch wires: differences in wire deflection with wire size.

Orthodontic arch wire deflection was studied in relationship to retraction forces applied to arch wires during anterior retraction with sliding mechanics. Two different sizes of orthodontic stainless steel arch wires (SS wires) .016" x .022" and .019" x .025", were examined. Retraction forces were generated using an elastic polymer material to measure arch wire deflection and applied to the arch wires by a commonly-used anterior retraction device with power chains hooked from the first molar areas to the pre-welded posts at distal site of the lateral incisors. The force application caused both the .016" x .022" and the .019" x .025" arch wires to deflect. The amount of deflection increased almost in proportion to the retraction force applied. With the same retraction force applied, the amount of deflection of the .019" x .025" wire was approximately 47.1% of that of the .016" x .022" wire.

Dental Arch↗
PubMed1997

Fluorometric assay of binding specificity of plant lectins to yeast cells by biotin-avidin system and its application to the classification of yeast cells.

A fluorometric assay of lectin binding to yeast cells is reported. The relative amount of biotinylated lectins bound to the yeast cells was estimated by enzyme activity using 4-methylumbelliferyl-beta-D-galactoside as a substrate for the lectin-bound beta-galactosidase through biotin-avidin interaction. Binding properties of 4 mannose-specific and 3 glucose/mannose-specific lectins to 22 different species of yeast cells were studied. The binding reaction of biotinylated lectins to the yeast cells was rapid and became constant within 10 min. Each lectin showed its characteristic binding specificity to each yeast species. The relative fluorescent intensities observed for 4-methylumbelliferone released by the action of bound beta-galactosidase were good indicators for the classification of yeast cells in quantitative base. We found that the yeast cells of the Saccharomyces genus can be classified into three groups, and those of Pichia were grouped into two groups. The present method can examine many samples simultaneously and be completed within 3 h.

Agglutination↗
PubMed1997

Tertiary and quaternary structures of 0.19 alpha-amylase inhibitor from wheat kernel determined by X-ray analysis at 2.06 A resolution.

The crystal structure of 0.19 alpha-amylase inhibitor (0.19 AI) from wheat kernel was determined by the multiple-isomorphous replacement method coupled with density modification and noncrystallographic symmetry averaging and then refined by simulated annealing using diffraction data to 2.06 A resolution (R = 18.7%, free R = 22.3%). The asymmetric unit has four molecules of 0.19 AI, each comprised of 124 amino acid residues. Electron density for residues 1-4 and 69-77 is absent in all subunits, probably because of the intrinsic flexibility of these segments. Each subunit has four major alpha-helices and one one-turn helix which are arranged in the up-and-down manner, maintaining the favorable packing modes of the alpha-helices. 0.19 AI, however, has two short antiparallel beta-strands. All 10 cysteine residues in 0.19 AI form disulfide bonds (C6-C52, C20-C41, C28-C83, C42-C99, and C54-C115), consistent with the assignments made biochemically for 0.28 AI from wheat kernel and by NMR analysis of the bifunctional alpha-amylase/trypsin inhibitor from ragi seeds (RBI). The disulfide bond patterns in these AIs are similar to those in the hydrophobic protein from soybean (HPS), which lack only the bond corresponding to C28-C83 in 0.19 AI. Extensive interactions occurred between particular pairs of 0.19 AI subunits, mainly involving hydrophobic residues. Comparisons of the structures of 0.19 AI, RBI, and HPS showed that the arrangements of the major alpha-helices are similar but the conformations of the remaining residues differ markedly. The present X-ray analysis for 0.19 AI and the NMR analysis for RBI suggest that all the AIs in this family have a common fold. The alpha-amylase binding site is discussed on the basis of the tertiary and quaternary structures of 0.19 AI together with biochemical and spectroscopic data for AIs.

Amino Acid Sequence↗
PubMed1997

Use of a binary mixture of quaternary ammonium salts in fluorometric determination of glycosaminoglycans.

The concentration of glycosaminoglycans, in an aqueous solution was determined by observing the fluorescent intensity of the ion of 2-hexadecyl-9H-pyrido[4,3b]indole at 450 nm by irradiating 253-nm light after dissolution of the insoluble salt in ethanol, which was formed by the reaction between a solution of a sample of glycosaminoglycan and an aqueous solution of the binary mixture of fluorescent 2-hexadecyl-9H-pyrido[4,3b]indolium bromide and hexadecyl pyridinium chloride. The fluorescent quaternary ammonium salt, which was slightly soluble in water, was solubilized in water by forming mixed micelles with a nonfluorescent soluble quaternary ammonium salt. The present method showed good linearity for the calibration curves between 5 and 1000 micrograms/ml in all samples of glycosaminoglycan except keratan sulfate. The relative standard deviation in determination was less than 3% for the whole calibration range.

Chondroitin Sulfates↗
PubMed1997

Efficient gene transduction by Epstein-Barr-virus-based vectors coupled with cationic liposome and HVJ-liposome.

We show here a novel non-viral strategy to transduce human cells by using an EBV-based vector system. The EBV-based vectors, the plasmid vectors carrying EBV oriP (origin for plasmid replication) and EBNA (EBV nuclear antigen) 1 gene from EBV genome, were combined with 2 gene delivery systems, i.e., cationic liposome and HVJ-liposome. By both methods, EBV-based vectors could be more efficiently transfected into HeLa cells than non-EBV, conventional plasmid vectors. When human primary fibroblasts were transfected, EBV-based vectors coupled with cationic liposome but HVJ-liposome resulted in successful gene transduction, while human bone marrow cells were transduced with both HVJ-liposome- and cationic liposome-EBV vectors. These results suggest the potential applications of the EBV-based vector system for gene therapy.

Bone Marrow Cells↗
PubMed1997

Alternatively spliced forms of the cGMP-gated channel in human keratinocytes.

Alternatively spliced forms of the alpha subunit of the cGMP-gated channel have been cloned from human keratinocytes. One form encodes a complete channel which is almost identical to the rod photoreceptor. A second spliced variant would encode a protein missing a portion of the intracellular hydrophilic domain and the putative first transmembrane domain. Both complete and spliced variants of the channel also were found in epidermis. The expression of the complete form of the channel was induced by levels of extracellular calcium which promote keratinocyte differentiation. The cGMP-gated channel may play an important role in calcium induced keratinocyte differentiation by mediating Ca2+ entry.

Alternative Splicing↗
PubMed1997

Binding mode of benzhydroxamic acid to Arthromyces ramosus peroxidase shown by X-ray crystallographic analysis of the complex at 1.6 A resolution.

The crystal structure of Arthromyces ramosus peroxidase (ARP) in complex with benzhydroxamic acid (BHA) as determined by X-ray analysis at 1.6 A shows unambiguously how BHA binds to ARP. BHA is located in the distal heme pocket. Its functional groups are held by three hydrogen bonds to His56N(epsilon), Arg52N(epsilon), and Pro(154)O, but are too far away to interact with the heme iron. The aromatic ring of BHA is positioned at the entrance of the channel to the heme pocket, approximately parallel to the heme group. Most water molecules at the active site of the native enzyme are replaced by BHA, leaving a ligand, probably a water molecule, at the sixth position of the heme. Results are compared with spectroscopic data.

Binding Sites↗
PubMed1997

Polycaprolactone depolymerase produced by the bacterium Alcaligenes faecalis.

Several microorganisms were isolated as bacteria degrading polycaprolactone (PCL), and one of them, a strain B273 identified as Alcaligenes faecalis, was selected. Because this strain produced only slight PCL depolymerase activity, the hyperproducing mutant, TS22, was isolated after UV irradiation. Synthesis of PCL depolymerase was derepressed, probably based on the altered regulation of metabolic pathways in strain TS22. The partially purified enzyme hydrolyzed p-nitrophenyl fatty acids and triglycerides other than PCL, but not poly(3-hydroxybutyrate), indicating that PCL depolymerase may be a kind of lipase.

Alcaligenes↗
PubMed1997

High-performance capillary electrophoresis of hyaluronic acid: determination of its amount and molecular mass.

The amount and the molecular mass of hyaluronic acid (HA) were determined by high-performance capillary electrophoresis. HA was observed at around 15 min by using an untreated fused-silica capillary (75 microns I.D.) of 58 cm length (effective length, 50 cm) at 20 kV in 50 mM phosphate buffer (pH 4.0). Calibration curves showed good linearity from 0.01 mg/ml to 3.3 mg/ml for all HA samples examined. The lower limit of detection by monitoring the absorbance at 185 nm was 1.0 microgram/ml at the signal-to-noise ratio of 5. HA samples were examined in a buffer containing pullulan (PU) as an additive for the matrix formation material. The HA samples showed marked peak-broadening when analyzed in the buffer solution containing PU with a specified molecular mass. The peak broadening was based on the dispersion of the molecular mass of the HA sample analyzed.

Buffers↗
PubMed1997

Simultaneous quantitative determination method for sphingolipid metabolites by liquid chromatography/ionspray ionization tandem mass spectrometry.

Sphingolipid metabolites ceramide, sphingomyelin, sphingosine, psycosine, sphingosylphosphorylcholine, and dimethylsphingosine were separated and simulataneously quantitated by liquid chromatography/ionspray ionization tandem mass spectrometry (LC/MS/ MS). The use of glassware throughout minimized losses due to adsorption and the pretreatment of this method consisted of simple liquid-liquid extraction procedure with a mixture of chloroform and methanol. After separation on a short C18 silica column eluted in a gradient mode, the metabolites were detected by MS/ MS. This assay allows simultaneously quantification of these metabolites over a range of at least 0.1 to 100 ng/ 10(6) cells. The LC/MS/MS analyses took 10 to 15 min per sample and we could examine up to 50 samples per day. We also detected endogenous sphingosine 1-phosphate in HL-60 cells. The utility of the method was demonstrated by examining changes in metabolites levels in HL-60 cells after treatment with sphingomyelinase. It was found that sphingomyelinase from Bacillus cereus may have selectivity for acyl chain length.

Ceramides↗
PubMed1997

A phylogenetic analysis of Saccharomyces species by the sequence of 18S-28S rRNA spacer regions.

Sequences of two internally transcribed spacer regions between 18S and 28S rRNA genes were determined to assess the phylogenetic relationship in the strains belonging to the genus Saccharomyces. The sequences of S. bayanus and S. pastorianus were quite similar, but not identical. Two phylogenetic trees constructed by the neighbor-joining method showed that all the species examined were distinguished from one another. The Saccharomyces sensu stricto species: S. cerevisiae, S. bayanus, S. paradoxus and S. pastorianus, were closely related and far from the Saccharomyces sensu lato species including S. barnetti, S. castellii, S. dairensis, S. exiguus, S. servazzii, S. spencerorum and S. unisporus, and an outlying species, S. kluyveri.

DNA, Fungal↗