Extended-wear soft contact lenses don't change the ocular flora.
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Aged↗
Biomedical subjects
T Ohshima
Publications and source records attributed to T Ohshima.
Autotransfusion supported by erythropoietin therapy in transurethral resection of the prostate.
We investigated the collection and transfusion of autologous blood after treatment with EPO in 68 BPH patients (including 10 controls) who were scheduled to undergo TUR-P. All patients received oral and/or intravenous iron supplements. Assessments were made based on the preoperative increase in blood hemoglobin levels including autologous blood predonation (deltaHb). The deltaHb in patients undergoing collection of 600 ml of blood were as follows: control group: -0.36 +/- 0.57 g/dl; EPO group, 9 x 3000 units intravenously: 1.15 +/- 0.83 g/dl; EPO group, 6 x 6000 units intravenously: 0.79 +/- 0.80 g/dl; EPO group, 3 x 12,000 units subcutaneously: 1.47 +/- 0.62 g/dl. In patients undergoing collection of 800 ml of blood, the results were as follows: EPO group, 3 x 12,000 units subcutaneously: 1.80 +/- 0.69 g/dl; EPO group, 3 x 24,000 units subcutaneously: 2.03 +/- 0.77 g/dl. All EPO-treated patients successfully underwent surgery using their own blood, and none of them required homologous transfusion. The increase of Hb was greater in the patients treated with EPO than in controls, allowing safe preoperative blood collection even in elderly patients. In patients with relatively severe BPH, homologous transfusion could be avoided and surgery was performed safely.
Aged↗
Induction of apoptosis in vitro and in vivo by H-1 parvovirus infection.
Apoptosis induced by H-1 parvovirus infection was investigated in C6 rat glioblastoma cells and in newborn rats. Apoptotic changes, such as chromatin condensation, the appearance of apoptotic nuclear bodies and oligonucleosomal DNA ladders, were observed in infected C6 cells 2 days after infection. Inhibitor assay results suggest that a caspase-3-dependent apoptosis activation pathway is induced by H-1 virus infection in C6 cells. Observations made in vivo revealed that the number of apoptotic cells increased in the infected cerebellum, coinciding with known virus infection sites.
Animals↗
Effect of sequence of administration on the pharmacokinetic interaction between the anticholinergic drug biperiden and [3H]quinuclidinyl benzylate or [3H]N-methylscopolamine in rats.
In rats the pharmacokinetic interactions between the anticholinergic drug biperiden and [3H]quinuclidinyl benzylate ([3H]QNB) or [3H]N-methylscopolamine ([3H]NMS) is affected by the sequence in which the drugs are administered. Drug concentrations in various tissues were determined after intravenous administration of [3H]QNB or [3H]NMS (325 ng kg(-1)). Biperiden (6.4 mg kg(-1)) was administered either 5 min before, concomitantly with or 20 min after injection of [3H]QNB or [3H]NMS. When biperiden was administered concomitantly with or before [3H]QNB, distribution of [3H]QNB among the regions of the brain and other tissues was reduced; at 4 h the ratio of the distribution of [3H]QNB for experimental animals to that for control animals ranged from 0.15 to 0.9. When biperiden was administered after [3H]QNB, the distribution of [3H]QNB in the brain and other tissues was significantly higher than for the other two treatments (P < 0.01). However, for [3H]NMS the sequence of administration had no effect on the distribution of the drug in the brain and other tissues except for the kidney. In-vitro, in crude synaptosomal membranes, the amount of [3H]QNB at 2 h relative to the control concentration at equilibrium was 87% when biperiden was added before [3H]QNB and 56% when biperiden was added after [3H]QNB. In both instances the concentration of [3H]NMS reached equilibrium within 30 min. These findings suggest that the difference between the rate constant of association and dissociation at the possible site of action gives rise to the effect of the sequence of administration on the pharmacokinetic interaction.
Animals↗
[Evaluation of a synthetic peptide based species specific EIA kit for detection of antibodies to Chlamydia trachomatis with clinical specimens].
We have evaluated a new kit, PEPTIDE Chlamydia (Meiji Milk Products Co., Ltd, Tokyo), for detecting species specific antibodies to Chlamydia trachomatis with synthetic peptide as an antigen. Serum samples from women with C. trachomatis cervicitis (n = 45), healthy pregnant women (n = 100), and children suffering from C. pneumoniae lower respiratory tract infection (n = 9) were used. We have measured the serum IgG or IgA antibodies to C. trachomatis of those sera with PEPTIDE, Sero IPALIZA Chlamydia (Savyon Diagnostics, Israel), and HITAZYME Chlamydia (Hitachi Chemical Co., Ltd, Hitachi). Serum samples with discrepant results were further analyzed by a microimmunofluorescence test and immunoblotting (western blotting). IgG and IgA serum positive rate to C. trachomatis of sera from women with C. trachomatis cervicitis were similar in three kits, that is, 91% and 80% in PEPTIDE, 89% and 82% in Sero IPALIZA, and 84% and 76% in HITAZYME, respectively. IgG and IgA serum positive rate to C. trachomatis of sera from healthy pregnant women were 18% and 9% in PEPTIDE, 12% and 15% in Sero IPALIZA, and 15% and 13% in HITAZYME, respectively. Serum antibodies to C. trachomatis in serum samples from children with C. pneumoniae infection were all negative in PEPTIDE, but falsely positive in several cases in Sero IPALIZA or HITAZYME. In sera with discrepant results, PEPTIDE gave more identical results with a micro-IF test and immunoblotting analysis than Sero IPALIZA or HITAZYME. These results indicate that PEPTIDE is an useful kit to detect more species specific antibodies to Chlamydia trachomatis than former kits.
Antibodies, Bacterial↗
Enzymological characteristics of the hyperthermostable NAD-dependent glutamate dehydrogenase from the archaeon Pyrobaculum islandicum and effects of denaturants and organic solvents.
NAD-dependent glutamate dehydrogenase (L-glutamate:NAD oxidoreductase, deaminating; EC 1.4.1.2) was purified to homogeneity from a crude extract of the continental hyperthermophilic archaeon Pyrobaculum islandicum by two successive Red Sepharose CL-4B affinity chromatographies. The enzyme is the most thermostable NAD-dependent dehydrogenase found to date; the activity was not lost after incubation at 100 degrees C for 2 h. The enzyme activity increased linearly with temperature, and the maximum was observed at ca. 90 degrees C. The enzyme has a molecular mass of about 220 kDa and consists of six subunits with identical molecular masses of 36 kDa. The enzyme required NAD as a coenzyme for L-glutamate deamination and was different from the NADP-dependent glutamate dehydrogenase from other hyperthermophiles. The Km values for NAD, L-glutamate, NADH, 2-oxoglutarate, and ammonia were 0.025, 0.17, 0.0050, 0.066, and 9.7 mM, respectively. The enzyme activity was significantly increased by the addition of denaturants such as guanidine hydrochloride and some water-miscible organic solvents such as acetonitrile and tetrahydrofuran. When fluorescence of the enzyme was measured in the presence of guanidine hydrochloride, a significant emission spectrum change and a shift in the maximum were observed but not in the presence of urea. These results indicate that this hyperthermophilic enzyme may have great potential in applications to biosensor and bioreactor processes.
Amino Acid Sequence↗
A novel glycerol kinase from Flavobacterium meningosepticum: characterization, gene cloning and primary structure.
A thermostable glycerol kinase (FGK) was purified 34-fold to homogeneity from Flavobacterium meningosepticum. The molecular masses of the enzyme were 200 kDa by gel filtration and 50 kDa by SDS-PAGE. The Km for glycerol and ATP were 0.088 and 0.030 mM, respectively. The enzyme was stable at 65 degrees C for 10 min and at 37 degrees C for two weeks. The enzyme gene was cloned into Escherichia coli and its complete DNA was sequenced. The FGK gene consists of an open reading frame of 1494-bp encoding a protein of 498 amino acids. The deduced amino acid sequence of the gene had 40-60% similarity to those of glycerol kinases from other origins and the amino acid sequence of the putative active site residue reported for E. coli GK is identical to the corresponding sequence of FGK except for one amino acid residue.
Amino Acid Sequence↗
Monitoring of inhibitors of enzymatic amplification in polymerase chain reaction and evaluation of efficacy of RNA extraction for the detection of hepatitis C virus using the internal control.
Inhibitors of enzymatic amplification in serum may cause false-negative results for direct detection of hepatitis C virus (HCV) by polymerase chain reaction (PCR). This study was undertaken to demonstrate the importance of the internal control in a PCR assay for detection of HCV-RNA to monitor false-negatives due to inhibitors. HCV-RNA was extracted using RNA extraction kit (SepaGene RV-R, Sanko Junyaku) and a prototype instrument for automated specific capture of HCV-RNA with probes and magnetic bead/fluid separation (Roche Molecular Systems). The extracted HCV-RNA and internal control were detected by an automated PCR machine (Cobas Amplicor, Roche Diagnostic Systems). Addition of hemoglobin (up to 4.5 g/l) to the sera followed by RNA extraction with SepaGene RV-R had no inhibitory effect on the detection of either HCV-RNA or the internal control. In contrast, addition of heparin to the sera showed an inhibitory effect with a dose-dependent manner on the detection of both HCV-RNA and the internal control, with a greater effect at lower copy number of HCV. When HCV-RNA was extracted by the automated system, the inhibitory effect of heparin was successfully eliminated. In the assays of 65 serum samples positive for anti-HCV antibodies, positivity for the internal control indicated efficient amplification and validated 14 negative and two equivocal results for detection of HCV-RNA. Detection of the internal control was negatively correlated with viral copy number in sera suggesting competitive inhibition of high viral copy number on amplification of the internal control. Extraction, co-amplification and detection of the internal control appears useful for estimating effects of inhibitors on amplification in each assay for the detection of HCV-RNA, and for evaluating efficacy of RNA extraction methods.
Enzyme Inhibitors↗
Prevalence of "orphan" parvovirus infections in mice and rats.
"Orphan" parvovirus (OPV) infection in laboratory mice and rats was serologically surveyed for 465 mouse sera and 271 rat sera collected from 1986 to 1987 and from 1993 to 1996 in Japan. The results suggest that parvovirus infection is rare in mice but common in rats (positive rate: 13-22%) and that most putative viruses were OPVs. OPV is therefore considered to already have been harbored for at least ten years in Japan.
Animals↗
[Pulmonary function at the early stage after bone marrow transplantation].
Twenty-one patients, aged 16-41 years, were prospectively followed for pulmonary function prior to and up to nine months after bone marrow transplantation (BMT). All patients had normal radiographs and no clinical signs of pulmonary disease. The FEV1.0%, V25, and of V50/V25 were within the normal ranges for all patients. Percent vital capacity (%VC) and percent diffusing capacity of lung for CO (% DLco) dropped progressively in most of the patients up to 9 months following BMT. Pulmonary function decreased earlier in patients with GVHD as well as patients who received total body irradiation either at a high dose rate (10 cGy/min) or a high lung dose (12 Gy). These asymptomatic declines in pulmonary function were measured by frequent tests of lung function.
Adolescent↗
Stability of tricyclic antidepressants in formalin solutions.
We investigated the stability of the secondary amines, desipramine (DP) and nortriptyline (NRT), and the tertiary amines, imipramine (IP) and amitriptyline (AT), in formaldehyde (F) and paraformaldehyde (PF) aqueous solutions. NRT showed little instability in 0.37 to 37% F and PF solutions, but AT formation was detected and increased, up to 0.46 to 2.7%, in parallel with rising F and PF concentrations. DP was unstable and levels decreased to 74 to 96% with increasing F concentrations, and fell only to 96% in 10% PF solution. IP formation increased in the same manner as AT to 2.9 to 3.5% of the initial DP. When AT and IP were stored in F and PF solutions, concentrations of AT and IP did not change. DP in F pH 3 to 11 phosphate buffer (PB) solutions showed high recovery in the order: pH 5 > pH 7 > pH 9 > pH 3 and pH 11. DP in PF buffered solutions decreased slightly only at pH 3 (3.5%). By contrast, IP did not change at any pH (pH 3 to 11) of the F or PF solutions. During storage for 21 days at room temperature in 3.7% F and PF solutions, IP and DP degradation was accelerated when compared with the values in pH 3 and 7 PB solutions. However, IP detected in DP F or PF solution was only 0.2% of the initial DP 21 days after storage. Thus, AT, NRT, IP and DP degraded gradually in F and PF solutions during storage at room temperature. TCAs may first react nucleophilically with formaldehyde to form hemiaminals. DP in 3.7% formaldehyde aqueous solution formed little of its methylated product, IP, at room temperature.
Amitriptyline↗
A transferrin D variant (DHR5) identical with DSaga by polyacrylamide gel isoelectric focusing was found in a disputed paternity case.
A transferrin (TF) variant was found in the child in a paternity case by polyacrylamide gel isoelectric focusing (PAGIF). Further analysis of serum TF disclosed that this rare TF variant was inherited from the alleged father. This TF variant was identified as TF DHR5, being identical with a variant TF DSaga, in a direct comparison with several known TF D variants by PAGIF. The isoelectric points (pIs) of holo-type (Fe(II) binding type) of TF DHR5 (= DSaga) and TF C were 5.6 and 5.4, respectively. After sialidase treatment, TF DHR5 (= DSaga) showed a single band migrating at a more basic position than TF C, thus suggesting that the pI difference between the two was due to amino acid substitution.
Acrylic Resins↗
Involvement of macrophage migration inhibitory factor (MIF) in the mechanism of tumor cell growth.
BACKGROUND: Macrophage migration inhibitory factor (MIF) was recently rediscovered as a cytokine, pituitary hormone, and glucocorticoid-induced immunomodulator. MIF is constitutively expressed in various cells and enhances production of inflammatory cytokines such as tumor necrosis factor-alpha, interleukin-1, and interferon gamma. Recently, it was reported that MIF mRNA was overexpressed in prostatic tumors, which suggests that MIF is a protein involved in tumor cell growth beyond inflammatory and immune responses. MATERIALS AND METHODS: We examined the expression of MIF in the murine colon carcinoma cell line colon 26 by Western and Northern blot analyses and immunohistochemistry. Next, we investigated the effects of transforming growth factor (TGF) beta, basic fibroblast growth factor (b-FGF), and platelet-derived growth factor (PDGF) on the expression of MIF mRNA. Furthermore, we examined whether MIF is involved in tumor cell proliferation, using an MIF anti-sense plasmid transfection technique. RESULTS: We demonstrated that MIF protein and its mRNA were highly expressed in colon 26 cells, using Western and Northern blot analyses, respectively. By immunohistochemical analysis, we found that MIF was localized largely in the cytoplasm of the tumor cells. In response to TGF-beta, b-FGF, and PDGF, MIF mRNA expression was significantly up-regulated. Following this, we transfected the cells with an anti-sense MIF plasmid, which revealed that this treatment induced significant suppression of cell proliferation. CONCLUSION: Although MIF plays multifunctional roles in a broad spectrum of pathophysiological states, little has been done to investigate the role of this protein in association with tumor growth. The current results suggest the possibility that MIF induces tumor cell growth in concert with other growth factors, which encouraged us to investigate a novel approach for tumor therapy using an anti-MIF antibody and an MIF anti-sense plasmid transfection technique.
Animals↗
Muscarinic receptor-mediated dual regulation of ADP-ribosyl cyclase in NG108-15 neuronal cell membranes.
Cyclic ADP-ribose (cADP-ribose) is an endogenous modulator of ryanodine-sensitive Ca2+ release channels. An unsolved question is whether or not cADP-ribose mediates intracellular signals from hormone or neurotransmitter receptors. The first step in this study was to develop a TLC method to measure ADP-ribosyl cyclase, by which conversion of [3H]NAD+ to [3H]cADP-ribose was confirmed in COS-7 cells overexpressing human CD38. A membrane fraction of NG108-15 neuroblastoma x glioma hybrid cells possessed ADP-ribosyl cyclase activity measured by TLC. Carbamylcholine increased this activity by 2.6-fold in NG108-15 cells overexpressing m1 or m3 muscarinic acetylcholine receptors (mAChRs), but inhibited it by 30-52% in cells expressing m2 and/or m4 mAChRs. Both of these effects were mimicked by GTP. Pretreatment of cells with cholera toxin blocked the activation, whereas pertussis toxin blocked the inhibition. Application of carbamylcholine caused significant decreases in NAD+ concentrations in untreated m1-transformed NG108-15 cells, but an increase in cholera toxin-treated cells. These results suggest that mAChRs couple to ADP-ribosyl cyclase within cell membranes via trimeric G proteins and can thereby control cellular function by regulating cADP-ribose formation.
ADP-ribosyl Cyclase↗
Postmortem alcohol analysis of the synovial fluid and its availability in medico-legal practices.
Alcohol analyses using the synovial fluid of the knee joint (synovial alcohol concentration, SAC) as well as blood (BAC) and urine (UAC) were performed by the pulse heating-gas chromatographic method in 12 medico-legal autopsy cases. In 11 of the 12 cases, the regression analysis showed that BAC and UAC had a fairly linear relationship with SAC, and the average ratios of the BAC/SAC and UAC/SAC were 0.76 +/- 0.12 (range 0.60-0.94) and 1.03 +/- 0.11 (range 0.90-1.21), respectively. The measured BACs showed a difference of 11.8 +/- 9.4% (0.9-27.1%) when compared with the calculated BACs by means of the average BAC/SAC ratio. Between the measured UACs and the calculated UACs, there was an average difference of 7.9 +/- 6.0% (0.3-17.8%). The present study demonstrates that the synovial fluid is one of the available biological specimens for the prediction of BAC or UAC within a range in autopsy cases in which suitable blood and/or urine specimens cannot be obtained.
Adult↗
4-alpha-glucanotransferase from the hyperthermophilic archaeon Thermococcus litoralis--enzyme purification and characterization, and gene cloning, sequencing and expression in Escherichia coli.
4-Alpha-Glucanotransferase was purified from cells of Thermococcus litoralis, a hyperthermophilic archaeon. The molecular mass of the enzyme was estimated to be approximately 87 kDa by gel filtration. The optimal temperature for its activity was 90 degrees C. The enzyme catalyzed the transglycosylation of maltooligosaccharides, yielding maltooligosaccharides of various lengths and glucose. When maltoheptaose was used as the substrate, glucoamylase-resistant and glucoamylase-sensitive saccharides were produced. On incubation of amylose with the T. litoralis enzyme, glucoamylase-resistant but alpha-amylase-sensitive molecules were produced, but the amount of reducing sugar showed only slight increases. These results indicate that the T. litoralis enzyme catalyzes not only intermolecular transglycosylation to produce linear alpha-1,4-glucan, but also intramolecular transglycosylation to produce cyclic alpha-1,4-glucan (cycloamylose), similarly to potato 4-alpha-glucanotransferase (called disproportionating enzyme). The gene encoding the T. litoralis 4-alpha-glucanotransferase was cloned, sequenced and expressed in Escherichia coli. The nucleotide sequence of the gene encoded a 659-amino acid protein with a calculated molecular mass of 77,883 Da. The amino acid sequence of the T. litoralis enzyme showed high similarity with those of alpha-amylases of Pyrococcus furiosus, a hyperthermophilic archaeon, and Dictyoglomus thermophilum, an extremely thermophilic bacterium, but little similarity with those of other known 4-alpha-glucanotransferases.
Amino Acid Sequence↗
alpha-Galactosidase A deficient mice: a model of Fabry disease.
Fabry disease is an X-linked inherited metabolic disorder that is caused by a deficiency of alpha-galactosidase A (alpha-Gal A). Progressive deposition of neutral glycosphingolipids that have terminal a-linked galactosyl moieties in vascular endothelial cells causes renal failure along with premature myocardial infarctions and strokes in patients with this condition. No specific treatment is available for patients with this disorder at this time. An animal model of this condition would be valuable for exploring therapeutic strategies for patients with Fabry disease. We report here the generation of alpha-Gal A deficient mice by gene targeting and an analysis of the resulting phenotype. The knockout mice display a complete lack of alpha-Gal A activity. The mice, however, appeared clinically normal at 10 weeks of age. Ultrastructural analysis revealed concentric lamellar inclusions in the kidneys, and confocal microscopy using a fluorescent-labeled lectin specific for alpha-D-galactosyl residues showed accumulation of substrate in the kidneys as well as in cultured fibroblasts. Lipid analysis revealed a marked accumulation of ceramidetrihexoside in the liver and the kidneys. These findings indicate the similarity of the pathophysiological process in the mutant mice and in patients with Fabry disease. The deficiency of alpha-Gal A activity and the accumulation of material containing terminal alpha-galactosyl residues in cultured embryonic fibroblasts derived from alpha-Gal A(-/0) mice were corrected by transducing these cells with bicistronic multidrug resistance retroviruses containing human alpha-Gal A cDNA.
Animals↗
Hemangiopericytoma of the sigmoid mesentery: report of a case with immunohistochemical findings.
Hemangiopericytoma has been described in various sites in the body but only rarely in the mesocolon. This report describes the clinical course of an 83-year-old man whose mesosigmoidal tumor (hemangiopericytoma) was resected on 11 November 1994. Immunostaining was done with the following primary antibodies: alpha-actin, vimentin, factor VIII-related antigen, chromogranin, and S-100. Staining for factor VIII-related antigen was strongly positive in the endothelial cells of the capillaries, but negative in the tumor cells. The tumor cells contained immunoreactive vimentin, but demonstrated no alpha-actin, chromogranin, or S-100. Since the operation, the patient has been disease-free for 11 months.
Actins↗