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Biomedical subjects

Y Ohkubo

Publications and source records attributed to Y Ohkubo.

At least 73 records · Page 4Linked to original sources
PubMed1995

A human monoclonal antibody to a human self-antigen, CD2 derived from human peripheral blood lymphocytes engrafted in SCID mice.

To establish human hybridoma lines, production of human immunoglobulin (Ig) and behavior of the implanted human peripheral blood lymphocytes (PBL) were characterized in severe combined immunodeficiency (SCID) mice. Human PBL from healthy donors were injected into the peritoneal cavity of SCID mice, and they were immunized with self-antigen, CD2. CD45+ cells (human PBL) migrated to lymphoid tissues in the mice as early as 4 days, accounting for more than half the lymph node cells and thymocytes. The number of cells releasing human IgG specific to the antigen increased 3.5 weeks after immunization without the usual constraint that production of the IgG, an autoantibody, is prohibited by immunological tolerance in humans. Therefore, we established several human hybridomas secreting human IgG to CD2, since splenocytes and lymph node cells from the implanted SCID mice at 3.5 weeks were fused with a human B lymphoblastoid cell line. A human anti-CD2 monoclonal antibody (MAb) was confirmed to bind to natural CD2 on human T cells by flow cytometric analysis. The epitope for the MAb was identical with a portion that the ligand LFA-3 binded, so that the MAb might reduce the inflammatory reaction caused by preventing activation of human T cells. Here, we report that the human immune system could be reconstituted in SCID mice to develop human hybridomas producing human MAb to a human self-antigen.

Animals↗
PubMed1995

Tiapride attenuates pain transmission through an indirect activation of central serotonergic mechanism.

Tiapride dose-dependently attenuated the biphasic nociceptive responses induced by s.c. injection of formalin to the hindpaw of mice, and its activity on the first (ED50 = 110 mg/kg p.o.) and the second (ED50 = 32.0 mg/kg p.o.) phases paralleled that on the nociceptive response to intrathecal injection of substance P (ED50 = 190 mg/kg p.o.) and somatostatin (ED50 = 56.0 mg/kg p.o.), respectively. Moreover, a similar antinociceptive activity was observed in streptozotocin-induced diabetic or genetically diabetic (db/db) mice. The effects of tiapride (100 mg/kg p.o.) on both phases of the formalin test in normal mice were abolished by pretreatment with p-chlorophenylalanine (800 x 2 mg/kg p.o.), a 5-hydroxytryptamine (5-HT) depletor, or pindolol (1 mg/kg i.p.), a 5-HT1 antagonist, but were scarcely affected by 3-tropanyl-indole-3-carboxylate, a 5-HT3 antagonist. Ketanserin (1 mg/kg i.p.), a 5-HT2 antagonist, attenuated the effect of tiapride on the second phase but not on the first phase. This study on the antinociceptive mechanism of action of tiapride (that blocks painful neuropathy in diabetic patients) has led us to hypothesize that the drug attenuates pain transmission through an indirect activation of central 5-HT1 and 5-HT2 receptors.

Animals↗
PubMed1995

FR121196, a potential antidementia drug, ameliorates the impaired memory of rat in the Morris water maze.

We investigated the effects of FR121196 N-(4-acetyl-1-piperazinyl)-4-fluorobenzenesulfonamide on the performance of rats in the Morris water maze using a two-trial-per-day regimen. Escape latency to find the platform was significantly prolonged in young rats subjected to scopolamine (1 mg/kg) treatment or to nucleus basalis magnocellularis (NBM) or fimbria-fornix (FF) lesioning compared with the respective control animals. Similar memory deficit was observed in aged rats (24-26 months old) compared with young rats. Daily treatment with FR121196 (0.1-1.0 mg/kg i.p.) ameliorated all the memory deficits except in the FF-lesioned rats, whereas physostigmine (0.032-0.32 mg/kg) ameliorated the memory deficits in scopolamine-treated rats but not in the NBM-lesioned, FF-lesioned or aged rats. Methamphetamine (0.32-3.2 mg/kg) hardly ameliorated the memory deficits and, rather, aggravated those in the scopolamine-treated, NBM-lesioned and aged rats. Neurochemical analysis of the brain showed that dopamine, norepinephrine and serotonin levels were significantly reduced in the hippocampus of the FF-lesioned rats compared with the sham control, and a significant reduction of dopamine level in the cortex was also observed in the aged rats compared with young rats. None of these parameters were changed after NBM lesioning. We concluded that FR121196 ameliorates the memory deficits of rats with cholinergic dysfunction through the action on the hippocampal monoaminergic (possibly serotonergic) neurons.

Age Factors↗
PubMed1994

Characterization of a Rana catesbeiana lectin-resistant mutant of leukemia P388 cells.

Sialic acid-binding lectin (SBL-C) from Rana catesbeiana eggs inhibits the growth of tumor cells such as P388 and L1210 leukemia cells (K. Nitta et al., Cancer Res., 54: 920-927, 1994). Here we report the establishment of an SBL-resistant P388 variant cell line, RC-150. Both P388 and RC-150 cells were agglutinated by SBL-C; however, growth of RC-150 cells was unaffected by SBL-C. Cytoplasmic free Ca2+ concentration and transglutaminase activity of RC-150 cells were 0.5 (110 nM) and 3 times (0.62 nmol/mg/min) as high as those of P388 cells, respectively. Microvilli and microplicae were observed on the surface of P388 cells by scanning electron microscopy but were rarely seen on RC-150 cells. Dansylcadaverine-labeled SBL-C bound to both P388 and RC-150 cells. Binding of SBL-C to these tumor cells appears to be mediated by two species of wheat germ agglutinin-stained cell membrane sialoglycoproteins. Labeled SBL-C entered P388 but not RC-150 cells, suggesting that internalized SBL-C acts as an inhibitor of cell proliferation.

Agglutination↗
PubMed1994

SV40 large T antigen reinduces the cell cycle in terminally differentiated myotubes through inducing Cdk2, Cdc2, and their partner cyclins.

Terminally differentiated skeletal muscle myotubes are arrested in G0 phase of the cell cycle and are unable to be released from this arrest by stimulation with mitogens including serum and growth factors. To inspect a possibility of reversing the quiescence at the G0 phase, we have exploited the mouse skeletal muscle cell line C2SVTts11, which is a clone of C2 cells transfected with the SV40 T antigen gene (encoding thermolabile large T and wild-type small t) fused to an inducible promoter. When the large T is induced in the myotubes, the terminally differentiated cells reenter the cell cycle and proceed to S and M phases. To elucidate how large T forces the myotubes to traverse each phase of the cell cycle, we examined the expression and activity of Cdk2 and Cdc2, which in complex with cyclin A and cyclin B are essential for S and M phases, respectively in undifferentiated cells. The levels of their mRNAs and proteins and histone H1 kinase activity, which was ascribed to Cdc2-cyclin B, were high in the proliferating myoblasts but gradually decreased during terminal differentiation. In contrast, they were reinduced in the myotubes reentering the cell cycle. Stimulation of the myotubes with serum failed to evoke these factors. These results indicate that large T, but not mitogens, is able to drive terminally differentiated myotubes to pass each phase of the cell cycle through eliciting these factors as do mitogens on proliferating undifferentiated cells. Since large T is a nuclear protein, signals generated by the protein in the nucleus are likely to be sufficient to induce each phase of the cell cycle in the terminally differentiated cells.

Animals↗
PubMed1994

Effect on insulin sensitivity of angiotensin converting enzyme inhibitors with or without a sulphydryl group: bradykinin may improve insulin resistance in dogs and humans.

The present study compared the effect on insulin sensitivity of ACE inhibitors with a sulphydryl group (captopril) or those without a sulphydryl group (delapril and enalapril) during the hyperinsulinaemic euglycaemic clamp test in both animal and clinical experiments. A possible contribution of bradykinin to the improvement of insulin sensitivity by ACE-inhibition was also studied. In healthy control and depancreatized dog experiments, administration of captopril either intravenously (3.0 mmol.kg-1) or orally (5.0 mmol.kg-1) increased insulin sensitivity indices and plasma bradykinin concentrations. In comparison, intravenous administration of an active metabolite of delapril (3.0 mmol.kg-1) and oral administration of either delapril or enalapril (5.0 mmol.kg-1) showed slight, but not significant increases in insulin sensitivity indices and plasma bradykinin concentrations. Infusion of a bradykinin antagonist (N-alpha-adamantane-acetyl-D-Arg-[Hyp3,Thi5,8,D-Phe7]-b bradykinin) (0.5 nmol.kg-1 x min-1) abolished the effect of captopril on insulin sensitivity. Furthermore, intravenous administration of bradykinin (0.1 nmol.kg-1 x min-1) increased insulin sensitivity indices. In clinical experiments, insulin sensitivity indices decreased in the following order: normotensive healthy subjects, hypertensive non-diabetic patients, normotensive NIDDM patients and hypertensive NIDDM patients. In these four groups, oral administration of captopril (2.0 mmol.kg-1) significantly increased insulin sensitivity indices, and a concomitant increase in plasma bradykinin concentrations was observed. By contrast, oral administration of enalapril or delapril showed slight, but not significant effects on insulin sensitivity indices and plasma bradykinin concentrations. From these studies, it is concluded that ACE inhibitors with a sulphydryl group have more potent action on the improvement in insulin sensitivity than those without a sulphydryl group.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1994

Concentration of 226Ra in human teeth.

226Ra concentrations in human teeth from several cities, mainly Tokyo, Japan, were determined with emphasis on the measurement of low-level 226Ra by alpha-ray spectrometry following chemical separation. No appreciable differences in 226Ra concentration were found among various permanent teeth samples of different age groups in Tokyo. The mean 226Ra concentration for Tokyo was 0.51 +/- 0.06 mBq (g Ca)-1. 226Ra concentration [mean: 0.67 +/- 0.11 mBq (g Ca)-1] in teeth in western regions of the country was statistically higher than that [mean: 0.48 +/- 0.09 mBq (g Ca)-1] in eastern ones. The mean 226Ra concentration [0.51 mBq (g Ca)-1] in teeth from Tokyo was less than the concentration [1.11 mBq (g Ca)-1] reported for vertebral bone samples of this city.

Adolescent↗
PubMed1994

Involvement of membrane-bound transglutaminase in the invagination of transferrin into rat reticulocyte plasma membrane.

We demonstrated the invagination of transferrin into reticulocyte plasma membrane to learn whether membrane-bound transglutaminase (TGase, a Ca(2+)-dependent enzyme) is involved in this invagination. The invagination was assessed by acid-resistance assay and antibody-inaccessibility assay. The invagination was blocked in the absence of ATP. [14C]Putrescine, a substrate for TGase, was incorporated into the membrane during the invagination. This incorporation was decreased in the absence of ATP or transferrin and was completely blocked in the presence of monodansylcadaverine or EGTA. The TGase inhibitor and EGTA also decreased the invagination. In the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, the labeling of 43 kDa membrane protein with [14C]putrescine and the increase in aggregation of proteins were observed in a transferrin-, ATP- and Ca(2+)-dependent manner. These results provide the first evidence for modification of protein by TGase accompanying the invagination of transferrin into the membrane, and suggest that membrane-bound TGase is involved in the invagination step of endocytosis.

Acids↗
PubMed1994

A screening concept based on a hypothesis led to the development of a putative cognitive enhancer that stimulates penile erection.

Starting from the hypothesis that drugs which specifically activate the hippocampal cholinergic nerve activity may ameliorate memory impairments, we carried out a series of evaluations for a novel cognitive enhancer using enhancement of penile erection as a sign of cholinergic activation, and found FR64822. The compound facilitated penile erection in naive rats, and it ameliorated scopolamine-induced amnesia of rats in passive avoidance tasks with bell-shaped dose-response curves, while it dose-dependently reduced body weight gain in Zucker fatty rats. Pretreatment with sulpiride (32 mg/kg, p.o.) hardly affected the former two activities, but significantly reduced the anorectic activity in Zucker rats. Further evaluation of FR64822 derivatives characterized a second compound, FR121196, which induces penile erection and memory enhancement, but not body weight reduction. Memory enhancing and erection stimulating activities of FR121196 were abolished in rats treated with either cysteamine (200 mg/kg, s.c.), a somatostatin depletor, or lesioning of the serotonergic raphe nuclei. Thus, classic whole animal studies based on a hypothesis proved to be efficient for reaching our objective, the discovery of a new drug. They also gave us insight into the common somatostatinergic and serotonergic mechanisms underlying penile erection and memory improvement.

Animals↗
PubMed1994

[Bladder carcinoma producing granulocyte colony stimulating factor (G-CSF). A case report].

A 69-year-old woman was admitted to the hospital complaining of general fatigue and lower abdominal pain. She had undergone total cystectomy because of invasive recurrent bladder carcinoma three months ago. Histopathological diagnosis was transitional cell carcinoma (TCC) grade 3 and squamous cell carcinoma (SCC), pT3a. A goose egg-sized painful mass was noticed at the lower abdominal region. A CT scan revealed an intrapelvic fist-sized mass and suggested tumor recurrence with ileus caused by intestinal invasion. The laboratory examination showed remarkable leukocytosis of 79,700/mm3 in the peripheral blood and serum analysis revealed high value of granulocyte colony stimulating factor (G-CSF), 240 pg/ml (normal: less than 30 pg/ml). In spite of active treatment, the patient died of cachexia about a month after detection of the leukocytosis. The autopsy showed that the recurrent tumor had positive immunohistochemical staining for G-CSF, and the bone marrow had reactive proliferation mainly by granulocytes. From these findings, this case was diagnosed as bladder carcinoma producting G-CSF. G-CSF producting tumor of the bladder is very rare. This was the 8th case in Japanese literatures. The previous reports were reviewed and discussed.

Aged↗
PubMed1994

[Hyperbaric oxygen therapy in the successful treatment of two cases of radiation-induced hemorrhagic cystitis].

Hemorrhagic cystitis resulting from radiation to pelvic visceral malignant lesions often might be incurable and there have been established no definitive treatment. We experienced 2 cases with radiation-induced severe hemorrhagic cystitis refractory to conventional therapy. The treatment with hyperbaric oxygen to control hematuria was performed and obtained successful results. Gross hematuria was disappeared and cystoscopic figure was remarkably improved. No remarkable side-effect was observed in both patient. This experience suggested that hyperbaric oxygen could be considered the primary treatment for patient with radiation-induced hemorrhagic cystitis instead of usual treatment.

Aged↗
PubMed1993

Dephosphorylation on tyrosine of epidermal growth factor receptor is inhibited by Ca2+ pretreatment in isolated liver membrane.

The autophosphorylation of epidermal growth factor receptor of the liver membrane was enhanced by pretreatment with 1 mM Ca2+. This enhancement was blocked by 30 microM genistein, but not by 30 microM H-7. In immunoblotting analysis, the amount of phosphotyrosine increased in the Ca(2+)-pretreated membrane. When dephosphorylation processes were studied, receptor dephosphorylation occurred in a time-dependent manner in the untreated membrane, but not in the Ca(2+)-pretreated membrane. In the presence of sodium vanadate, receptor phosphorylation increased in the untreated membrane, but not in the Ca(2+)-pretreated membrane. Inhibition of dephosphorylation did not occur in the membrane pretreated with Ca2+ in the presence of monodansylcadaverine, a transglutaminase (TGase) inhibitor. These results suggest that Ca2+ pretreatment causes the inhibition of some protein tyrosine phosphatase via a Ca(2+)-involved mechanism such as activation of TGase and results in the enhancement of receptor phosphorylation.

Adenosine Triphosphate↗
PubMed1993

Calcium pretreatment induces the decrease in epidermal growth factor binding through the activation of transglutaminase in isolated liver membrane.

Pretreatment of hepatocytes with Ca2+ and A23187 resulted in the decrease in epidermal growth factor (EGF) binding to the receptor. This decrease could be induced by only the pretreatment of the isolated liver membrane with Ca2+. By the pretreatment with 1 mM Ca2+ for 30 min, EGF binding was decreased to 40% of control. Scatchard plot analyses indicated that high affinity binding sites of EGF receptor disappeared after Ca2+ pretreatment. No effect of Ca2+ pretreatment on the degradation of receptor occurred. The Ca2+ effect was prevented by monodansylcadaverine or iodoacetamide, transglutaminase (TGase) inhibitors. After membrane-bound TGase was inactivated, the Ca2+ effect did not occur, except in the presence of purified TGase. Labeled putrescine was incorporated into some proteins of membrane with Ca2+ treatment. These results suggest that Ca(2+)-activated TGase induces the decrease in EGF binding to the receptor via modification of some membrane protein.

Animals↗
PubMed1993

[New microwave transurethral hyperthermia for benign prostatic hyperplasia--clinical evaluation].

Several types of hyperthermic apparatuses are employed to treat benign prostatic hyperplasia (BPH). As the prostate surrounds the urethra, we believed that transurethral heating allowed for more efficient and uniform heating. A new effective apparatus for this objective was developed. The size of our hyperthermic apparatus was about 52 x 45 x 20 centimeters and the originating frequency was 2,450 +/- 30 MHz. We used T-type thermocouples as temperature sensors. The transurethral applicator had a cooling system. Thirty patients complaining of obstructive symptoms due to BPH were treated. Hyperthermia was performed 3-6 times for each patients (two times per week). Each procedure was performed for 60 minutes. The temperature was controlled at 39 degrees C on the urethral surface (43 degrees C at prostate). The overall efficacy of this treatment was effective in 23 of the 30 patients (76.7%). In addition, there were no severe complications. As the size of this apparatus was miniaturized, it could be used at the bed side.

Aged↗
PubMed1992

Effect of retinoic acid on liver transglutaminase activity and carbon tetrachloride-induced liver damage in mice.

Transglutaminase (TGase) activity in the cytosol fraction of the mouse liver increased following intraperitoneal injection of retinoic acid. Retinoic acid inhibited the carbon tetrachloride-induced increase in serum alanine transaminase activity. These findings suggest that TGase is involved in the effect of retinoic acid on carbon tetrachloride-induced liver damage.

Alanine Transaminase↗
PubMed1992

Application of 67Ga for the estimation of reticulocyte production.

In order to estimate the production of reticulocytes, which have a larger number of transferrin receptors than erythrocytes, we used 67Ga which is exclusively bound to transferrin in the blood. The pattern of uptake of 67Ga by reticulocytes was quite similar to the time course of transglutaminase activity which might be involved in receptor-mediated endocytosis. The preinjection of Fe3+ decreased the uptake of 67Ga by reticulocytes. These results suggested that 67Ga in a transferrin-bound form was taken up by reticulocytes via receptor-mediated endocytosis. It was showed that the application of 67Ga is very easy and useful for the estimation of reticulocyte production.

Animals↗
PubMed1992

Type I hyperlipoproteinemia caused by lipoprotein lipase defect in lipid-interface recognition was relieved by administration of medium-chain triglyceride.

We have previously reported lipoprotein lipase with a defect of lipid-interface recognition in a patient with type I hyperlipoproteinemia. In this patient, lipoprotein lipase from post-heparin plasma (PHP) hydrolyzed monomeric substrate tributyrin, but scarcely hydrolyzed triolein emulsified with Triton X-100 and that in very-low-density lipoproteins ([VLDL] d < 1.006 g/mL), and did not bind to VLDL. The triglyceride (TG) level of this patient did not decrease to less than 1,000 mg/dL with a low-fat diet (1,400 kcal containing 10 g fat/d). When the patient took 30 g medium-chain TG (MCT) in addition to the 1,400-kcal diet, her serum TG level decreased to 250 mg/dL and her clinical signs improved. The low clearance rate of serum TG with heparin injection improved after intake of MCT. Caproic acid levels were maintained at 1.4% and 2.6% in chylomicrons and VLDL after MCT intake, respectively. The patient's lipoprotein lipase hydrolyzed triolein emulsified with 2% tricaprin at the same rate as that of control lipoprotein lipase. The patient's lipoprotein lipase-catalyzed hydrolyzing rate of triolein in chylomicrons obtained after MCT administration was also enhanced up to 70% of that of control lipoprotein lipase. These findings suggest that hypertriglyceridemia caused by lipoprotein lipase with a defect in lipid-interface recognition could be relieved with the administration of medium-chain TG, and that one of the mechanisms of this effect might be a modification of TG-rich lipoproteins by MCT.

Adolescent↗