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Biomedical subjects

T Ohno

Publications and source records attributed to T Ohno.

At least 451 records · Page 25Linked to original sources

Enhancement by protein kinase C of prostacyclin receptor-mediated activation of adenylate cyclase through a calmodulin/myristoylated alanine-rich C kinase substrate (MARCKS) system in IC2 mast cells.

The addition of 12-O-tetradecanoylphorbol-13-acetate (TPA) markedly enhanced cAMP formation induced by carbacyclin, a stable prostacyclin analogue, in cultured mast cells (IC2 cells), but did not enhance basal or NaF plus AlCl3-induced cAMP formation. On the other hand, W-7, a calmodulin (CaM) inhibitor, almost completely suppressed the enhancing activity of TPA, suggesting the involvement of CaM in the enhancement by TPA of carbacyclin-induced cAMP formation. The enhancing activity of TPA disappeared in TPA-treated cells permeabilized with saponin in the presence of Ca2+, but reconstitution with CaM in the permeable cells resulted in remarkable restoration of the action of TPA. On the other hand, TPA treatment induced the phosphorylation and translocation of myristoylated alanine-rich C kinase substrate (MARCKS) from the membrane to the cytosol. Exogenously added protein kinase C (PKC) also phosphorylated MARCKS and induced its translocation in the cells permeabilized with saponin. Whereas the addition of CaM did not enhance the carbacyclin-stimulated GTPase activity and adenylate cyclase activity in the control permeable cells, in which MARCKS bound to the membrane, CaM markedly enhanced those activities in the PKC-treated permeable cells, which lost endogenous membrane-bound MARCKS. When MARCKS was added to the PKC-treated permeable cells, MARCKS bound to the membrane and inhibited the effects of CaM. These results suggest that activation of PKC enhances the prostacyclin-activated adenylate cyclase through a CaM/MARCKS system.

Adenylyl Cyclases↗

Plasminogen activators and their inhibitors in leukemic cell homogenates.

Plasminogen activator (PA) and PA inhibitor (PAI) were measured in homogenates of leukemia cells. Both PA and PAI levels were higher in non-lymphoblastic leukemia than in lymphoblastic leukemia. The levels were below the sensitivity of determination in chronic myelocytic leukemia (CML) but showed significant increases in blast crisis (CML,bc). The level of the tissue type PA (t-PA) antigen was highest in acute myeloblastic leukemia (AML) and that of the urokinase type PA (u-PA) was highest in acute promyelocytic leukemia (APL). The PAI-I antigen showed no marked cell specificity, but the PAI-II antigen was markedly increased in myelomonocytic leukemia and acute monocytic leukemia (AMoL). From these findings, various PAs and PAIs are considered to be present in leukemia cells and to be involved in hemostatic disorders, thus they are of diagnostic value in leukemia.

Blood Cells↗

Ubiquitous nuclear proteins bind to 5' upstream region of major Kunitz chymotrypsin inhibitor gene in winged bean.

Winged bean Kunitz chymotrypsin inhibitor (WCI) accumulates abundantly in seeds and tuberous roots of winged bean plant. In seeds, the WCI mRNA is observed transiently during seed maturation period. The WCI is encoded by a multigene family and the major WCI (WCI-3) is encoded by two nearly identical genes (WCI-3a and WCI-3b genes), in which nucleotide sequences in the 1.1 kb 5' flanking regions are about 99% homologous to each other and the transcribed regions are completely identical. Here we report the detection of two types of nuclear proteins which bind to the multiple sites in the 5' upstream region of the WCI-3a gene. One of the proteins, band 1-forming protein, also bound to cauliflower mosaic virus 35S (CaMV35S) promoter, but another protein, band 3-forming protein, did not. DNaseI footprinting analysis showed that these proteins bound to AT-rich upstream regions in the WCI-3a gene. Addition of poly(dA-dT)-poly(dA-dT) to the binding reaction inhibited the formation of the retarded bands, while poly(dI-dC)-poly(dI-dC) did not. In various organs and throughout seed maturation period, proteins with invariable binding specificities were detected, and these binding proteins met some operational criteria for high-mobility-group (HMG) proteins. These results suggest that leguminous seed AT-binding proteins reported on several seed storage protein genes may be HMG-like proteins which are present ubiquitously in plant organs.

Base Sequence↗

Detection of a cryptic paracentric inversion within band 11p13 in familial aniridia by fluorescence in situ hybridization.

We report the first familial case of dominantly inherited aniridia with a cryptic inversion within band 11p13. High-resolution chromosome analysis gave a suspicion of a tiny constitutional aberration around band 11p13 and fluorescence in situ hybridization using 11p cosmids successfully confirmed that the aniridia patients of this family have an inversion within band 11p13. The distal breakpoint of the inversion is telomeric to a candidate aniridia gene (AN2) and suggests that more genes might be involved in the etiology of aniridia. In situ hybridization is a powerful tool to detect cryptic rearrangements in sporadic or familial patients with aniridia. This family indicated the importance of careful observation of the 11p13 region of aniridia patients, even if the aniridia was autosomal dominantly inherited.

Adult↗

Presence of gonadotropin-releasing hormone and its messenger ribonucleic acid in human ovarian epithelial carcinoma.

OBJECTIVE: The purpose of this study was to investigate the expression of gonadotropin-releasing hormone messenger ribonucleic acid and the presence of gonadotropin-releasing hormone in human ovarian carcinoma known to have gonadotropin-releasing hormone binding sites and to be affected by gonadotropin-releasing hormone analog. STUDY DESIGN: Human ovarian carcinomas surgically removed and human ovarian carcinoma cell lines were examined. Gonadotropin-releasing hormone was determined by a radioimmunoassay and a bioassay. Gonadotropin-releasing hormone messenger ribonucleic acid was determined by reverse transcription polymerase chain reaction using oligonucleotide primers synthesized according to the published human gonadotropin-releasing hormone sequence. RESULTS: Gonadotropin-releasing hormone was shown to be present in extracts of ovarian mucinous cystadenocarcinoma sample (0.8 +/- 0.12 pg/mg of protein) and ovarian adenocarcinoma cell line SK-OV3 (0.92 +/- 0.17 pg/mg of protein) but not in the normal ovary and placenta. Two of two extract samples from individual cases evoked dose-dependent phosphoinositide breakdown in rat granulosa cells similar to that caused by authentic gonadotropin-releasing hormone. Gonadotropin-releasing hormone messenger ribonucleic acid was detected in two of two mucinous cystadenocarcinoma specimens, one of one serous cystadenocarcinoma, and SK-OV3 cells but not in the dysgerminoma, mucinous cystadenoma, and normal ovary and placenta. CONCLUSION: The demonstration of gonadotropin-releasing hormone and its messenger ribonucleic acid raises the possibility that gonadotropin-releasing hormone may play an autocrine regulatory role in the growth of ovarian carcinoma.

Adenocarcinoma↗

Morphogenesis and origin of fibrous long-spacing collagen fibers in collagenase-treated mouse skin tissues.

Morphogenesis and origin of fibrous long-spacing collagen (FLS) fibers in newborn mouse skin tissues treated with collagenase were examined using ultrastructural observation, morphometry, histochemical methods, and immunoelectron microscopy. The enzyme caused both the partial destruction of basal laminae and the formation of abundant FLS fibers in the dermal matrix. The fibers were usually distributed in the vicinity of basal laminae in the capillaries or basal layer cells. The fibers were characterized by the cross-striated dark bands with about 91 nm periodicity and longitudinally aligned filaments with a diameter of about 6.5 nm. The dark bands of FLS fibers were often continuous with the basal laminae. Histochemical results showed that the dark bands contained the similar mucopolysaccharides which were involved in the basal laminae. Immunoelectron microscopic results showed that laminin was present in the dark bands as well as in the basal laminae, and that type VI collagen was located in the filaments of FLS fibers. These results suggest that the dark bands are formed by products similar to basal laminae and that the products were precipitated on type VI collagen-contained filaments with periodic intervals of about 91 nm. Morphometric examination revealed that there was no differences in ultrastructure between FLS fibers of a collagenase-treated mouse and those of a human neural tumor.

Animals↗

Incubation-derived calcium carbonate equivalence of papermill boiler ashes derived from sludge and wood sources.

The burning of a papermill sludge and wood mixture and landspreading the resulting ash is a potential means of disposal of papermill sludge without the use of valuable landfill space. This study evaluated the effectiveness of ashes derived from a mixture of papermill sludge and wood sources to act as an alternative liming agent. The calcium carbonate equivalence of the material was determined using a 91-day laboratory incubation test with three mineral soils and one organic horizon soil. Application rates of soil-incorporated sludge-ash ranged from 2.30 to 32.2 g per kg soil. Soil pH increased linearly with increasing sludge-ash application rate. The calcium carbonate equivalence of the material varied temporally and the average value ranged from 19% to 28%. The fraction of total P, K and Mg added with the sludge-ash and extracted from the ash-amended soils using an NH4OAc based soil test method were 2.6, 3.8 and 17.6%, respectively. The low soil test extractability of ash-derived plant nutrients suggests that this material would provide only a modest increase in plant available nutrient levels in landspread fields.

Journal Article↗

Sodium retention and hepatic function following partial portal vein ligation in the rat.

Recent studies have suggested that the development of sodium retention in experimental cirrhosis in the rat occurs when hepatic function, measured by the aminopyrine breath test, decreases below a critical threshold. The present study evaluated the relationship between renal sodium handling and hepatic function and determined whether sodium retention occurs following partial portal vein ligation. Sodium balance, urine volume, creatinine clearance and the aminopyrine rate constant of elimination, on a constant sodium intake, were evaluated daily, from 1 day before surgery to 5 days after surgery, in both sham-operated (n = 6) and partially portal vein-ligated rats (n = 14). In the partially portal vein-ligated group, sodium retention occurred in 9 rats between 1 and 4 days after surgery, accompanied by a 45% reduction in the aminopyrine rate constant of elimination. Spontaneous natriuresis occurred within 5 days after surgery, and was associated with an increase in the aminopyrine rate constant of elimination from 0.94 +/- 0.07 x 10(-2) min-1 on the last day of sodium retention to 1.36 +/- 0.06 x 10(-2) min-1 on the day of diuresis (P < 0.05). In contrast, creatinine clearance did not change throughout the study. There was a negative curvilinear association between sodium balance and the aminopyrine rate constant of elimination (r = 0.70, P < 0.001). In the five rats without sodium retention, there was no change in the aminopyrine rate constant of elimination and creatinine clearance over the 5 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Aminopyrine↗

Empathy and aggression: effects of self-disclosure and fearful appeal.

On the basis of the familiarity-empathy assumption that self-disclosure evokes empathy for the speaker, it was predicted that a victim's self-disclosure would inhibit aggression against the victim. Female Japanese subjects were asked to give electric shocks to a female victim who disclosed information about herself, was not given an opportunity to do so, or rejected disclosure. Independently of self-disclosure, another empathy arousal was introduced, that is, whether or not the victim expressed her fear of shocks before they were delivered. Consistent with our hypothesis, subjects selected less severe shocks when the victim disclosed information about herself than when she was not given an opportunity to do so or when she rejected self-disclosure. The victim's expression of fear was also very effective in reducing subjects' aggression, suggesting that drawing subjects' attention to the victim's negative emotional state evoked empathy for her and reduced their aggression.

Aggression↗

Genistein augments cyclic adenosine 3'5'-monophosphate(cAMP) accumulation and insulin release in MIN6 cells.

Effects of genistein on insulin release were studied using MIN6 cells, a glucose-sensitive insulinoma cell line. At the non-stimulatory concentrations of glucose, genistein did not affect insulin release, however, at the stimulatory concentrations of glucose, genistein significantly increased insulin release in a dose-dependent manner up to 20 micrograms/ml. The content of cAMP in MIN6 cells was also elevated significantly by genistein and the dose-response relationship between the genistein and cAMP accumulation was consistent with the relationship between the genistein and insulin release. These effects were inhibited by calcium antagonists or by the omission of extracellular calcium. Isobutylmethylxanthine (IBMX;0.1mM) increased both cAMP accumulation and insulin release in MIN6 cells and there were no additive effects by the addition of genistein. The accumulation of cAMP might have, at least in part, resulted from phosphodiesterase inhibition by genistein. These results suggest that genistein augments glucose-induced insulin release by the contribution of cAMP accumulation and calcium modulation which depends on extracellular calcium.

1-Methyl-3-isobutylxanthine↗

Complement-dependent virolysis of HIV-1 with monoclonal antibody NM-01.

Previous studies have described the isolation of monoclonal antibodies that neutralize human immunodeficiency virus type 1 (HIV-1). However, although certain antibodies are capable of blocking infection of CD4+ cells, the antibody-HIV-1 complex could potentially remain infectious and enter cells by an Fc receptor-mediated mechanism. Indeed, one goal of HIV-1 therapy with antibodies is the development of reagents capable of killing virus. The present studies with murine monoclonal antibody (MoAb) NM-01 demonstrate complement-mediated lysis of HIV-1. NM-01-dependent virolysis has been visualized by electron microscopy. Virus particles treated with NM-01 and rabbit or human complement undergo a process of outer envelope rupture and loss of the electron-dense core. The involvement of human complement in this process was confirmed by the presence of C9 in association with MoAb NM-01 immune complexes. In contrast, virolysis was not detectable with antibody or complement alone. Moreover, although MoAb NM-01 alone is capable of neutralizing HIV-1, the presence of rabbit complement was associated with over a 10-fold decrease in infectivity. Similar but less pronounced effects were observed with human complement. These findings support a potential role for antibody-dependent complement-mediated virolysis in HIV-1 therapy.

Amino Acid Sequence↗

Antiarrhythmic agents act differently on the activation phase of the ACh-response in guinea-pig atrial myocytes.

1. Anti-acetylcholine effects of pilsicainide, flecainide, disopyramide and propafenone on the acetylcholine (ACh)-induced K+ current (IK.ACh) were examined in dissociated guinea-pig atrial myocytes under whole-cell voltage clamp by the use of the 'concentration-clamp' technique. 2. The IK.ACh was activated with a latency of about 100 ms after 1 microM ACh application and desensitized to a steady-state level. The latent period and the time to peak response were shortened with increasing ACh concentration. 3. The values of half-maximal inhibition (IC50) on the peak and steady state responses were 25 and 25 microM for pilsicainide, 1.7 and 2.0 microM for disopyramide, 19 and 2.0 microM for flecainide and 0.7 and 0.2 microM for propafenone, respectively. 4. Pilsicainide and disopyramide increased the latent period and the time to peak of IK.ACh in a concentration-dependent manner. Flecainide and propafenone did not change the latent period, but shortened the time to peak and hastened the decay of IK.ACh in a voltage-independent manner. 5. The results suggest that the mechanisms underlying the anti-acetylcholine effect of antiarrhythmic drugs are different among these drugs: i.e., pilsicainide and disopyramide mainly block the muscarinic ACh receptors while flecainide and propafenone inhibit the K+ channel itself as open channel blockers.

Acetylcholine↗

Flavin-photosensitized monomerization of dimethylthymine cyclobutane dimer in the presence of magnesium perchlorate.

We have investigated the photosensitized monomerization of the cis,syn-cyclobutane dimer of 1,3-dimethylthymine using riboflavin tetraacetate and a 5-deazaflavin derivative as photosensitizer. Although little monomerization of the dimer is induced by photoexcitation of the flavins in the absence of any additives, the flavins can function as an efficient photosensitizer in the presence of magnesium perchlorate. Mechanistic studies involving spectroscopic, quantum-yield and flash-photolysis measurements demonstrated that the photosensitized monomerization exclusively proceeds through electron transfer from the dimer to the triplet flavins complexed with Mg2+. The effects of magnesium perchlorate are compared with those on the chloranil-photosensitized monomerization and also with the effects of HClO4 on the flavin-photosensitized reaction.

Flavins↗