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M Ono

Publications and source records attributed to M Ono.

At least 433 records · Page 24Linked to original sources
PubMed1995

A simple method based on PCR for detecting the relative mRNA amounts of the four mouse IgG subclasses.

A simple method based on the polymerase chain reaction (PCR) was developed for detecting the relative mRNA amounts of the four mouse IgG subclasses in total RNA samples and is described in this report. The main features of this method are, first, cDNA amplification including VH through the constant region(CH2 domain) of each IgG subclass with a set of consensus PCR primers(VH1BACK and 32P-labeled C gamma 32), and secondly, cleavage of the amplified DNA fragments with BamHI and XhoI endonucleases which act at distinct cleavage sites in the constant region of each IgG subclass. The radioactive intensities of the different sized fragments separated on polyacrylamide gel were used to show the relative amounts of IgG subclasses at the RNA level. This method provides clear detection of each IgG subclass using RNA samples from tissues in which Ig-producing cells are rare.

Animals↗
PubMed1995

Demonstration of cells possessing tolerance-inducing activity in Xenopus laevis rendered tolerant perimetamorphically.

J-strain (JJ) Xenopus laevis is easily made tolerant to semixenogeneic (X laevis x X borealis: JB) adult skin grafted onto immunologically competent larvae at stages 53-54. If the larvae are thymectomized shortly before skin grafting (late Txd), tolerance will never be induced. This suggests that suppression of the immune response to JB skin is mediated by a population of thymus-derived cells. In an attempt to confirm the presence of these cells, the spleen and tolerated JB skin were tested for their tolerance-inducing activity, by grafting either spleen or skin and a new JB skin piece simultaneously to late Txd JJ larva (secondary host). The results clearly indicated that both spleen and skin possessed the ability to induce tolerance. The injection of 1 x 10(5) splenocytes resulted in induction of tolerance in more than 80% of secondary hosts, and the rate of tolerance induction increased in relation to the injected cell number. Furthermore, tolerance was induced in about 90% of the animals by injecting as few as 100 cells isolated from the tolerated skin. Immunohistochemical observation of grafted skin showed that many host-derived T cells were distributed around the epidermal basal lamina. These results indicate that the cells with suppressive activity are a population of T cells that have differentiated in metamorphosing thymuses and have been released peripherally.

Animals↗
PubMed1995

DNase I-hypersensitive sites in the chromatin of rat growth hormone gene locus and enhancer activity of regions with these sites.

In this study, a determination was made of the chromatin structure of the rat growth hormone (GH) gene locus by DNase I sensitivity analysis using GC [GH+, prolactin (PRL)-], 235 (GH-, PRL+), GH3 (GH+, PRL+) and liver (GH-, PRL-) cells. From 7 kb upstream from the transcription start site to 19 kb downstream from the polyadenylation site, two major DNase I-hypersensitive sites (M-DHS; UIA, UIIA) and three M-DHS (DIA, DII, DIII) were found within 2 kb upstream and 7 kb downstream regions, respectively. Two minor DHS (m-DHS; UIB, UIIB) in the upstream region and one m-DHS (DIB) downstream were shown to be associated with M-DHS. Thus, a total of five M-DHS and three m-DHS were mapped on the rat GH gene locus. Among these, five (UIIB, UIA, UIB, DIB, DIA) including two (UIA, DIA) M-DHS were specific for GH-producing cells. UIIA and DIII were M-DHS only in PRL-producing 235 cells while the major hypersensitivity of DII was detected in GH-producing cells and liver cells. Assessment of the enhancing activity of the DHS regions indicated novel enhancers in one upstream and two downstream regions that function well with the GH promoter in GC cells. These enhancers, each appearing different, coincided with m-DHS but not M-DHS in GC cells, and were not activated by Pit-1. Based on these observations, the following functions of five M-DHS and three m-DHS regions were defined: enhancer; locus control region (LCR); switch region serving for conversion from GH/PRL-producing cells to PRL-producing cells; and a region having a structural function in chromatin.

Animals↗
PubMed1995

Modulation by bovine angiogenin of tubular morphogenesis and expression of plasminogen activator in bovine endothelial cells.

Angiogenin is a potent angiogenic molecule in the chick chorioallantoic membrane assay and rabbit corneal assay. However, no angiogenic activity has been reported in in vitro system. In this study, we isolated and purified angiogenin from bovine milk, and the biological effect of the bovine angiogenin on bovine endothelial cells was examined in in vitro angiogenesis models. Bovine angiogenin significantly stimulated both cell migration and formation of tube-like structures in the collagen gel by bovine aortic endothelial cells. Angiogenin at 10-100 ng/ml stimulated about 2-fold higher the tube formation when basic fibroblast growth factor (bFGF) at 10 ng/ml stimulated about 3-fold over the control. Tubular morphogenesis stimulated by bFGF or angiogenin was almost completely blocked in the presence of aprotinin, an inhibitor of serine proteases. Angiogenin up-regulated both mRNA level and activity of urokinase type plasminogen activator, a key mediator of angiogenesis. Moreover, both bFGF and bovine angiogenin induced a marked increase of c-fos mRNA level at 30 min after stimulation. These novel effects of bovine angiogenin are to be discussed in relation to its structure specificity.

Allantois↗
PubMed1995

Methamphetamine plus scopolamine potentiates behavioral sensitization and conditioning.

The effects of repeated methamphetamine (4.0 mg/kg) plus scopolamine (0.5 mg/kg) treatment on behavioral sensitization and drug conditioning in rats were compared with the effects of repeated methamphetamine treatment. Behavioral sensitization induced by repeated methamphetamine plus scopolamine treatment was more vigorous than that induced by repeated methamphetamine treatment. Repeated methamphetamine plus scopolamine treatment produced sensitized responses, not only to methamphetamine plus scopolamine and methamphetamine but also, to a lesser extent, to scopolamine. Methamphetamine plus scopolamine-sensitized rats but not methamphetamine-sensitized rats exhibited conditioned responses to a low-frequency tone (300 Hz, 100 dB) associated with the drug state, suggesting that robust methamphetamine plus scopolamine-induced behavioral sensitization may lead to enhanced conditioning. It is plausible that robust behavioral sensitization might operate via a reciprocal balance between the dopaminergic and cholinergic systems in favor of dopaminergic dominance. Conditioning to the drug-associated tone may be mediated via a reciprocal balance between the two transmitter systems.

Acoustic Stimulation↗
PubMed1995

Effect of a nitric oxide synthase inhibitor, N-nitro-L-arginine methylester, on light-induced phase delay of circadian rhythm of wheel-running activity in golden hamsters.

Under constant darkness hamsters demonstrate free-running activity rhythms and light exposure during the early subjective night results in permanent phase delays of the activity rhythm. Recently, we reported that application of glutamate receptor agonists such as N-methyl-D-aspartate could reset the phase of the circadian rhythm of suprachiasmatic nucleus firing activity in vitro via nitric oxide production. In order to confirm this result by in vivo experiment, we examined the effect of nitric oxide synthesis inhibitor on the light-induced phase delay of circadian rhythms of wheel-running activity in hamsters. In vehicle-treated animals, light stimulation at circadian time 13.5 resulted in stable phase delays (1.3 +/- 0.63 h), whereas pre-treatment with 150 mg/kg of N-nitro-L-arginine methylester (L-NAME) significantly attenuated light-induced phase delays (0.72 +/- 0.18 h). L-NAME administration alone without light exposure, did not cause phase changes. The L-NAME-induced attenuating effect was reversed by co-administration of L-arginine (300 mg/kg). The present results suggest that nitric oxide production is involved in the light-induced phase delay of the hamster's circadian system.

Animals↗
PubMed1995

Long-lasting increase in protein kinase C activity in the hippocampus of amygdala-kindled rat.

Previous studies have demonstrated that membrane-associated protein kinase C (PKC) activities in the right and left hippocampus of rats kindled from the left hippocampus increased significantly at 4 weeks [9] and 4 months [22] after the last seizure compared with those in matched control rats. In this study, we investigated the effect of kindling from the left amygdala on PKC activities in the amygdala/pyriform cortex and hippocampus at long seizure-free intervals (4 and 16 weeks) from the last amygdala-kindled seizure. Membrane-associated PKC activity of the kindled group increased significantly only in the left hippocampus compared with the left side control (the left hippocampus of rats subjected to a sham operation) at 4 weeks (by 34%, P < 0.03) and 16 weeks (by 24%, P < 0.05) after the last seizure. There was no significant alteration in the membrane-associated PKC activity of the kindled group in the right hippocampus or amygdala/pyriform cortex in any seizure-free interval after the last amygdala seizure. Cytosolic PKC activity did not differ between the kindled and control groups in any brain region examined in any seizure-free interval. At 16 weeks after the last seizure, the PKC activity in the P1 fraction of the kindled group increased significantly only in the left hippocampus (by 49%, P < 0.005), but not in the right hippocampus. Neither PKC activity in the P2 fraction nor that in the cytosolic fraction was altered in the kindled group after this seizure-free interval.(ABSTRACT TRUNCATED AT 250 WORDS)

Amygdala↗
PubMed1995

The C/EBP site in the feline immunodeficiency virus (FIV) long terminal repeat (LTR) is necessary for its efficient replication and is also involved in the inhibition of FIV LTR-directed gene expression by pseudorabies virus ICP4.

We investigated effects of site-specific mutation of the putative C/EBP binding site in the feline immunodeficiency virus (FIV) long terminal repeat (LTR) on the basal promoter activity in Crandell feline kidney (CRFK) cells and on replication efficiency in CRFK cells and a T-lymphoblastoid cell line, MYA-1 cells. Mutation of the C/EBP site reduced the basal promoter activity in CRFK cells and prevented efficient FIV replication in both CRFK and MYA-1 cells. Gel-mobility-shift assay using nuclear extracts from CRFK and MYA-1 cells revealed that the nuclear factor(s) actually binds to the C/EBP site, but there was a clear difference in the binding patterns to the C/EBP site between CRFK and MYA-1 cell nuclear proteins. Furthermore, we demonstrated that the C/EBP site is necessary for inhibition of FIV LTR-directed gene expression by pseudorabies virus (PRV) ICP4. The C/EBP site is sufficient to confer inhibitory effect by PRV ICP4 on heterologous promoters. These data suggest that the C/EBP site in the FIV LTR is important for the positive regulation of FIV gene expression and replication and is also required for the negative regulation of FIV gene expression by PRV ICP4.

Animals↗
PubMed1995

Direct tumorigenic conversion of human gallbladder carcinoma cells by v-src but not by activated c-H-ras oncogene.

The roles of activated ras and src oncogene products in the acquisition of fully neoplastic phenotype by human gallbladder adenocarcinoma cells were investigated by co-transfecting non-tumorigenic HAG-I human gallbladder carcinoma cells with the pSV2neo plasmid and a plasmid carrying either activated c-H-ras or v-src oncogene. G418-resistant clones were isolated and assessed for the acquisition of anchorage-independent growth potential. Neither the 10 established clones transfected with pSV2neo alone nor the 17 clones transfected with activated c-H-ras, including 4 clones expressing the mutated p21H-ras protein, could form colonies in soft agar. By contrast, out of 10 clones transfected with v-src, 2 formed colonies in soft agar and produced tumors in athymic nude mice, the resulting progressive neoplasms being poorly differentiated adenocarcinomas. These tumorigenic clones were shown to have v-src DNA and mRNA levels with p60v-src protein, but there were no significant chromosomal alterations following tumorigenic conversion. Moreover, herbimycin A, a selective src-kinase inhibitor, markedly reduced clonogenic growth of these cells in soft agar rather than monolayer growth, suggesting that anchorage-independent growth of the v-src-transformed HAG-I cells might be driven directly by p60v-src kinase activity. Taken together, our data suggest that the fully neoplastic conversion of HAG-I cells depends on src-related tyrosine-kinase activity, but not solely on the function mediated by activated ras, thus providing evidence of an src-related signaling pathway for the acquisition of tumorigenic potential by human gallbladder adenocarcinoma cells.

Adenocarcinoma↗
PubMed1995

Expression of vascular endothelial growth factor and its possible relation with neovascularization in human brain tumors.

To examine which growth factors correlate with neovascularization in human brain tumors, the mRNA levels of transforming growth factor alpha, transforming growth factor beta, basic fibroblast growth factor, and vascular endothelial growth factor (VEGF) genes were determined by a Northern blot analysis in surgically obtained human gliomas and meningiomas. The vascular development was determined by counting the number of microvessels which were immunostained with von Willebrand factor. We normalized the growth factor mRNA levels versus the glyceraldehyde phosphate dehydrogenase mRNA level. In the 17 gliomas and 16 meningiomas examined, the mRNA of transforming growth factors alpha and beta, basic fibroblast growth factor, and VEGF were expressed at various levels. Among those 4 growth factors, the mRNA levels of VEGF, but not those of transforming growth factors alpha and beta and basic fibroblast growth factor, correlated significantly with vascularity in both gliomas (correlation coefficient r = 0.499; P < 0.05) and meningiomas (correlation coefficient r = 0.779; P < 0.001). These findings thus suggest that VEGF may be a positive factor in tumor angiogenesis in both human gliomas and meningiomas.

Brain Neoplasms↗
PubMed1995

The regulatory region and transcription factor required for the expression of rat and salmon pituitary hormone-encoding genes show cell-type and species specificity.

The promoter regions of the genes encoding the rat and chum salmon growth hormones (GH) and rat prolactin (PRL) were combined with a reporter gene and introduced into GH- and/or PRL-producing cells from rat. The rat GH and PRL promoters (pGH and pPRL, respectively) were most active in cells producing GH and PRL, respectively. The activity of the salmon pGH was much less than that of the rat pGH in rat GH-producing cells. The regulatory region required for cell-type-specific gene expression of pituitary hormones thus contains information, not only for cell-type specificity, but possibly for species specificity as well. A reporter plasmid containing the GH or somatolactin (SL) promoter and an effector plasmid having a gene encoding transcription factor Pit-1 (rat or salmon) were cotransfected into HeLa (human) or EPC (carp) cells. Rat and salmon Pit-1 were more active in HeLa and EPC cells, respectively, indicating that Pit-1 appears to interact species specifically with the transcription machinery.

Animals↗
PubMed1995

Structure, stereochemistry, and thermal isomerization of the male sex pheromone of the longhorn beetle Anaglyptus subfasciatus.

Male-released sex pheromone constituents of the longhorn beetle Anaglyptus subfasciatus (Coleoptera: Cerambycidae) are identified by GC-MS and GC-Fourier transform infrared as a 7:1 molar mixture of 3-hydroxy-2-hexanone and 3-hydroxy-2-octanone. These two compounds undergo thermal isomerization during GC analyses to give the corresponding 2-hydroxy-3-alkanones. Comparison of GC retention times of the natural products with those of synthesized enantiomerically pure compounds revealed that both semiochemicals have (R)-stereochemistry. These absolute configurations were confirmed by comparisons of the (R)-methoxy(trifluoromethyl)phenylacetic acid esters of insect-derived and synthetic samples.

Animals↗
PubMed1995

Expression of tissue-type plasminogen activator and its inhibitor couples with development of capillary network by human microvascular endothelial cells on Matrigel.

Human omental microvascular endothelial (HOME) cells seeded on Matrigel begin to migrate within 1 h, forming honeycomb-like structures and capillary-like networks within 18 h. Cross-sections of the capillary networks show them to be tube-like structures. Northern blot analysis showed that tissue-type plasminogen activator (t-PA) mRNA synthesis increased from the initial state at 0 h after seeding on Matrigel, reaching a steady state after 4 h. This elevated cellular t-PA mRNA level decreased markedly at 24 h. In contrast, the cellular plasminogen activator inhibitor-1 (PAI-1) mRNA level demonstrated biphasic curves during the 24 h after seeding on Matrigel: the PAI-1 mRNA level was increased eightfold initially at 4 h over that at 0 h, then declined, and again secondarily increased to greater than tenfold at 18 h. Cellular levels of both 72 kD type IV collagenase and tissue inhibitor of metalloproteinase (TIMP-2) mRNA were increased only a slightly within 2-4 h. These elevated mRNA levels were maintained for 18 h, while the TIMP-1 mRNA level increased up to 18 h, reaching around three times the level at 0 h. However, on collagen-coated dishes, cellular levels of t-PA, PAI-1, 72 kD type IV collagenase, TIMP-1, and TIMP-2 mRNA were not greatly changed during incubation for 24 h. On Matrigel, the cellular t-PA mRNA level at 18 h after seeding was greatly increased when treated with specific anti-transforming growth factor-beta (TGF-beta) antibody. In contrast, both PAI-1 and TIMP-1 mRNA levels at 18 h were reduced in the presence of anti-TGF-beta antibody. Development of the capillary network on Matrigel was inhibited in the presence of anti-t-PA antibody. Epidermal growth factor (EGF) enhanced t-PA gene expression and TGF-beta inhibited its expression in HOME cells cultured on collagen-coated dishes. On the other hand, TGF-beta enhanced cellular expression of the PAI-1 gene. The formation of a capillary network by HOME cells on Matrigel appears to be balanced by angiogenic EGF and anti-angiogenic TGF-beta through modulation of PA activity.

Capillaries↗
PubMed1995

Distribution of galanin-immunoreactive nerve fibers in the carotid labyrinth of the bullfrog, Rana catesbeiana: Comparison with substance P-immunoreactive fibers.

Immunoreactivity of galanin (GAL) was detected in the nerve fibers distributed within the intervascular stroma of the bullfrog carotid labyrinth. GAL-immunoreactive fibers are numerous, and some are close to the sinusoidal plexus. Most GAL fibers appear as thin processes with some varicosities. A combination of indirect double immunofluorescence labelling and image processing clearly demonstrated that the distribution pattern of GAL fibers is different from that of SP fibers. This indicates that GAL and SP do not coexist in the same nerve fibers. The role of GAL fibers may be different from that of previously reported neuropeptides (substance P, calcitonin gene-related peptide, vasoactive intestinal polypeptide, neuropeptide Y, and others) as a neuromodulator in controlling vascular tone of the labyrinth.

Animals↗
PubMed1995

Interaction between external Na+ and mexiletine on Na+ channel in guinea-pig ventricular myocytes.

To assess the modulation of Na+ channel block with local anaesthetics by the change of external Na+ concentration ([Na+]o), we examined the block by mexiletine at different [Na+]o using the whole-cell and the cell-attached configurations of the patch-clamp technique. Lowering [Na+]o increased the degree of use-dependent block of the whole-cell Na+ current. The external Na+ dependence of the Na+ current block was caused by the interaction of mexiletine with the activated Na+ channel, but not with the inactivated channel. In single-Na+ channel current recordings at a reduced [Na+]o of 70 mM, mexiletine shortened the mean open time of the channels (1.32 +/- 0.06 ms in the control vs. 0.86 +/- 0.12 ms with the drug, P < 0.05) without changes in the unitary current amplitude, whereas the drug did not affect mean open time at a [Na+]o of 140 mM. Moreover, the open time distributions during drug exposure at the reduced [Na+]o were better fitted to a double exponential than to a single exponential in four out of six experiments. These data suggest that mexiletine induces two conductive states: the native open state and a state representing the first step of open channel block. The transition from the former to the latter is dependent on [Na+]o, suggesting an antagonistic interaction of external Na+ with mexiletine.

Algorithms↗
PubMed1995

Usefulness of electron microscopy in the diagnosis of cardiac sarcoidosis.

A 49-year-old man with cardiac sarcoidosis is presented. He suffered from congestive heart failure, and left ventricular asynergy and reduced function was evident by echocardiogram and left ventriculogram. A light microscopic examination of the endomyocardial biopsy revealed nonspecific myocarditis without giant cells or noncaseating granulomas. Under an electron microscope, however, several epithelioid cells were found in the specimen. The serum level of lysozyme was elevated. The patient had a past history of sarcoidosis of the eyes and lungs 22 years previously. Cardiac diseases presenting epithelioid cells other than sarcoidosis were clinically ruled out. Thus, the diagnosis of cardiac sarcoidosis was made based on both clinical and ultrastructural findings, and corticosteroid therapy was initiated. In the second biopsy, performed 4 months later, a noncaseating granuloma was found. Generally, the incidence of histological diagnosis of cardiac sarcoidosis by light microscopy is relatively low in endomyocardial biopsy specimens. The present case suggests that the addition of an ultrastructural examination may improve the diagnostic usefulness of the endomyocardial biopsy in cardiac sarcoidosis, since electron microscopy can clearly identify the presence of even one epithelioid cell.

Biopsy, Needle↗
PubMed1995

Changes in free radical-metabolizing enzymes and lipid peroxides in the liver of Long-Evans with cinnamon-like coat color rats.

We report changes in free radical-metabolizing enzymes and the increased generation of lipid peroxides associated with extreme metal accumulation in the liver of the Long-Evans with cinnamon-like coat color (LEC) rat, a new mutant strain displaying hereditary hepatitis and subsequent hepatocellular carcinoma. The activity of free radical-metabolizing enzymes and lipid peroxides, and the concentration of metal in the liver were determined sequentially after birth. Mn-superoxide dismutase activity significantly increased immediately after the onset of hepatitis in LEC rats, whereas no remarkable change was observed in control rats. Cu, Zn-superoxide dismutase activity in LEC rats was similar to that in control rats. Glutathione reductase activity increased, while glutathione peroxidase activity was lower in LEC rats than in control rats throughout the observation periods. Lipid peroxides, estimated by thiobarbituric acid reaction, also increased 4- to 5-fold immediately after the onset of hepatitis in LEC rats. Copper concentration was 30- to 50-fold higher in the liver of LEC rats than in control rats, and the iron content also increased significantly before and after the onset of hepatitis. These findings suggested that an oxidant injury generated by toxic metals could be one of the factors responsible for hepatocellular damage in this unique hereditary hepatitis.

Analysis of Variance↗