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Biomedical subjects

Y Ohshima

Publications and source records attributed to Y Ohshima.

At least 91 records · Page 5Linked to original sources
PubMed1996

Spliceosomal introns in conserved sequences of U1 and U5 small nuclear RNA genes in yeast Rhodotorula hasegawae.

U1, U2, U4, U5, and U6 small nuclear RNAs are essential for precursor mRNA splicing. We previously found one and four mRNA-type, or spliceosomal introns, in the U2 and U6 genes, respectively, of yeast Rhodotorula hasegawae (Erythrobasidium hasegawianum). We proposed that U2 and U6 RNAs form a catalytic core for precursor mRNA splicing and that the introns in those small nuclear RNA genes may have been acquired through reverse splicing of an intron from a precursor mRNA into a catalytic site in small nuclear RNAs. In the present study, we analyzed U1, U4, and U5 genes in R. hasegawae. One spliceosomal intron was found in the U1 region forming base-pairs with a 5' splice site of a precursor mRNA. The U5 gene has two spliceosomal introns in the region that interacts with 5' and 3' splice sites. In contrast, the gene for U4 RNA, which is released from the spliceosome prior to the first step of the splicing reaction, has no intron. These results lend a further support to the proposed relation between presence and position of an intron in an small nuclear RNA gene and the function of the encoded small nuclear RNA.

Base Sequence↗
PubMed1995

Prostaglandin E2 at priming of naive CD4+ T cells inhibits acquisition of ability to produce IFN-gamma and IL-2, but not IL-4 and IL-5.

We investigated the effect of prostaglandin E2 on the acquisition of cytokine-producing ability by naive CD4+ T cells in human cord blood. Naive CD4+ T cells were stimulated for 3 days with mouse monoclonal anti-CD3 Ab, then washed and expanded in IL-2-containing medium for 3 more days. These activated T cells produced IL-2, IL-4, IL-5, and IFN-gamma upon stimulation with PMA and ionomycin. PGE2 added at priming of naive T cells inhibited the production of IL-2 and IFN-gamma, but not of IL-4 and IL-5, in a dose-dependent manner. This change in the cytokine production profile induced by PGE2 was maintained in T cells restimulated with anti-CD3 in the absence of PGE2, expanded by IL-2, and stimulated with PMA and ionomycin. The mRNA expression of IFN-gamma and IL-2, but not that of IL-4, was also decreased in these cells. Forskolin and dibutyryl cAMP had a similar effect. PGE2 must exist at an early stage of T cell activation to inhibit priming for IL-2 and IFN-gamma production. PGE2 also showed this effect, even in the presence of exogenous IFN-gamma, at the primary stimulation. These results indicate that PGE2 inhibits the acquisition of the ability to produce IL-2 and IFN-gamma by acting directly on naive T cells. Our results suggest that PGE2 plays a role in facilitating the development of the Th2-type cytokine production profile.

Animals↗
PubMed1995

Isolation of active ribozymes from an RNA pool of random sequences using an anchored substrate RNA.

To analyze the structure-function relationship of a ribozyme, RNA molecules with cleavage activities were isolated from a pool of RNAs that have a region of 14 random nucleotides for catalysis by a novel in vitro selection method. Active ribozymes from the pool were selected using a substrate RNA anchored to an agarose bead. After 7 or 8 selection cycles, the pool of the selected molecules showed cleavage activity against the substrate RNA. Structures and catalytic activities of the individual selected RNA molecules were then analyzed by cDNA cloning, in vitro transcription and cleavage reactions. All the active ribozymes were found to catalyze the same sequence-specific cleavage reaction as does the hammerhead ribozyme and contain the conserved nucleotide sequences of the hammerhead ribozymes.

Base Sequence↗
PubMed1995

Neural regulation of thermotaxis in Caenorhabditis elegans.

Thermal stimulus is an important environmental factor influencing animal behaviour. However, the mechanisms underlying thermosensation and thermal adaptation are poorly understood. The nematode Caenorhabditis elegans can sense a range of environmental temperatures and migrate towards the cultivation temperature on a thermal gradient. This modifiable thermotactic response provides an ideal system for studying the cellular and molecular processes involved in thermosensation and thermal information storage. We have identified neurons critical for thermotaxis by killing individual cells in live animals. The results indicate that an amphid sensory neuron, AFD, is a major thermosensory neuron. Some of the genetically defined cryophilic and thermophilic mutant phenotypes were mimicked when amphid interneurons AIY and AIZ, respectively, were killed, indicating that AIY is responsible for thermophilic movement and AIZ for cryophilic movement. We propose a neural model in which regulation of the activities of the two interneurons in opposite directions, depending on the cultivation temperature, is essential for thermotaxis.

Animals↗
PubMed1995

A case of Werner's syndrome associated with systemic lupus erythematosus.

The case of a 40-year-old woman with Werner's syndrome associated with systemic lupus erythematosus (SLE) is reported. The patient exhibited short stature, slender extremities, thinned hair, high-pitched voice, cataracts, ulceration of the fingers, and mental retardation. Malar erythema, photosensitivity, and proteinuria had been noted since age 34. The serum contained high titers of antibodies to dsDNA, Sm, nRNP, and SS-A/Ro. The simultaneous presence of Werner's syndrome and SLE could be a coincidental occurrence of the two diseases, although it might be due to an abnormality in replication or degeneration of DNA leading to the development of both diseases.

Adult↗
PubMed1995

Serum levels of interleukin 4 and soluble CD23 in children with allergic disorders.

UNLABELLED: In order to clarify the clinical significance of serum interleukin 4 (IL-4) levels, we measured serum IL-4 concentrations in allergic and non-allergic children using a highly sensitive sandwich ELISA. The limit of detection of the assay was 0.15 pg/ml in serum samples. Serum IL-4 was detected in 96.3% (53/55) of non-allergic controls, in 92.9% (183/197) of allergic children, in 70% (7/10) of cord blood samples and in 86.7% (26/30) of neonates. The IL-4 levels in sera from non-allergic controls were relatively constant during the ages examined and all samples were under 1.5 pg/ml. In allergic children, the serum levels of IL-4 were significantly elevated, particularly at age 13-24 months. The serum levels of IL-4 did not differ in children with different clinical manifestations of allergy, such as bronchial asthma, and atopic dermatitis. The serum level of soluble CD23 (sCD23) showed an age-dependent change in allergic and non-allergic children and was significantly higher in allergic than in non-allergic infants aged 7 to 12 months, but not in other age groups. There was no significant correlation among serum levels of IL-4, sCD23 and IgE. CONCLUSION: It is suggested that the measurement of serum IL-4 and sCD23 is helpful in the examination of allergic patients in infancy and early childhood, but neither the serum level of IL-4 nor sCD23 directly reflects in vivo IgE production.

Asthma↗
PubMed1995

Physical interaction with monocytes rescues human mature CD4+ T-cell lines from anti-CD3-induced apoptosis.

Crosslinking of the TcR-CD3 complex with immobilized anti-CD3 antibodies without sufficient co-stimulation induced cell death in human mature CD4+ T-cell lines. In these T cells, DNA fragmentation and morphological characteristics of apoptosis were seen. The anti-CD3-induced apoptosis was inhibited by co-culture with monocytes. The rescue signal provided by monocytes does not need to be present simultaneously with signals mediated by anti-CD3. When T cells were precultured with monocytes for 24 h before anti-CD3 stimulation and then the monocytes were removed from the culture, anti-CD3-induced T-cell apoptosis was also inhibited. To determine whether the monocyte-derived rescue signals were transduced by soluble factors or by direct cell-to-cell interaction with monocytes, we precultured T cells with monocytes separated by a micropore membrane which prevented T cell-monocyte physical interaction but not the diffusion of secreted molecules. In this system, rescue signals could not reach the T cells. To further assess the importance of physical interaction, we precultured T cells with fixed monocytes. T cells could not be rescued from apoptosis under these experimental conditions, either. The results considered collectively suggest that sufficient physical interaction with viable monocytes is important for the rescue of anti-CD3-induced apoptosis of CD4+ T cells.

Antibodies, Monoclonal↗
PubMed1995

Drosophila MAP kinase kinase suppresses the vulvaless phenotype of lin-3, let-23 and lin-45 mutations in Caenorhabditis elegans.

The vulva of the nematode Caenorhabditis elegans develops from the three vulval precursor cells (VPCs) that are induced by a signal from the gonadal anchor cell. This signal is thought to be mediated by a receptor tyrosine kinase (RTK) in the VPCs to a downstream signal transduction pathway. A mitogen-activated protein kinase kinase (MAPKK) has been found to be one of the major components of an RTK pathway in other organisms. We expressed a wild type and an activated cDNA of Dsor1, a Drosophila MAPKK, in each of the three vulvaless mutants lin-3, let-23 and lin-45. The expression of an activated from of Dsor1 in each of the mutants effectively induced a normal, functional vulva, that is, suppressed the vulvaless phenotype. The wild type Dsor1 also suppressed albeit less effectively. These results suggest that a MAPKK is involved in the vulval induction of C. elegans.

Animals↗
PubMed1995

Role of LFA-1/ICAM-1-dependent cell adhesion in CD40-mediated inhibition of anti-IgM antibody-induced B-cell death.

Cross-linking of surface IgM by anti-IgM antibody caused activation-induced cell death of a surface IgM+, IgD+ human B lymphoma cell line, B104. The dying B104 cells did not show the morphology of apoptosis but did show that of necrosis. However, anti-IgM antibody caused apoptosis of another surface IgM+, IgD+ human B lymphoma cell line, DND-39. The influx of extracellular Ca2+ was necessary for the cell deaths of B104 and DND-39 caused by anti-IgM antibody. Their cell deaths were inhibited by cyclosporine. The anti-IgM antibody-induced cell death of DND-39, but not that of B104, was prevented by costimulation with anti-CD40 antibody. In human peripheral blood B-cells, anti-IgM antibody inhibited cell cycle transition induced by Staphylococcus aureus Cowan I at the G2/M interphase without inhibition of DNA synthesis. In this system, too, anti-CD40 antibody canceled the inhibitory signal transduced through surface IgM and increased the number of M phase cells. Blocking antibodies against the leukocyte function-associated antigen-I/intercellular adhesion molecule-1 system decreased the rescue effect of anti-CD40 antibody in both DND-39 cells and peripheral B-cells, which shows that leukocyte function-associated antigen-1/intercellular adhesion molecule-1-dependent cell adhesion plays an important role in the CD40-mediated inhibition of surface IgM-mediated negative signals.

Animals↗
PubMed1995

Mosaic analysis of the let-23 gene function in vulval induction of Caenorhabditis elegans.

The let-23 receptor tyrosine kinase gene is required for vulval induction and larval survival in the nematode Caenorhabditis elegans. We carried out genetic mosaic analyses of the let-23 gene function by using the cloned let-23 and ncl-1 genes. The wild-type let-23 gene was required in a vulval precursor cell to adopt the 1 degree vulval fate in animals carrying a let-23 vulvaless or lethal chromosomal mutation. In almost all the animals, vulval precursor cells adjacent to a 1 degree fate cell were induced to the 2 degrees vulval fate regardless of the let-23 genotypes. These findings indicate that the vulval induction signal from an anchor cell induces a vulval precursor cell to adopt the 1 degree fate through LET-23, and then a 1 degree fate cell induces adjacent cells to adopt the 2 degrees fate, for which LET-23 is not required. Foci of lethality of the let-23 (mn23) mutation were found in ABal and ABplp lineages.

Animals↗
PubMed1995

IgM-mediated B cell apoptosis.

Cross-linking of surface immunoglobulin M (sIgM) on normal mature B cells induces different signaling consequences, including DNA synthesis (positive signaling) and cell cycle arrest and/or death by apoptosis (negative signaling). Presumably, the difference depends on the intensity of sIgM cross-linking: relatively weak cross-linking induces DNA synthesis, moderate cross-linking induces DNA synthesis with cell cycle arrest at the G2/M interphase, and intense cross-linking induces apoptosis. In vivo experiments with transgenic mice have shown that relatively weak cross-linking of sIgM by soluble antigens induces anergy in autoreactive B cells, whereas intense sIgM cross-linking by membrane-bound forms of antigens induces deletion of them. However, it is still unknown whether the different intensities of sIgM cross-linking generate qualitatively different signals responsible for DNA synthesis or cell death or whether they generate qualitatively the same but quantitatively different signals, and the quantitative difference is responsible for the induction of positive or negative signaling. The sIgM-mediated negative signaling presumably plays an important role in the induction and maintenance of B cell tolerance, and sIgD and sIgG also possess the machinery necessary for negative signaling. Negative signaling through sIgM is dependent on tyrosine kinase(s) and Ca2+ influx and is sensitive to cyclosporin A in certain types of B cells but not in all B cells. It has been suggested that there are different intracellular signaling pathways that transduce negative signaling via sIgM, and that activation-induced B cell death by sIgM cross-linking does not necessarily show DNA fragmentation and the morphology of apoptosis. On the other hand, sIgM-mediated B cell death may be inhibited in the presence of appropriate co-stimulators such as IL-4, alpha-, and beta-interferons and CD40-mediated signaling. The CD40-mediated signaling effectively inhibits sIgM-mediated B cell apoptosis in many but not all experimental systems. Although homotypic cell adhesion through the LFA-1/ICAM-1 dependent pathway was shown to be involved in certain types of CD40-mediated inhibition of sIgM-mediated negative signaling, it is still not known how the cytokines and CD40-mediated signaling inhibit sIgM-mediated B cell death. The molecular mechanisms responsible for sIgM-mediated negative signaling and for the inhibitory signaling against sIgM-mediated negative signaling need further elucidation.

Animals↗
PubMed1995

[Inhibition of pulmonary vasoconstriction by inhaled nitric oxide].

We examined effects of inhaled NO gas on pulmonary vasoconstriction induced with hypoxic gas or PGF2 alpha utilizing the isolated perfused rabbit lung. While NO 180 ppm did not change the basal pulmonary artery pressure, the pressure response to hypoxia or PGF2 alpha was completely inhibited by pretreatment with NO inhalation. Inhaled NO during maximal pressor response also depressed the elevated pressure to the basal level. These findings suggest that NO inhaled to the alveoli diffuses directly into the pulmonary vascular smooth muscle and then causes nonspecific vasodilation, and its effect is more potent than that of constitutive type of EDRF/NO.

Administration, Inhalation↗
PubMed1995

[Osteoconduction in porous hydroxyapatite ceramics grafted into the defect of the lamina in experimental expansive open-door laminoplasty in the spinal canal].

We have investigated the osteoconduction in porous hydroxyapatite ceramics grafted into the defect of the lamina of dogs in expansive open-door laminoplasty. Porous Hydroxyapatite block (HAB) was sintered at 900 degrees C with size of 4 x 7 x 19 mm, a mean pore diameter of 90 microns and a void volume of 70%. The operation was performed in 29 dogs. The animals were sacrificed at three or at six months after the operation (the three-month group and the six-month group). After the grafted HABs with adjacent vertebrae were removed, the stability of the grafted HABs was manually evaluated. The HAB was firmly fixed to the laminae in 14 of 20 cases. It seemed that the stability of the HAB was poor in those cases in which the HAB had not been tightly sutured. To investigate osteoconduction, serial sections of the laminar bones including the HAB were prepared without decalcification and were examined histologically. New bone formation in the pores of the HAB was found in all cases. However, the osteoconduction rate was higher in the six-month group than in the three-month group (p < 0.01). And the osteoconduction rate was higher in those cases in which the HAB was firmly fixed to the laminae than in those with a loosely fixed HAB. Bone ingrowth in the HAB was frequently observed in the zone adjacent to the bone, more so than in other zones. Osteoconduction appeared to occur at the contact surface and to extend towards the center of the HAB.

Animals↗
PubMed1994

Caenorhabditis elegans unc-51 gene required for axonal elongation encodes a novel serine/threonine kinase.

Mutations in the unc-51 gene of the nematode Caenorhabditis elegans result in various abnormalities in axonal elongation and axonal structures. We cloned the unc-51 gene by tagging with the transposon Tc1. The wild-type unc-51 gene, which rescued the mutant phenotypes, encodes a novel serine/threonine kinase of 856 amino acids. Mutation sites were identified in the unc-51 gene of six mutants. A Lys-->Met mutation created in vitro in the kinase domain led to the loss of rescuing activity and was dominant negative, indicating that the kinase domain of Unc-51 is essential for the function. Expression of an unc-51/lacZ fusion gene was observed in many neurons at all stages. We propose that protein phosphorylation by the unc-51 product is important for axonal elongation and possibly for axonal guidance.

Alleles↗
PubMed1994

High-performance liquid chromatographic determination of the new quinolone antibacterial agent DU-6859a in human serum and urine using solid-phase extraction with photolysis-fluorescence detection.

A sensitive and specific HPLC method for the determination of DU-6859a (I), a fluoroquinolone antibacterial agent, in human serum and urine was developed. Compound I and the internal standard extracted from serum and urine by means of a Bond Elut C8 LRC cartridge showed recoveries of 96%. The extracts were chromatographed on a reversed-phase column with photolysis-fluorescence detection. This unique detection method was 42.5 times more sensitive than intrinsic fluorescence detection, the limits of detection being in 3.43 ng/ml for serum and 4.35 ng/ml for urine. In addition, I was stable in serum and urine for at least 1 month at -20 degrees C. The proposed method was sensitive and selective enough to apply to pharmacokinetic studies of I in humans after a single oral dose of 100 mg.

Anti-Infective Agents↗