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Biomedical subjects

S Murakami

Publications and source records attributed to S Murakami.

At least 577 records · Page 32Linked to original sources

c-myc gene in a murine plasmocytoma without visible chromosomal translocations moves to chromosome 12F1 with Pvt-1 and rearranges with IgH enhancer-S mu sequences.

The DCPC 21 plasmocytoma lacks any of the MPC-associated chromosomal translocations. However, the c-myc gene has been transposed to the IgH locus on chromosome 12 by an Ig switch-region-mediated recombination mechanism. DNA sequencing analysis, further, revealed that this recombination is consistent with an insertion of the IgH enhancer (E mu)-S mu sequences, 2341 bp in length, into the c-myc 5'-flanking region, resulting in 5': c-myc 5'-flanking-E mu-S mu-c-myc 5'-flanking-c-myc exon-1: 3' segment. In situ molecular hybridization of DCPC 21 metaphase chromosome spreads using a Pvt-1 probe demonstrated that Pvt-1 has also moved to the F1 sub-band region of chromosome 12 where the IgH genes are located. These results indicate that the c-myc gene has been inserted into the IgH locus together with the Pvt-1, regardless of whether plasmocytoma has cytogenetically identifiable translocations. The possible interaction between c-myc activation and Pvt-1 in the development of MPCs is discussed.

Animals↗

Hepatitis B virus X protein transactivates human interleukin-8 gene through acting on nuclear factor kB and CCAAT/enhancer-binding protein-like cis-elements.

Interleukin-8 (IL-8) is a newly described leukocyte chemotactic and activating cytokine that belongs to the novel family of inflammatory cytokines whose genes locate on human chromosome 4, q12-21 region. The production of IL-8 is usually not constitutive and can be induced rapidly and abundantly in different cell types by a variety of stimuli such as lipopolysaccharide, interleukin-1, tumor necrosis factor-alpha as well as a tumor promotor phorbol myristate acetate. We report here that in addition to these stimuli the IL-8 gene can also be induced by the protein X of the hepatitis B virus (HBV-X) as evidenced by the enhanced IL-8 mRNA expression and IL-8 production observed in HBV-X-transfected cells. Furthermore, using several deletion mutants of the 5'-flanking regulatory region of the human IL-8 gene linked to the chloramphenicol acetyl transferase gene as a reporter, we have established here that both nuclear factor kB and CCAAT/enhancer-binding protein-like cis-elements located at -94 to -71 base pairs of IL-8 gene are essential and sufficient for the induction of the IL-8 gene by HBV-X. The same elements have been identified recently by us to be interleukin-1-, tumor necrosis factor-alpha-, and phorbol myristate acetate-responsive elements on the IL-8 gene. This suggests the existence of a common pathway for these inflammatory cytokines and HBV-X to activate the IL-8 gene. These observations might be relevant to the pathogenesis of inflammation in viral hepatitis.

Base Sequence↗

Calcium oscillation associated with reduced protein kinase C activities in ras-transformed NIH3T3 cells.

We show here novel intracellular Ca2+ oscillation in v-K-ras-transformed NIH3T3 cells induced by mitogenic peptide hormones, bradykinin and bombesin, as well as fetal calf serum. Induction of the Ca2+ oscillation is strongly correlated with the malignant properties and inversely with PKC activities in vitro and in vivo. These results suggest that the mitogen-induced Ca2+ oscillation is negatively regulated by PKC, which modulates Ca2+ influx in v-K-ras-transformed NIH3T3 cells.

Animals↗

A large number of tRNA genes are symmetrically located in fission yeast centromeres.

We report here that the fission yeast centromere regions in the three chromosomes contain no less than 36 symmetrically arranged tRNA-coding sequences, and many of them are located within the inner inverted regions that are thought to be essential for the centromere function. There are 11 different species of tRNA-coding sequences, and four of them are identical to those previously known in this organism. This high-density distribution of tRNA genes in the centromere regions is surprising, as the fission yeast centromeres were thought to form transcriptionally inactive structures.

Base Sequence↗

Characterization of autoantibody-secreting B cells in mice undergoing stimulatory (chronic) graft-versus-host reactions. Identification of a CD44hi population that binds specifically to hyaluronate.

This study was undertaken to identify and isolate the pathophysiologically important B cell subpopulation which is activated to Ig secretion and autoantibody production in stimulatory (chronic) graft-vs-host (GVH) reactions. We recently demonstrated that IL-5 stimulation in vitro induces the appearance of a distinct CD44hi Ialow CD45Rlow B cell subpopulation that has aquired the ability to bind to hyaluronate (HA), one of the ligands for CD44, and that this B cell subpopulation is enriched in both proliferative and Ig-secretory responses. In the present report, CD44 expression was examined in B cells which were activated in the course of stimulatory GVH reactions. Compared with normal mice, B cells from mice undergoing stimulatory GVH reactions contained a novel CD44hi B cell subpopulation which exhibited enhanced binding to HA. The CD44hi HA-adherent B cell subpopulation from GVH mice spontaneously secreted large amounts of Ig, particularly IgG, including autoantibody specific for ssDNA. These findings demonstrate that CD44 expression distinguishes those B cells that are activated in vivo in the course of GVH to proliferate and differentiate into Ig-secreting cells. These CD44hi, HA-adherent, cells may play a prominent role in the hypergammaglobulinemia and immune complex glomerulonephritis that occur during chronic GVH reactions.

Animals↗

Cloning and expression in yeast of a cDNA clone encoding Aspergillus oryzae neutral protease II, a unique metalloprotease.

The neutral protease II (NpII) from Aspergillus oryzae is a zinc-containing metalloprotease with some unique properties. To elucidate its structure, we isolated a full-length cDNA clone for NpII. Sequence analysis reveals that NpII has a prepro region consisting of 175 amino acids preceding the mature region, which consists of 177 amino acids. As compared with other microbial metalloproteases, NpII is found to be unique in that it shares only a limited homology with them around two zinc ligand His residues and that the positions of the other zinc ligand (Glu) and the active site (His) cannot be established by homology. When a plasmid designed to express the prepro NpII cDNA was introduced into Saccharomyces cerevisiae and the transformant was cultured in YPD medium (2% glucose, 2% polypeptone, 1% yeast extract), it secreted a proNpII. However, in a culture of the same medium containing 0.2 mM ZnCl2, it secreted a mature NpII with a specific activity and N-terminus identical to those of native NpII. This observation suggests that either an autoproteolytic activity or a yeast protease effected the processing.

Amino Acid Sequence↗

Evidence for binding of at least two factors, including T-rich strand-binding factor(s) to the single-stranded ARS1 sequence in Saccharomyces cerevisiae.

To study the mechanism of initiation of eukaryotic chromosomal replication, we examined protein factors interacting with the ARS1 region located near the centromere of chromosome IV in Saccharomyces cerevisiae. Using the gel shift assay, we found protein factor(s) which specifically bound to the T-rich strand of the region containing the core consensus and its flanking sequences in ARS1, but not to the opposite strand. We designated this factor ATS (ARS1, T-rich strand-binding factor(s]. Similar specific complexes were also detected with oligonucleotide probes specific for the H4 or C2G1 ARS. As we have previously identified another binding factor, we conclude that at least two factors bind to the single-stranded ARS1 sequence.

Base Sequence↗

Structure of the fission yeast centromere cen3: direct analysis of the reiterated inverted region.

We determined the structure of the Schizosaccharomyces pombe centromere cen3 using direct genomic mapping and cosmid walking. The repetitive region of cen3 is approximately 110 kb, much longer than that of the previously determined cen1 and cen2 regions. The approximately 30 kb long left and approximately 60 kb right repetitive sequences are arranged with an inverted symmetry and flank the 15 approximately 20 kb central domain. The repeat motifs in cen3, although they consist of the common centromeric repeat elements, are slightly different from those in cen1 and cen2. The cen3 repeat motifs appear to be reiterated four times in the left and nine times in the right side repetitive regions. We found that the central domain consists of the common approximately 5 kb core sequence associated with the pair of innermost inverted sequences, most of which are reiterated only twice in the genome. Although their sizes differ significantly, the general features of cen1, cen2 and cen3 are similar, and a prototype, consensus structure for the fission yeast centromere may be deduced.

Base Sequence↗

Effect of 2-guanidinoethanol on levels of monoamines and their metabolites in the rat brain.

The contents of monoamines and their metabolites in rat brains 3 hours after the intracerebroventricular injection of 6 mumol of 2-guanidino-ethanol (GEt) were measured by HPLC. GEt which is a configurational analogue of 4-aminobutanoic acid (GABA) induced severe running fits and tonic-clonic convulsions as well as epileptic discharges. In GEt-administered rats, dopamine (DA) decreased in the cortex, hippocampus and hypothalamus. 3,4-Dihydroxyphenylacetic acid (DOPAC) increased to about the same level in all brain regions, therefore the distribution of DOPAC appeared to be homogeneous in the brain. The homovanillic acid levels also increased in the striatum and hippocampus. No significant change in the norepinephrine contents was observed in any region. The turnover ratio of DA increased significantly except in the striatum. Serotonin levels increased in the hypothalamus and midbrain by GEt administration, though 5-hydroxyindoleacetic acid levels showed no change in any of the brain regions. These data suggest that the activity of dopaminergic and serotonergic neurons are increased by GEt.

3,4-Dihydroxyphenylacetic Acid↗

False hyperchloremia in bromism.

Plasma chloride concentration measured by an ion-specific electrode can be interfered by other ions. The authors experienced a case of phantom limb pain with a marked hyperchloremia (251 mEq. l(-1)) which was measured by the ion-specific electrode method. The patient was diagnosed as bromide intoxication due to chronic ingestion of analgesic tablets which contain bromvalerylurea. A toxic level of plasma bromide concentration supported the diagnosis. Elevated plasma chloride and bromide concentrations were normalized in three weeks after discontinuation of the analgesic intake. Laboratory study revealed that fluoride ion did not affect chloride concentration measured by an ion-specific electrode. Bromide and iodide ions, however, interfered with the electrode and produced a large overestimation of chloride concentration. Hyperchloremia should be interpreted carefully when chloride was measured by an ion-specific electrode method.

Journal Article↗

Corrosion properties of soldered joints. Part II: Corrosion pattern of dental solder and dental nickel-chromium alloy.

Six nickel-chromium (Ni-Cr) alloys joined with two solders were evaluated for degree of corrosion after they were immersed in corrosive solution for 100 days. The corroded surface of the Ni-Cr alloy and the cross section of the soldered joint were observed and the solution of nickel and copper (Cu) ions in each combination was measured. No corrosion was observed in the high-resistant Ni-Cr alloys with high Cr content. Pitting corrosion was seen on low-resistant Ni-Cr alloys, which had less Cr content. In the cross section of the joint, the joint of silver solder became porous by corrosion and the corrosion advanced especially along the interface. Solution of Ni ions was detected in all of the combinations of Ni-Cr alloys and solders with exception of the combination of high-resistant Ni-Cr alloys and gold solder. On the other hand, solution of Cu ions was detected in all of the combinations with silver solder.

Chromium↗

Converging eye movements evoked by microstimulation of the fastigial nucleus of macaque monkeys.

Stimulation of the dorsal part of the fastigial nucleus in macaques is known to evoke ipsilateral saccades, while those of the ventral part produce contralateral saccades. It was found that stimulation of the transitional zone moved the visual axis (eyes) to converge at an area (focus) in the oculomotor range, regardless of the initial eye position. These saccades were designated as 'converging saccades'. Converging saccades were directed to the focus, but the eyes did not attain the focus in one motion. Only the repetition of stimuli brought the eyes near the focus. When stimulation is applied to the transitional zone at progressively more ventral sites, the focus gradually shifted, starting from the ipsilateral hemifield to the contralateral hemifield by taking various routes.

Animals↗

Effects of fastigial stimulation upon visually-directed saccades in macaque monkeys.

When the initial eye position was changed by stimulating the fastigial nucleus prior to visually-directed saccades, monkeys could not compensate for the stimulation-induced movement. The line of sight missed the target location by a distance and direction almost equal to the vector of the evoked saccade. When the stimulation was delivered 75-130 ms after the target presentation, saccades were triggered prematurely. Their initial movement reflected only the evoked saccade in some response and reflected the vector sum of the evoked and visually-directed saccades in the other. In contrast, when stimulus latencies were greater than 130 ms. saccades started toward the target location. The visuomotor processing for saccades seemed to be completed during this period, which is approximately half the latency of normal saccades. When the stimulation was applied while the eyes were already in motion, the trajectories of the saccades were strongly modified and terminated in deviated locations. These results indicate that cerebellar output impulses are projected downstream to saccade-programming circuits where visual information has already been converted into motor-command signals.

Animals↗

The ontogeny of luteinizing hormone-releasing hormone (LHRH) producing neurons in the chick embryo: possible evidence for migrating LHRH neurons from the olfactory epithelium expressing a highly polysialylated neural cell adhesion molecule.

The development of neurons expressing luteinizing hormone-releasing hormone (LHRH) has been studied immunohistochemically in the chick embryo from the 3.5 embryonic day (ED) to the day of hatching. At ED-3.5, LHRH-immunoreactive neurons were first detected in the medial epithelium of the olfactory pit, but their appearance in the brain was delayed to ED-4.5. On EDs-6-7, cords of the LHRH-immunoreactive cells extended across the nasal septum towards the ventromedial forebrain with the olfactory nerve. By double staining for LHRH and, a highly polysialylated form of neural cell adhesion molecule (NCAM-H), the LHRH-positive neurons in the olfactory-forebrain system were found strongly NCAM-H-positive. At ED-8, a marked decrease in the number of LHRH-positive cells in the olfactory epithelium and a concomitant increase in the LHRH-positive cells in the forebrain area were noted. From ED-11 to the day of hatching, the majority of LHRH-positive neurons tended to move into their usual adult position, whereas the LHRH-positive cells had almost disappeared in the olfactory epithelium. No LHRH-immunoreactive neurons were found strongly positive to NCAM-H. These results suggest that LHRH neurons originate from the olfactory placode, then as they develop they migrate across the nasal septum and enter the forebrain with the olfactory nerve. The close association of NCAM-H with the developing LHRH neurons raises the possibility that NCAM-H plays some role in guiding the migrating LHRH neurons from the olfactory epithelium to the forebrain.

Animals↗

Antidopaminergic effects of the stereoisomers of N-[(1-alkyl-2- pyrrolidinyl)methyl]-5-sulfamoylbenzamides and -2,3-dihydrobenzofuran-7-carboxamides.

The stereoisomers of some N-[(1-alkyl-2-pyrrolidinyl)methyl]-5- sulfaoylbenzamides (3-8) and -2,3-dihydrobenzofuran-7-carboxamides (9-18) were prepared to compare their dopamine D2 receptor binding affinities (in vitro) and inhibitory effects on apomorphine-induced hyperactivity (in vivo). In the 1-ethyl substituted compounds of the two series, the stereoisomers with S absolute configuration at the 2-position of the pyrrolidine moiety (S enantiomer 3 and 2S diastereomers 9 and 10) were more potent in both of the above activities than those with R absolute configuration (R enantiomer 4 and 2R diastereomers 11 and 12, respectively), whereas the R enantiomer (8) was more potent than the S enantiomer (7) in the 1-n-hexyl-substituted-benzamides and the 2R diastereomers (15, 16, and 18) were more potent than the 2S diastereomers (13, 14, and 17) in the 1-n-butyl- and 1-n-hexyl-2,3-dihydrobenzofuran-7-carboxamides. It was found that the stereospecificity of the compound activities altered from the S configuration to the R configuration as the 1-alkyl side chain became longer in the two series. How these stereoisomers meet the configurational requirements to interact with the dopamine D2 receptors is also discussed.

Animals↗

Inhibition of gastric H+, K(+)-ATPase and acid secretion by ellagic acid.

The effects of ellagic acid on gastric H+, K(+)-ATPase, acid secretion, and the occurrence of gastric ulcers were studied. Ellagic acid inhibited hog gastric H+, K(+)-ATPase activity with a 50% inhibition at 2.1 x 10(-6)M; kinetic studies showed that the inhibition of H+, K(+)-ATPase by ellagic acid is competitive with respect to ATP and is noncompetitive with respect to K+. The effect on gastric ulcers was investigated by using a stress ulcer model. Intraperitoneal administration of ellagic acid at above 5 mg/kg markedly reduced the occurrence of gastric lesion. Ellagic acid significantly reduced acid secretion at the same doses. These results suggest that ellagic acid has a marked inhibitory effect on acid secretion and the occurrence of stress-induced gastric lesions, and these effects may be attributed to the inhibition of H+, K(+)-ATPase activity.

Adenosine Triphosphatases↗