Biomedical subjects

K Hashimoto

Publications and source records attributed to K Hashimoto.

At least 145 records · Page 8Linked to original sources

PubMed2000

Isolation of novel mouse genes differentially expressed in brain using cDNA microarray.

The major issue in the post-genome sequencing era is determination of gene expression patterns in variety of biological systems. A microarray system is a powerful technology for analyzing the expression profile of thousands of genes at one experiment. In this study, we identified highly expressed genes in mouse brain using the cDNA microarray carrying 2304 cDNAs derived from oligo-capped mouse cDNA library. Nine genes were highly expressed in adult mouse brain compared to kidney, liver, and skeletal muscle. Tissue distribution analysis by reverse transcription-coupled polymerase chain reaction (RT-PCR) revealed that consistent with the microarray data, all of the selected 9 genes were predominantly expressed in the brain. A database search showed that 5 of the 9 genes, MBP, SC1, HiAT3, S100 protein-beta, and SNAP25, were previously known to be expressed at high level in the brain and in the nervous system. One gene was highly sequenced similar to rat S-Rex-s/human NSP-C, suggesting that the gene is a mouse homologue. The remaining three genes did not match to known genes in the GenBank/EMBL database, indicating that these are novel genes highly expressed in the brain. Taken together, our cDNA microarray system can be an excellent tool for identifying differentially expressed genes in mouse brain.

Animals↗

PubMed2000

3,4-methylenedioxymethamphetamine (MDMA, ecstasy)-induced egr-1 mRNA in rat brain: pharmacological manipulation.

Using in situ hybridization and immunohistochemical techniques, we examined the expression pattern of egr-1 mRNA and Egr-1 protein in several brain regions following administration of 3, 4-methylenedioxymethamphetamine (MDMA). Furthermore, we also studied the role of N-methyl-D-aspartate (NMDA) receptor, dopamine D(1) receptor, 5-hydroxytryptamine (5-HT) transporter or 5-HT(2A) receptor in the induction of egr-1 mRNA by MDMA. Basal constitutive levels of egr-1 mRNA were detected in control rat brains. A single administration of MDMA (10 mg/kg) caused marked induction of egr-1 mRNA in the prefrontal cortex, striatum and hippocampal dentate gyrus. However, no changes in the egr-1 mRNA levels were detected in the CA1 region of hippocampus and occipital cortex after administration of MDMA (10 mg/kg). Furthermore, the expression of egr-1 mRNA in the prefrontal cortex, striatum and hippocampal dentate gyrus after administration of MDMA (10 mg/kg) was blocked significantly by pretreatment with NMDA receptor antagonist (5R, 10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,b]-cyclohepten-5, 10-imine ((+)-MK801; 1 mg/kg), dopamine D(1) receptor antagonist SCH 23390 (1 mg/kg) or 5-HT uptake inhibitor paroxetine (5 mg/kg), but not by 5-HT(2A) receptor antagonist SR46349B (5 mg/kg). However, high basal levels of Egr-1 immunoreactivity in the rat brain were not altered by administration of MDMA (10 mg/kg). These results suggest that MDMA alters the expression of egr-1 mRNA in several regions of rat brain, and that the expression of egr-1 mRNA by MDMA in the prefrontal cortex, striatum and hippocampal dentate gyrus appears to be mediated, at least in part, by NMDA receptor, dopamine D(1) receptor and 5-HT transporter.

Animals↗

PubMed2000

Long-term treatment with haloperidol decreases the mRNA levels of complexin I, but not complexin II, in rat prefrontal cortex, nucleus accumbens and ventral tegmental area.

The effect of long-term treatment with haloperidol on gene expression of the presynaptic protein complexins was investigated in the discrete brain regions of rats, using reverse transcription-polymerase chain reaction. Four-week-treatment with haloperidol decanoate (25 mg eq/kg) produced a significant decrease in the mRNA levels of complexin I in the medial prefrontal cortex, nucleus accumbens and ventral tegmental area, but not in the striatum and substantia nigra. No significant changes in complexin II mRNA levels were observed in any brain region examined here. The reduced expression of complexin I may be associated with the haloperidol-induced depolarization block of mesocorticolimbic dopamine neurons.

Adaptor Proteins, Vesicular Transport↗

PubMed2000

Cytokines prevent dexamethasone-induced apoptosis via the activation of mitogen-activated protein kinase and phosphatidylinositol 3-kinase pathways in a new multiple myeloma cell line.

A new human myeloma cell line, OPM-6, was established from the peripheral blood of a patient with advanced IgG-kappa plasma cell leukemia. Cytogenetic and phenotypic analysis confirmed that the cells were derived from the patient's leukemic cells. Insulin-like growth factor-1 (IGF-1) acts as an autocrine growth factor in these cells. In addition, OPM-6 cells were particularly sensitive to dexamethasone (DEX), when endogenous IGF-1 was blocked. Under these conditions, >95% of the DEX-treated cells died within 36 h. Therefore, OPM-6 represents a potentially powerful tool for the analysis of the molecular mechanisms of DEX-induced apoptosis, because it is possible to easily analyze the direct effects of DEX using this system. Using this culture system of OPM-6, we demonstrated that the treatment with DEX plus a monoclonal antibody to the human IGF-1 receptor (alphaIGF-1R) leads to the down-regulation of the gene expression of Bcl-xL, an antiapoptotic gene, and the activation of CPP32 during this apoptotic process. IFN-alpha as well as IL-6 prevented DEX plus alphaIGF-1R-induced apoptosis, and this prevention was blocked by the mitogen-activated protein kinase kinase inhibitor, PD098059, or the phosphatidylinositol 3-kinase inhibitor, wortmannin. Therefore, both IL-6 and IFN-alpha blocked DEX plus alphaIGF-1R-induced apoptosis through activation of the mitogen-activated protein kinase and phosphatidylinositol 3-kinase pathways.

Aged↗

PubMed2000

QT-prolonging class I drug, disopyramide, does not aggravate but suppresses adrenaline-induced arrhythmias. Comparison with cibenzoline and pilsicainide.

We investigated the effects of class I antiarrhythmic drugs on corrected QT (QTc) interval and adrenaline-induced arrhythmias in halothane-anaesthetized, closed-chest dogs. For this purpose, we plotted a dose-response curve for adrenaline by calculating the arrhythmic ratio, which is the number of ventricular ectopic beats induced by adrenaline divided by the total heart rate, and observed the changes in the arrhythmic ratio-adrenaline dose relation before and after administration of class I drugs. Disopyramide and cibenzoline decreased the arrhythmic ratio induced by adrenaline. Disopyramide prolonged the QTc interval by 20% (P<0.01), but cibenzoline did not. Pilsicainide prolonged the QTc interval (12%), but this drug did not change the arrhythmic ratio. These results indicate that in contrast to the class III drugs which we have reported earlier, i.e. 1, 3-dimethyl-6-2-[N-(2-hydroxyethyl)-3-(4-nitrophenyl)-propylamino]eth ylamino-2,4 (1H,3H)-pyrimidinedione hydrochloride (MS-551), 1-(2-amino-4-methanesulfonamidophenoxy)2-[N-(3, 4-dimethoxyphenethyl)-N-methylamino]ethane hydrochloride (KCB-328) and E-1-[(5-(4-chlorophenyl)-2-furanyl)methylene]amino-3-[4-(4-methyl-1 -piperazinyl)butyl]-2,4-imidazolidinedione dihydrochloride (azimilide), class I drugs do not aggravate adrenaline-induced arrhythmias even though some drugs prolong the QTc interval.

Animals↗

PubMed2000

Photocatalytic sensor for chemical oxygen demand determination based on oxygen electrode.

The construction and performance evaluation of a novel Chemical Oxygen Demand (COD) sensor is described. The sensor measures, using an oxygen electrode, a decrease of dissolved oxygen of a given sample resulting from photocatalytic oxidation of the organic compounds therein. As the photocatalyst, titanium dioxide (TiO2) fine particles adsorbed on a translucent poly(tetrafluoroethylene) (PTFE) membrane was used. The oxygen electrode with the membrane attached on its tip was used as the sensor probe. The operation characteristics of the sensor are demonstrated using an artificial wastewater and real water samples from lakes in Japan. This method is considered to be reliable, in that the observed parameter is close to the theoretical definition of chemical oxygen demand (COD), the amount of oxygen consumed for oxidation of organic compounds.

Biosensing Techniques↗

PubMed2000

Modulation of type I interleukin-1 receptor messenger RNA followed by one and repeated endotoxin treatment in the mouse.

The expression of transcripts for type 1 interleukin-1 receptors (IL-1R1) were investigated utilizing reverse transcription-polymerase chain reaction (RT-PCR) techniques in male C57BL/6 mice. First, we measured IL-1R1 mRNA 1, 2, 6, and 12 h after a single injection of endotoxin lipopolysaccharide (LPS; 30 microg/mouse). IL-1R1 mRNA was measured in relevant tissues utilizing semiquantitative RT-PCR. IL-1R1 mRNA levels were significantly increased 2 and 6 h after one LPS injection group in comparison with the saline injection group in the hippocampus and testis. Next, we investigated the effect of repeated injections of LPS on IL-1R1 expression. LPS or saline were injected at 24-h intervals for 4 consecutive days. The mice were divided into four groups: (1) LPS injections for 4 days and LPS injection on day 5 (LPS-LPS); (2) LPS injections for 4 days and saline injection on day 5 (LPS-saline); (3) saline injections for 4 days and LPS injection on day 5 (saline-LPS); (4) saline injections for 4 days and saline injection on day 5 (saline-saline). The mice were sacrificed by decapitation 2 h after the last injection. In the hippocampus and pituitary gland, significant increases of IL-1R1 mRNA levels were observed in the saline-LPS group, however, the increases were attenuated in LPS-LPS group. IL-1R1 mRNA levels in the hypothalamus were unchanged in all four groups. In the adrenal gland and testis, IL-1R1 mRNA levels were significantly increased in the LPS-LPS group as well as saline-LPS group. These data demonstrate differential regulation of IL-1R1 mRNA by one and repeated endotoxin treatment and suggest the importance of the cytokines in mediating the effects of infectious challenge on brain, endocrine and immune function during endotoxemia.

Adrenal Glands↗

PubMed2000

Human centromere protein A (CENP-A) can replace histone H3 in nucleosome reconstitution in vitro.

Centromere protein A (CENP-A) is a variant of histone H3 with more than 60% sequence identity at the C-terminal histone fold domain. CENP-A specifically locates to active centromeres of animal chromosomes and therefore is believed to be a component of the specialized centromeric nucleosomes on which the kinetochores are assembled. Here we report that CENP-A, highly purified from HeLa cells, can indeed replace histone H3 in a nucleosome reconstitution system mediated by nucleosome assembly protein-1 (NAP-1). The structure of the nucleosomes reconstituted with recombinant CENP-A, histones H2A, H2B, and H4, and closed circular DNAs had the following properties. By atomic force microscopy, "beads on a string" images were obtained that were similar to those obtained with nucleosomes reconstituted with four standard histones. DNA ladders with repeats of approximately 10 bp were produced by DNase I digestion, indicating that the DNA was wrapped round the protein complex. Mononucleosomes isolated by glycerol gradient sedimentation had a relative molecular mass of approximately 200 kDa and were composed of 120-150 bp of DNA and equimolar amounts of CENP-A, and histones H4, H2A, and H2B. Thus, we conclude that CENP-A forms an octameric complex with histones H4, H2A, and H2B in the presence of DNA.

Autoantigens↗

PubMed2000

Basophilic cytoplasmic inclusions in a case of sporadic juvenile amyotrophic lateral sclerosis.

A 24-year-old woman presented with progressive muscle atrophy and weakness of the right upper extremity. Subsequently her weakness rapidly extended to the left upper extremity, neck and lower extremities. Neurological examination disclosed involvement of the lower motor neuron system. She died 7 months after the onset. There was neuronal loss and reactive gliosis in the anterior horns of the spinal cord and much less frequently in the motor cortex. Basophilic cytoplasmic inclusions were observed in the thalamus and brain stem as well as the upper and lower motor neurons. Ultrastructurally, the inclusions lacked a limiting membrane and consisted of a meshwork of filamentous structures associated with granules. The inclusions failed to react with antibodies against phosphorylated neurofilament or cystatin C. Most of the inclusions show no reaction with anti-ubiquitin antibody, however, a few inclusions show granular reaction product deposits with this antibody. The inclusions were not immunostained with antibodies against TGN46 and MG-160, markers of the trans-Golgi network and the medial cisternae of the Golgi apparatus, respectively, suggesting that they were not derived from the Golgi apparatus which was fragmented.

Adult↗

PubMed2000

Effects of antipsychotic drugs on neurotoxicity, expression of fos-like protein and c-fos mRNA in the retrosplenial cortex after administration of dizocilpine.

In this study, we examined the effect of clozapine, olanzapine, risperidone and haloperidol on the neuropathology (i.e. neuronal vacuolization) and the expression of Fos-like protein and c-fos mRNA in the retrosplenial cortex of female Sprague-Dawley rats induced by the NMDA receptor antagonist dizocilpine. Pretreatment (15 min) with clozapine or olanzapine, but not risperidone or haloperidol, blocked the neuronal vacuolization produced by dizocilpine (0.5 mg/kg, s.c.) in the rat retrosplenial cortex in a dose-dependent manner. Furthermore, pretreatment (15 min) with clozapine or olanzapine, but not risperidone or haloperidol, significantly attenuated the expression of Fos-like protein in the retrosplenial cortex induced by dizocilpine (0.5 mg/kg, s.c.) in a dose-dependent manner. The marked expression of c-fos mRNA in the rat retrosplenial cortex induced by the administration of dizocilpine (0.5 mg/kg, s.c.) was significantly attenuated by pretreatment (15 min) with clozapine (10 mg/kg) or olanzapine (10 mg/kg), but not risperidone (10 mg/kg) or haloperidol (10 mg/kg). The present results suggest that pharmacologically relevant doses of clozapine or olanzapine, but not risperidone or haloperidol, block the neuropathological changes and the expression of Fos-like protein and c-fos mRNA in the rat retrosplenial cortex elicited by the administration of dizocilpine. It is possible that the blockade of dizocilpine-induced neuropathological changes by clozapine and olanzapine may be related to the unique antipsychotic actions of these drugs in schizophrenic patients, although this remains to be verified.

Animals↗

PubMed2000

Antiarrhythmic and cardiohemodynamic effects of a novel Ca(2+) channel blocker, AH-1058, assessed in canine arrhythmia models.

The antiarrhythmic profile and cardiohemodynamic effect of a novel Ca(2+) channel blocker, 4-(5H-Dibenzo[a, d]cyclohepten-5-ylidene)-1-[(E)-3-(3-methoxy-2-nitro)phenyl-2-p ropeny l]piperidine hydrochloride (AH-1058), were analyzed using the epinephrine-, digitalis- and two-stage coronary ligation-induced canine ventricular arrhythmia models. Intravenous administration of AH-1058 (100 microg/kg) effectively suppressed each of the ventricular arrhythmias accompanied by weak hypotensive effects. The results contrast well with those of a typical Ca(2+) channel blocker, verapamil, which suppresses only the epinephrine-induced ventricular arrhythmia with severe hypotension. These results indicate that AH-1058 may possess a more selective inhibitory action on Ca(2+) channels in the heart than on those in the vessels. Furthermore, the antiarrhythmic actions of AH-1058 were slower in onset and longer-lasting, than those in our previous studies using other antiarrhythmic drugs, including Na(+) and Ca(2+) channel blockers. The antiarrhythmic effects of AH-1058 did not correlate with its plasma concentrations when administered either intravenously or orally. These results suggest that AH-1058 can become a long-acting Ca(2+) channel blocker with unique antiarrhythmic properties, and that AH-1058 may be used in certain pathological processes, for which selective inhibition of the cardiac Ca(2+) channels is essential.

Animals↗

PubMed2000

mGluR1 in cerebellar Purkinje cells essential for long-term depression, synapse elimination, and motor coordination.

Targeted deletion of metabotropic glutamate receptor-subtype 1 (mGluR1) gene can cause defects in development and function in the cerebellum. We introduced the mGluR1alpha transgene into mGluR1-null mutant [mGluR1 (-/-)] mice with a Purkinje cell (PC)-specific promoter. mGluR1-rescue mice showed normal cerebellar long-term depression and regression of multiple climbing fiber innervation, events significantly impaired in mGluR1 (-/-) mice. The impaired motor coordination was rescued by this transgene, in a dose-dependent manner. We propose that mGluR1 in PCs is a key molecule for normal synapse formation, synaptic plasticity, and motor control in the cerebellum.

Animals↗

PubMed2000

Isolation and characterization of a novel zinc finger gene, ZNF219, and mapping to the human chromosome 14q11 region.

We have isolated and characterized a novel zinc finger gene by screening a human testis cDNA library. The isolated cDNA, termed ZNF219, contains an open reading frame of 2169 nucleotides encoding 723 amino acids. Sequence analysis revealed 9 sets of Kruppel-related zinc finger structures and proline-rich regions in several parts. ZNF219 exhibited ubiquitous expression in all fetal and adult tissues examined. The transcript size was 5.5 kb in adult tissues, while the main transcript size in the embryo stage was 3.5 kb. The transcript size is developmentally regulated. When the plasmid cloned with green fluorescent protein (GFP)-tagged ZNF219 was expressed in HeLa and COS7 cells, strong fluorescence intensities were observed only in the nucleus of both cells by fluorescence confocal microscopy. These data suggest that ZNF219 may be related to the regulation of transcription and developmental regulation. Genomic structure analysis mapped ZNF219 to chromosome 14q11 between markers D14S72 and D14S990, because a sequence tagged site mapped to the locus was found in the intron region of the ZNF219 gene.

Adult↗

PubMed2000

Primitive synteny of vertebrate major histocompatibility complex class I and class II genes.

Major histocompatibility complex (MHC) class I and class II molecules bind to and display peptidic antigens acquired from pathogens that are recognized by lymphocytes coordinating and executing adaptive immune responses. The two classes of MHC proteins have nearly identical tertiary structures and were derived from a common ancestor that probably existed not long before the emergence of the cartilaginous fish. Class I and class II genes are genetically linked in tetrapods but are not syntenic in teleost fish, a phylogenetic taxon derived from the oldest vertebrate ancestor examined to date. Cartilaginous fish (sharks, skates, and rays) are in the oldest taxon of extant jawed vertebrates; we have carried out segregation analyses in two families of nurse sharks and one family of the banded houndshark that revealed a close linkage of class IIalpha and beta genes both with each other and with the classical class I (class Ia) gene. These results strongly suggest that the primordial duplication giving rise to classical class I and class II occurred in cis, and the close linkage between these two classes of genes has been maintained for at least 460 million years in representatives of most vertebrate taxa.

Amino Acid Sequence↗

PubMed2000

Asymmetric synthesis of axially chiral biaryls via desymmetrization of 2,2',6,6'-tetrahydroxybiphenyl using 1,4-Di-O-benzyl-L-threitol as a chiral template

Sequential etherification of 2,2',6,6'-tetrahydroxybiphenyl (1) with 1,4-di-O-benzyl-L-threitol under Mitsunobu conditions gives desymmetrized biphenyldiol 9 of S-axial chirality exclusively. Cyclization of 9 with 1,omega-dibromoalkanes followed by removal of the chiral auxiliary yields (S)-2,2'-biphenyldiols 14 with alkylenedioxy bridges at the 6 and 6' positions. (S)-6,6'-Dialkyl- and -diphenyldiols 20 are obtained in an efficient manner via Pd(0)-catalyzed cross-coupling of bis(triflate) derivative 17 with organozinc reagents. Bis(triflate) 17 also serves as an intermediate for asymmetric synthesis of axially chiral biphenyldicarboxylic acid 23, terphenylcarboxylic acid 28, lactone 26, and lactam 30.

Journal Article↗

PubMed2000

An IP3-assisted form of Ca2+-induced Ca2+ release in neocortical neurons.

Calcium increases induced by single action potentials in rat visual cortex layer II/III pyramidal neurons were shown to be augmented by muscarinic acetylcholine receptor (mAchR) stimulation. This augmentation was drastically reduced by intracellular injection of heparin but not ruthenium red, therefore involving inositol-1,4,5-trisphosphate (IP3)-sensitive rather than ryanodine-sensitive calcium stores. Only the calcium increase induced by the second or later spike of a spike train, but not that induced by the first spike, was augmented, indicating the requirement of both spike-induced calcium increase and mAchR activation. The calcium store depletor thapsigargin abolished this augmentation use-dependently. These findings suggest a neocortical occurrence of calcium-induced calcium release from IP3-sensitive calcium stores that have been sensitized beforehand by IP3 through mAchR-mediated mechanisms.

Action Potentials↗

PubMed2000

Effect of atypical antipsychotics on phencyclidine-induced expression of arc in rat brain.

The effect of atypical antipsychotics on the immediate-early gene, arc (activity-regulated cytoskeleton-associated gene), expression was investigated in phencyclidine (PCP)-treated rats using RT-PCR. Administration of PCP (10 mg/kg) increased arc mRNA levels in the prefrontal cortex, nucleus accumbens and posterior cingulate cortex. Pretreatment with clozapine (20 mg/kg), olanzapine (10 mg/kg) and risperidone (2 mg/kg), but not haloperidol (2 mg/kg), prevented PCP-induced arc expression in the prefrontal cortex and nucleus accumbens. Pretreatment of haloperidol increased the striatal arc mRNA levels. Clozapine, olanzapine and haloperidol inhibited the PCP-induced arc expression in the posterior cingulate cortex. These results suggest that the effects of antipsychotic drugs on PCP-induced arc expression in the prefrontal cortex and nucleus accumbens are useful for distinguishing atypical antipsychotic properties of the drugs.

Animals↗

PubMed2000

Increased D-type cyclin expression together with decreased cdc2 activity confers megakaryocytic differentiation of a human thrombopoietin-dependent hematopoietic cell line.

At the late phase of megakaryocytopoiesis, megakaryocytes undergo endomitosis, which is characterized by DNA replication without cell division. Although a number of cell cycle regulatory molecules have been identified, the precise roles of these molecules in megakaryocytic endomitosis are largely unknown. In a human interleukin-3-dependent cell line transfected with the thrombopoietin (TPO) receptor c-mpl (F-36P-mpl), either treatment with TPO or the overexpression of activated ras (Ha-Ras(G12V)) induced megakaryocytic maturation with polyploid formation. We found that TPO stimulation or Ha-Ras(G12V) expression led to up-regulation of cyclin D1, cyclin D2, and cyclin D3 expression. In addition, expression levels of cyclin A and cyclin B were reduced during the total course of both TPO- and Ha-Ras(G12V)-induced megakaryocytic differentiation, thereby leading to decreased cdc2 kinase activity. Neither the induced expression of cyclin D1, cyclin D2, or cyclin D3 nor the expression of a dominant negative form of cdc2 alone could induce megakaryocytic differentiation of F-36P-mpl cells. In contrast, overexpression of dominant negative cdc2 together with cyclin D1, cyclin D2, or cyclin D3 facilitated megakaryocytic differentiation in the absence of TPO. These results suggest that both D-type cyclin expression and decreased cdc2 kinase activity may participate in megakaryocytic differentiation.

CDC2 Protein Kinase↗