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M Ohno

Publications and source records attributed to M Ohno.

At least 199 records · Page 11Linked to original sources
PubMed1996

Interactive processing between glutamatergic and cholinergic systems involved in inhibitory avoidance learning of rats.

Rats were tested with a one-trial inhibitory avoidance paradigm in which the latency to enter the shock compartment served as a measure of memory retention. Pretraining administration of the non competitive NMDA receptor antagonist MK-801 (0.3 mg/kg i.p.) significantly reduced the response latency during the retention test given 24 h after rats received a step-through inhibitory avoidance training. MK-801 at 0.1 mg/kg did not affect the retention latency. The muscarinic receptor antagonist scopolamine (1.0 mg/kg i.p.) also interfered with the inhibitory avoidance response in the retention test when administered before the training trial. The lower dose of 0.3 mg/kg scopolamine, which by itself was ineffective, significantly impaired inhibitory avoidance learning when administered simultaneously with the behaviorally subthreshold dose of 0.1 mg/kg MK-801 before the training trial. These results suggest that interactive mechanisms regulated by concurrent activation of NMDA and muscarinic receptors are involved in learning processes of inhibitory avoidance performance in rats.

Animals↗
PubMed1996

Characterization of a ligand for receptor protein-tyrosine kinase HTK expressed in immature hematopoietic cells.

HTK is a receptor tyrosine kinase that belongs to the Eph subfamily. An extensive screening using BIAcore system revealed that a colon cancer cell line, C-1, expressed the ligand for HTK. From the conditioned medium of C-1 cells, a soluble form of ligand was purified by receptor affinity chromatography, and the isolation of full-length cDNA revealed that this ligand is identical to the human HTK ligand (HTKL) previously reported. HTK receptor tyrosine phosphorylation was induced by membrane-bound or clustered soluble HTKL but not by unclustered soluble HTKL, indicating that HTKL requires cell-to-cell interaction for receptor activation. Binding analysis demonstrated that HTKL binds to HTK with a much higher affinity (Kd: 1.23 nM) than the other transmembrane-type ligand for Eph family, LERK-2/ELKL (Kd: 135 nM). The expression of HTK in cord blood cells was upregulated after the culture in the presence of stem cell factor. Clustered soluble HTKL stimulated the proliferation of sorted HTK+ cord blood cells and a hematopoietic cell line, UT-7/EPO from which HTK was isolated. These findings suggest the involvement of HTK-HTKL system in the proliferation of HTK+ hematopoietic progenitor cells in the hematopoietic environment.

Amino Acid Sequence↗
PubMed1996

Accelerated evolution of Trimeresurus okinavensis venom gland phospholipase A2 isozyme-encoding genes.

Three Trimeresurus okinavensis (To; himehabu snake, Crotalinae) venom gland phospholipase A2 (PLA2) isozymeencoding genes, gPLA2-o1, gPLA2-o2 and gPLA2-o3, were isolated from its genomic DNA library. The nucleotide (nt) sequence analysis revealed that two of the three genes (gPLA2-o2 and gPLA2-o3) occasionally have been converted to inactivated genes by introduction of one base insertion or substitution. It was confirmed from Southern blot analysis that the To haploid genome contains only three venom gland PLA2 isozyme genes herein isolated. Comparison of these genes showed that nonsynonymous nt substitutions have occurred more frequently than synonymous nt substitutions in the protein-coding regions, except for the signal-peptide coding domain, implying that To venom gland PLA2 isozyme genes have evolved via accelerated evolution. Such an evolutionary feature of To venom gland PLA2 isozyme genes proves the general universality of accelerated evolution previously drawn for venom gland PLA2 isozyme genes of other crotalinae snakes. The variability in the mature protein-coding regions of three To venom gland PLA2 isozyme genes appears to have been brought about by natural selection for point mutations.

Amino Acid Sequence↗
PubMed1996

Beta-adrenergic dysfunction exacerbates impairment of working memory induced by hippocampal NMDA receptor blockade in rats.

To clarify the interactions between hippocampal glutamatergic and adrenergic systems in the working memory function of rats, the effects of hippocampal NMDA receptor blockade combined with noradrenaline depletion or alpha- and beta-adrenoceptor blockade on this behavior were examined with a three-panel runway task. Intrahippocampal administration of the potent and competitive NMDA receptor antagonist (+/-)-3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP) at a dose of 32 ng/side significantly increased the number of errors (attempts to pass through two incorrect panels of the three panels gates at four choice points) in the working memory task, whereas the 3.2 ng/side dose of CPP did not affect working memory errors. Administration of the noradrenergic neurotoxin N-(2-chloroethyl)-N-ethyl-2 -bromobenzylamine (DSP-4) at 50 mg/kg i.p. produced marked reductions in hippocampal and cortical noradrenaline contents, but it had no effect on working memory errors. Intrahippocampal administration of 3.2 ng/side CPP, the behaviorally ineffective dose in intact rats, significantly increased the number of working memory errors in the noradrenaline-depleted rats. The alpha-adrenoceptor antagonist phentolamine (3.2 mg/kg i.p.) did not affect working memory errors whether administered alone or in combination with intrahippocampal CPP (3.2 ng/side). The beta-adrenoceptor antagonist propranolol (10 mg/kg i.p.) also had no effect on working memory errors. However, propranolol (10 mg/kg) produced a significant increase in working memory errors when administered together with intrahippocampal CPP (3.2 ng/side). These results suggest that hippocampal NMDA/beta-adrenergic interactions are involved in neural processes mediating working memory function of rats.

Adrenergic alpha-Antagonists↗
PubMed1996

Antianginal effects of FR144420, a novel slow nitric oxide-releasing agent.

The aim of this study was to compare the antianginal effects of two compounds that release nitric oxide (NO) spontaneously, i.e. (+/-)-N-[(E)-4-ethyl-3-[(Z-hydroxyimino]-5-nitro-3-hexenyl] -3-pyridinecarboxamide (FR144420) and (+/-)-(E)-ethyl-2-[(E)-hydroxyimino] -5-nitro-3-hexenamide (FK409), in two different rat models of coronary vasospasm. In the rat methacholine-induced coronary vasospasm model, FR144420 suppressed the elevation of the ST segment dose dependently and significantly at 1.0 mg/kg, i.d. 185 min after its administration. FK409 suppressed the ST elevation only 5 min after its administration at 1.0 mg/kg, i.d. FR144420 and FK409 significantly decreased mean blood pressure at all doses tested only 5 min after their intraduodenal administration, but did not change heart rate at any time. Although the suppression of the ST elevation by FK409 had the same duration as its hypotensive effect, the FR144420-induced suppression of the ST elevation lasted longer than its hypotensive effect. In the rat vasopressin-induced coronary vasospasm model, FR144420 (32 mg/kg) significantly inhibited the depression of the ST segment both 60 min and 120 min after oral administration, whereas FK409 (32 mg/kg) significantly inhibited this ST depression only 60 min after oral administration. These data suggest that FR144420 inhibits coronary vasospasm for longer than FK409 does and particularly shows more prolonged antianginal effects than hypotensive effects in the methacholine-induced coronary vasospasm model. Thus FR144420 is expected to be a useful NO releaser for investigating the in vivo actions of NO.

Animals↗
PubMed1996

Serum carcinoembryonic antigen levels and proliferating cell nuclear antigen labeling index for patients with colorectal carcinoma. Correlation with tumor progression and survival.

BACKGROUND: Clinicopathologic variables, carcinoembryonic antigen (CEA), nuclear DNA ploidy, and proliferating cell nuclear antigen labeling index (PCNA LI) have been studied for their effect on patients with various types of cancer. METHODS: Thirteen clinicopathologic variables, preoperative serum CEA levels, PCNA LI, DNA ploidy patterns, and survival were studied for 57 colorectal carcinoma patients, and the mutual relation between these variables, tumor progression, and survival were analyzed. RESULTS: Seven variable including undifferentiated adenocarcinoma, deep invasion, lymphatic and venous invasion, node metastasis, liver metastasis, and advanced stages were significantly greater for patients with positive CEA ( > 5.0 ng/mL) than for patients with negative CEA ( < 5.0 ng/ML). Three variables including deep invasion, liver metastasis, and advanced stages were significantly greater for patients with high PCNA LI ( > 49.4%) than for patients with low PCNA LI ( < 49.4%). No significant difference was observed in any of the variable for patients with DNA diploid or aneuploid cancer. A close relationship representing a formula, Y (log CEA, ng/mL) = 0.026X (PCNA LI, %) - 0.478, was found between CEA level and PCNA LI: Survival curves for patients with negative CEA and low PCNA LI were significantly greater than those for patients with positive CEA and high PCNA LI: Survival curves were significantly greater for patients with positive CEA and low PCNA LI than for patients with positive CEA and high PCNA LI: However, no significant difference was observed in the survival curves of patients with different DNA ploidy patterns. CONCLUSIONS: Serum CEA and PCNA LI for cancer patients is useful in the evaluation of tumor progression and in prognosis. Analysis of DNA ploidy appears to be unavailable for this evaluation.

Adult↗
PubMed1996

A human RNA helicase-like protein, HRH1, facilitates nuclear export of spliced mRNA by releasing the RNA from the spliceosome.

Because the nuclear export of mRNA occurs only after the splicing reaction is completed, intron-containing pre-mRNA does not normally appear in the cytoplasm. As a mechanism to secure this, intron-containing RNA is retained in the nucleus via formation of the spliceosome. Therefore, the process of releasing spliced mRNA from the spliceosome after completion of splicing is an essential step for triggering the nuclear export of the spliced mRNA. In budding yeast, RNA helicase-like protein Prp22 is implicated in this process. Here we demonstrate the function of HRH1, a human protein homologous to Prp22, in mammalian cells using dominant-negative HRH1++ mutants (dn-HRH1). dn-HRH1 protein stalls on the spliceosome and prevents release of the spliced RNA from the spliceosome in vitro. Expression of dn-HRH1 in mammalian cells leads to inhibition of splicing and to extensive nuclear export of unspliced pre-mRNA, probably because of the incapability of recycling spliceosome components that normally retain the pre-mRNA in the nucleus. The arginine/serine-rich domain (RS domain) of HRH1, which is missing in Prp22, confers a nuclear localization signal, and appears to facilitate the interaction of HRH1 with the spliceosome. This is the first report on a bona fide mammalian homolog of yeast Prp splicing factor, and also on a mammalian RNA helicase-like splicing factor.

Amino Acid Sequence↗
PubMed1996

Roles of dopamine D1 receptors in delta 9-tetrahydrocannabinol-induced expression of Fos protein in the rat brain.

We examined whether administration of delta 9-tetrahydrocannabinol (THC) induces the expression of Fos protein or not in the rat brain. A single administration of 3.2 and 10 mg/kg THC produced a dose-dependent and significant increase in Fos-immunoreactive cells in the striatum, particularly in its dorsomedial portions. The peak increase was reached 2 h after THC treatment and was absent at 8 h. Fos induction was also observed in the nucleus accumbens after administration of 10 mg/kg THC. However, in the globus pallidus, hippocampus and substantia nigra pars reticula, abundant in cannabinoid receptors, there were few or no Fos-immunoreactive cells induced by 10 mg/kg THC. SCH-23390, a selective dopamine D1 receptor antagonist, at 0.32 mg/kg produced a significant block of the effects of THC on Fos expression in the striatum and the nucleus accumbens. Administration of 100 mg/kg (-)-sulpiride, a selective dopamine D2 receptor antagonist, failed to block the effect of THC on Fos expression in both the striatum and the nucleus accumbens. These findings indicate that THC induces the expression of Fos protein and that this expression is mediated at least by dopamine D1 receptors.

Animals↗
PubMed1996

Concurrent blockade of hippocampal metabotropic glutamate and N-methyl-D-aspartate receptors disrupts working memory in the rat.

In order to clarify the roles of hippocampal metabotropic glutamate and N-methyl-D-aspartate receptors in working and reference memory performance of rats, the effects of intrahippocampal administration of selective antagonists for both receptors on these behaviours were examined with a three-panel runway task. In the working memory task, the potent and competitive N-methyl-D-aspartate receptor antagonist, (+/-)-3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP), significantly increased the number of errors (attempts to pass through two incorrect panels of the three panel-gates at four choice points), when injected bilaterally at 10 and 32 ng/side into the dorsal hippocampus. Intrahippocampal injection of CPP at a dose of 3.2 ng/side had no effect on the number of working memory errors. The metabotropic glutamate receptor antagonist, (+)-alpha-methyl-4-carboxyphenylglycine (+)-(MCPG), injected into the hippocampus at doses up to 3.2 micrograms/side, did not significantly affect the number of working memory errors. Combined administration of (+)-MCPG (3.2 micrograms/side) and CPP (3.2 ng/side) into the hippocampus, neither of which had an individual effect on errors, significantly increased the number of working memory errors. However, intrahippocampal administration of the relatively inactive isomer, (-)-MCPG, at 3.2 micrograms/side did not affect working memory errors, whether given independently or concurrently with the behaviourally ineffective dose of CPP (3.2 ng/side). In the reference memory task, intrahippocampal injection of CPP at doses up to 32 ng/side had no effect on the number of errors. Intrahippocampal (+)-MCPG at doses up to 3.2 micrograms/side did not affect the number of reference memory errors, whether administered alone or together with 3.2 ng/side of CPP. These results indicate that blockade of hippocampal metabotropic glutamate receptors aggravates impairment of working memory resulting from deficiency of N-methyl-D-aspartate receptor-mediated glutamatergic neurotransmission, suggesting that mechanisms regulated by co-activation of hippocampal metabotropic glutamate and N-methyl-D-aspartate receptors are involved in working memory performance of rats.

Animals↗
PubMed1996

Ischemic tolerance to memory impairment associated with hippocampal neuronal damage after transient cerebral ischemia in rats.

When rats were trained preoperatively with a three-panel runway task and were then exposed to 10-min ischemia by the method of 4-vessel occlusion, they showed no increase in the number of errors (attempts to pass through two incorrect panels of the three panel-gates at four choice points), having normal retention of memory performance learned before the ischemic insult. Next, we investigated the abilities of ischemic rats to acquire the three-panel runway task and to learn a subsequent reversal task, where the correct panel-gate locations were changed. Rats with 5-min ischemia exhibited performance as good as that of control rats, but rats exposed to 10- and 20-min ischemia showed more errors than control rats during 10 acquisition sessions and 5 subsequent reversal sessions, each of which (consisting of 6 trials) was given once a day. Marked neuronal degeneration was observed in the hippocampal CA1 sector from the rats with 10- and 20-min ischemia. Exposure to sublethal 5-min ischemia followed by 10-min ischemia at a 2-h interval had no effect on either the memory impairment during acquisition and reversal tests or the hippocampal CA1 damage. When rats were exposed to 5-min ischemia 2 days before lethal 10-min ischemia, they showed acquisition and subsequent reversal learning as good as that of control rats. Preconditioning with sublethal 5-min ischemia followed by 2 days of reperfusion also prevented the neuronal destruction of the hippocampal CA1 sector induced by 10-min ischemia. These findings suggest that postischemic hippocampal CA1 neuronal damage does not affect retention of spatial memory acquired before ischemia, but produces a significant impairment of acquisition and subsequent reversal learning. The present results also demonstrate that preconditioning with sublethal ischemia can develop tolerance to subsequent lethal ischemia to prevent the learning impairment related to the hippocampal CA1 neuronal damage.

Analysis of Variance↗
PubMed1996

ZTTA, a postproline cleaving enzyme inhibitor, improves cerebral ischemia-induced deficits in a three-panel runway task in rats.

We investigated the effect of N-benzyloxycarbonyl-thioprolyl-thioprolinal-dimethylaceta l (ZTTA), a novel postproline cleaving enzyme (prolyl endopeptidase, PPCE) inhibitor, on the in vitro activity of rat brain PPCE and memory impairment induced by cerebral ischemia. ZTTA noncompetitively inhibited rat brain PPCE (ki = 2.9 microM). Cerebral ischemia for 5 min increased the number of errors in a working memory task with a three-panel runway paradigm. ZTTA at 6 mg/kg, administered immediately after blood flow reperfusion, significantly reduced the increase in working memory errors expected to occur 24 h after 5 min of ischemia. The antiamnesic action of ZTTA may be ascribable to a neuroprotective effect on the central nervous system due to some neuropeptides that are substrates of PPCE in the brain.

Animals↗
PubMed1996

An external reference for in vivo quantification of 1H spectroscopy.

We studied the validity of using an external signal reference for absolute quantification of brain metabolites. We chose cyclohexane as the signal standard. An expression of a perturbed magnetic field in diamagnetic cylindrical materials was first obtained by using a method of equivalent magnetic charges. Broadening of the spectrum in this perturbed magnetic field was then computed. We performed phantom studies. We measured the full width at half maximum (FWHM) of the cyclohexane and the signal ratio (SR) between water and cyclohexane in spheres of two different diameters, and in cylinders of various diameters and lengths. The observed FWHM and SR in the small cylinders had large variations in distribution, whereas those in the spheres and in the large cylinders had small variations. Specifically, in a cylinder with a diameter > 5 cm and with a length > 10 cm, the variation of the SR was < 1.0%. The diameter and the length of the cylinder preferably are large when a cylindrical material is adopted as an external signal standard.

Brain↗
PubMed1996

Effects of Fe(2+), Zn(2+), Cu(2+) and Se(4+) on the synthesis and catabolism of prostaglandins in rabbit gastric antral mucosa.

Effects of Fe(2+), Zn(2+), Cu(2+) and Se(4+) on the synthesis and catabolism of prostaglandins (PGs) in rabbit gastric antral mucosa were examined. Fe(2+) inhibited the cyclooxygenase activity in the microsomal fraction. Zn(2+) suppressed the endoperoxide E(2) isomerase activity in the microsomal fraction and the 15-hydroxy PG dehydrogenase (PGDH) activity in the cytosolic fraction. Cu(2+) stimulated the cyclooxygenase activity, inhibited the PGDH activity and possibly induced non-enzymatic reduction of PGG(2) or PGH(2) to PGF(2 alpha) Se(4+) possibly induced the non-enzymatic reduction of PGG(2) or PGH(2) to PGF(2 alpha), as well as Cu(2+). These results suggest that Fe(2+), Zn(2+), Cu(2+) and Se(4+) can be modulators of the gastric antral mucosal PG levels by affecting the PG synthesizing enzymes, PG catabolizing enzyme and/or non-enzymatic reduction of PGG or PGH to PGF.

Animals↗
PubMed1996

Specificity of glossopharyngeal nerve responses to astringent compounds in Xenopus.

Astringent compounds were applied to oral epithelium of the clawed toad, Xenopus laevis, and rapidly rising and highly sensitive responses could be recorded from the whole glossopharyngeal nerve, but not at all from the trigeminal nerve. The response to 10 mM tannic acid decreased progressively with repetitive application. These responses to tannic acid, however, recovered completely by treating with chemicals capable of forming strong hydrogen and hydrophobic bonds. These chemical bondings are generally recognized as a model for polyphenol (tannin)-protein interactions based on physico-chemical measurements in vitro. The high affinities of these chemicals for tannic acid may be effective in releasing both bonds in the interaction of tannic acid with the receptor molecules. Our results provide in vivo evidence for this model.

Animals↗
PubMed1996

Chymotrypsin inhibition induced by side chain-side chain intramolecular CH/pi interaction in D-Thr-L-Phe benzylamide.

The dipeptide benzyl amide H-D-Thr-Phe-NH-CH2-C6H5 was found to inhibit chymotrypsin strongly (K1 = 4.5 x 10(-6) M) in a competitive manner. When a series of phenyl amides H-D-Thr-Phe-NH-(CH2)n-C6H5 (n = 0-4) were tested, inhibitory potency peaked at n = 1 (benzyl amide). Incorporation of a methyl group into the benzyl methylene resulted in formation of stereoisomers, H-D-Thr-Phe-NH-(R or S)-CH(CH3)-C6H5, with considerably different inhibitory potencies. The R-isomer was as active as the benzyl amide, while the S-isomer was about 30-fold less active than the benzyl amide. Furthermore, when a fluorine atom was introduced into the para-position of the amide-benzyl group, the resulting H-D-Thr-Phe-NH-CH2-C6H4(p-F) showed considerably enhanced inhibitory activity (about 5-fold, K1 = 9.1 x 10(-7) M). In conformational analysis by 400 mHz 1H-NMR, all dipeptides having D-Thr-Phe backbone structure showed large upfield shifts of D-Thr-beta OH (shifts in ppm, 0.09-0.17), D-Thr-beta CH (0.23-0.32), and D-Thr-gamma CH3 (0.38-0.53), indicating the presence of shielding effects from the benzene ring. In addition, NOE enhancements between the D-Thr-gamma CH3 and Phe-phenyl groups were evidenced by measurements of two-dimensional NOESY spectra and NOE difference spectra. These observations demonstrated the spatial proximity of these side chains, which is due to side chain-side chain CH/pi interaction. All these results support the idea that the amide-benzyl group binds at the chymotrypsin S1 site, while the hydrophobic core with CH/pi interaction binds at the S2 or S1' site.

Chymotrypsin↗
PubMed1996

Altered ventricular and myocyte response to angiotensin II in pacing-induced heart failure.

Alterations in the cardiac response to angiotensin II (Ang II) may contribute to the functional impairment in tachycardia-induced heart failure (congestive heart failure [CHF]). Accordingly, we studied the response to Ang II in eight conscious instrumented dogs before and after inducing CHF. Left ventricular (LV) performance was assessed by measuring LV pressure and LV volume. Isolated myocyte function was evaluated using computer-assessed videomicroscopy. In conscious animals before CHF, Ang II produced a load-dependent slowing of the time constant of LV relaxation (tau) and did not depress intact LV contractile function. After CHF, although Ang II produced a similar increase in LV systolic pressure, the increases in LV diastolic pressure and time constant tau were much greater, and contractile performance was depressed. These changes persisted when the elevation of end-systolic pressure was prevented by nitroprusside. Similar changes were also present after autonomic blockade. In isolated myocytes, before CHF, Ang II (10(-6) mol/L) produced a slight positive inotropic effect. In contrast, after CHF, Ang II produced a negative inotropic effect and slowed the rate of relengthening. The effects in the intact LV and myocytes were reversed by an Ang II AT1 receptor blocker (losartan). We conclude that pacing-induced CHF alters the LV and myocyte response to Ang II, so that Ang II produces direct depressions in intact LV contraction, relaxation, and filling and exacerbates myocyte contractile dysfunction. These effects are mediated through the activation of AT1 receptors.

Angiotensin II↗
PubMed1996

Purification and characterization of a cellulase from the giant snail Achatina fulica.

A cellulase with activity as a cellobiohydrolase [EC3.2.1.91] was purified from the intestinal juice of the giant snail Achatina fulica. The enzyme (M(r) = about 23,000, and pI = about 5.3) also had a weak beta-glucosidase activity. The amino acid sequence of the N-terminal 20 amino acids was analyzed, and no similarity was noted for sequences of other known cellobiohydrolases.

Amino Acid Sequence↗