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Biomedical subjects

K Wada

Publications and source records attributed to K Wada.

At least 343 records · Page 19Linked to original sources
PubMed1996

Repeated hyperbaric oxygen induces ischemic tolerance in gerbil hippocampus.

Hyperbaric oxygen (HBO; 100% oxygen at 2 atmospheres absolute) was administered for 1 h to male Mongolian gerbils either for a single session or every other day for five sessions. Two days after HBO pretreatment, the gerbils were subjected to 5 min of forebrain ischemia by occlusion of both common carotid arteries under anesthesia. Seven days after recirculation, neuronal density per 1-mm length of the CA1 sector in the hippocampus was significantly better preserved in the five-session HBO pretreatment group (n = 10: 175.7 (47.8/mm, 54.9% of normal) than in the ischemic control group (n = 10: 26.2 (11.6/mm, 8.0% of normal) and in the single-session HBO pretreatment group (n = 7: 37.3 (21.7/mm, 11.4% of normal). Immunohistochemical staining for the 72-kDa heat-shock protein (HSP-72) in the CA1 sector performed 2 days following pretreatment revealed that the five-session HBO pretreatment increased the amount of HSP-72 present compared with that in the ischemic control group and in the single HBO pretreatment group. These results suggest that tolerance against ischemic neuronal damage was induced by repeated HBO pretreatment, which is thought to occur through the induction of HSP-72 synthesis.

Animals↗
PubMed1996

Sensitive determination of nitrotyrosine in human plasma by isocratic high-performance liquid chromatography.

A highly sensitive and simple isocratic high-performance liquid chromatographic method was developed for determination of 3-nitrotyrosine in human plasma with precolumn derivatization with 4-fluoro-7-nitrobenzo-2-oxa-1,3-diazole. The precision of the method was satisfactory (coefficient of variation 4.8%), and the detection limit was established at 0.1 pmol of 3-nitrotyrosine allowing the determination at the level of 6 pmol/ml in human plasma. The recoveries of 3-nitrotyrosine and alpha-methyltyrosine, an internal standard, were 89.3 +/- 7.1 and 85.7 +/- 7.6%, respectively. The 3-nitrotyrosine level was 31 +/- 6 pmol/ml (mean +/- S.D., n = 9) in plasma from healthy volunteers. Since 3-nitrotyrosine is a stable product of peroxynitrite, an oxidant formed by a reaction of nitric oxide and superoxide radicals, the measurement of its plasma concentration may be useful as a marker of nitric oxide-dependent oxidative damage.

4-Chloro-7-nitrobenzofurazan↗
PubMed1996

EAAT4 is a post-synaptic glutamate transporter at Purkinje cell synapses.

To study cellular and subcellular localizations of the glutamate transporter EAAT4, antibody was raised against the N-terminal peptide. On immunoblotting the antibody recognized a band in membrane extracts from the cerebellum, but not from the forebrain. Immunohistochemistry revealed that its distribution was restricted to the cerebellar molecular layer, where the immunoreactivity was observed as numerous punctate stainings. Electron microscopy showed the antibody to label dendritic spines of the Purkinje cells. EAAT4 is, therefore, a Purkinje cell-specific, postsynaptic transporter. Together with dense localization of other transporter subtypes in Bergmann astrocytic membranes, Purkinje cell synapses are thus provided with distinct glutamate transporter subtypes at discrete synaptic elements, which would play important roles in regulating excitability of the Purkinje cells and protecting against excitotoxicity.

ATP-Binding Cassette Transporters↗
PubMed1996

Effect of intranasal administration of thyrotropin-releasing hormone on ataxic gait in staggerer mice.

The ataxia ameliorating effect of an intranasal administration of thyrotropin-releasing hormone (TRH) was examined using normal and ataxic staggerer mutant mice. In the normal mice, the blood TRH level reached the maximum level 5 min after administration and was gradually eliminated during the following 60 min. The antiataxic effects of TRH in the staggerer mice was examined using an open field method. At lower doses, the intranasal administration of TRH in the staggerer mice was examined using an open field method. At lower doses, the intranasal administration of TRH did not exert any evident effect. However, at 3 mg or 4 mg, the fall index (the ratio of the number of falls to the movement score) was significantly decreased for 20 min after the administration. These results show that an intranasal administration of TRH can ameliorate the ataxia in staggerer mice, and may be promising for clinical use in patients with spinocerebellar degeneration.

Animals↗
PubMed1996

Requirement of cell interactions through adhesion molecules in the early phase of T cell development.

We investigated the role of adhesion molecules in T cell development. A large proportion of murine fetal thymus (FT) cells obtained at Day 13 of gestation, which are c-kit+, express the adhesion molecules Pgp-1, VLA-4, LFA-1, and ICAM-1 on their surface at high levels. The expression profiles of these adhesion molecules resemble quite well those on c-kit+ cells in fetal liver (FL). The level of expression of these molecules on FT cells declines with the embryonal age and becomes mostly negative by birth except for LFA-1. In the case of LFA-1, a reincrease of expression levels is seen in newborn mice. The role of these adhesion molecules in T cell development was investigated by adding monoclonal antibodies (mAb) into the FT organ cultures, where T cell development from FT or FL progenitors was induced by coculturing these cells with a deoxyguanosine-treated FT lobe. We found that anti-Pgp-1, anti-LFA-1, and anti-VLA-4 mAb severely inhibited the early phase of T cell development from FL progenitors. On the other hand, the suppressive effect of these mAb on the T cell development from FT progenitors was only slight, if any. These findings strongly suggest that interactions with elements in the thymic microenvironment through Pgp-1, LFA-1, and VLA-4 are indispensable for prethymic progenitors to develop into T cells.

Animals↗
PubMed1996

Genomic organization, promoter analysis, and chromosomal localization of the gene for the mouse glial high-affinity glutamate transporter Slc1a3.

The mouse gene encoding glial high-affinity, Na+-dependent glutamate transporter Slc1a3 (GluT-1/GLAST) was isolated, and its structural organization was characterized. The gene appeared to exist as a single copy in the mouse genome and comprised 10 exons spanning more than 56 kilobases. The transcription initiation sites were mapped to positions 503, which is the first transcriptional point (defined as +1), 128 (+376), and 64 (+440) basepairs upstream of the 3'-end of exon 1 by primer extension. The 5'-flanking region of the mouse GluT-1 gene had a typical CCAAT box and a GC box but lacked a TATA box. These features of the promoter region were characteristic of housekeeping genes. The fusion plasmids containing approximately 4 kb of the 5'-flanking region (-3830 to +450) and the firefly luciferase gene induced a significant luciferase activity when transfected into COS-1 cells. Distal deletion of the 5'-flanking region, leaving 619 bp (-169 to +450), resulted in a marked decrease in luciferase activity in COS-1 cells, suggesting that a CCAAT box, which was positioned at -200, is necessary for the expression of this gene. In situ hybridization localized this gene to mouse chromosome 15A2. These structural features will lead to a better understanding of the regulatory mechanism of the expression of the GluT-1 gene by ischemia and will also provide a basis for future evolutionary comparisons with other neurotransmitter transporters.

ATP-Binding Cassette Transporters↗
PubMed1996

Dynamic changes in expression of glutamate transporter mRNAs in developing brain.

Developmental changes in gene expression for three glutamate transporter subtypes in the mouse brain were analysed by in situ hybridization. During embryonic stages, GluT-1 and GLT-1 mRNAs were expressed at high levels in the ventricular zone, whereas EAAC1 mRNA was not detected in the zone. In the mantle zone, transcription levels of three transporter mRNAs were low during embryonic stages, and these levels, especially those of the GluT-1 and GLT-1 mRNAs, displayed remarkable increases postnatally to reach maximal levels at 14 days of age. These dynamic developmental regulations suggest that the glutamate transporter not only regulates the excitatory synaptic transmission at mature stages, but might also be intimately involved in the brain development.

ATP-Binding Cassette Transporters↗
PubMed1996

Effect of cystathionine as a scavenger of superoxide generated from human leukocytes or derived from xanthine oxidase in vitro.

We studied the direct effects of cystathionine on human leukocyte-generated or xanthine-xanthine oxidase-derived superoxide radicals in vitro. Washed leukocyte suspensions (10(6) cells/ml) prepared from healthy male volunteers were stimulated with phorbol myristate acetate (1 mu M) or opsonized zymosan (1 mg/ml) to generate superoxide radicals, which were measured with a 2-methyl-6-[p-methoxyphenyl]-3,7-dihydroimidazo[1,2-alpha] pyrazin-3-one hydrochloride (MCLA)-chemiluminescence method. Cystathionine (30 mu M to 10 mM) significantly reduced superoxide radical-dependent chemiluminescence in the leukocyte system in a concentration-dependent manner. In addition, in the two different methods of determination of superoxide radicals (MCLA chemiluminescence and nitroblue tetrazolium reduction), cystathionine significantly scavenged the superoxide radicals derived from the xanthine-xanthine oxidase system. However, cystathionine did not inhibit the activity of xanthine oxidase during superoxide generation. On the other hand, cystathionine did not show a scavenging effect against hydroxyl radicals derived from Fe2+ -H2O2 on the erythrocyte membrane. These results indicate that cystathionine itself may possess a scavenging function against superoxide radicals rather than against hydroxyl radicals in vitro.

Cystathionine↗
PubMed1996

Codon usage tabulated from the international DNA sequence databases.

Codon usage in 87 602 genes has been calculated using the nucleotide sequence data obtained from the GenBank Genetic Sequence Data Bank (Release 90.0; September 1995). The database is called the CUTG Database; the complete form of the database can be obtained by anonymous ftp from DDBJ and a part of the database, which lists the frequency of codon use in each organism, is made searchable through our World Wide Web server.

Base Sequence↗
PubMed1996

Release of taurine and its effects on release of neurotransmitter amino acids in rat cerebral cortex.

Taurine has been postulated to function as a neurotransmitter or neuromodulator. The possibility of depolarization-evoked release of taurine from nerve terminals, and the effects of taurine on release of endogenous glutamate (Glu), aspartate (Asp), and gamma-aminobutyrate (GABA) were examined using a superfusion of crude synaptosomes prepared from rat cerebral cortex. Taurine contents in cerebral cortex and its synaptosomes were 31.7 and 25.2 nmol/mg protein, respectively. Although the basal rate of taurine release was 35.3 pmol/min/mg protein of synaptosomes (second highest releasing rate), the 2-min stimulation with KCl (30 mM) evoked only a 1.3-fold increase in release of taurine (47.3 pmol/min/mg). The increase was largely Ca(2+)-dependent. The addition of taurine to the perfusion medium significantly reduced the depolarization-evoked increases in Glu, Asp, and GABA release. The taurine-induced reduction in GABA release was attenuated by phaclofen, a GABAB antagonist, but not by bicuculline, a GABAA antagonist. However, these antagonists did not block the effects on Glu and Asp release. These data suggest that taurine may be only partly released from nerve terminals by depolarization in the cerebral cortex, but that taurine may act upon nerve terminals to regulate the release of excitatory and inhibitory amino acid transmitters.

Animals↗
PubMed1996

Frequent loss in chromosome 8p loci in liver cirrhosis accompanying hepatocellular carcinoma.

Most hepatocellular carcinoma (HCC) is preceded by liver cirrhosis, but the genetic changes involved in cirrhosis are not well understood. We therefore studied loss of heterozygosity (LOH) in cirrhotic and neoplastic foci in livers of 14 patients with HCC. The samples, microdissected from paraffin-embedded tissues, were analyzed using a polymerase-chain-reaction-based assay for dinucleotide repeat polymorphisms on 8p. Of the 14 cases, 13 showed constitutional heterozygosity for the microsatellite markers. In 7 (54%) of these 13 informative cases, LOH was detected in the primary HCC and, in these 7 doubly informative (informative and LOH-positive in primary HCC) cases, LOH was found in 16 (70%) of 23 liver cirrhotic foci. The pattern of 8p allelic loss was identical in each doubly informative tumor; however, some of the liver cirrhotic foci harbored an 8p loss identical to that seen in the primary HCC, some harbored a different 8p loss, and some did not harbor any 8p loss. The remaining 6 cases without LOH on 8p in HCC showed no 8p loss in any cirrhotic foci. Presumably HCC could develop from cirrhotic cells harboring 8p loss.

Adult↗
PubMed1996

Effects of a small dose of triazolam on P300 and resting EEG.

The aim of the present study was to clarify whether cognitive impairments caused by benzodiazepines (BDZs) are a consequence of their specific direct effects on cognitive function or whether they are explained as secondary effects of increased sleepiness. Ten healthy men (mean age, 33.9 years) participated in two experimental sessions in a randomized cross-over, double-blind study: in one session subjects were given a placebo and in the other they were given 0.125 mg triazolam (TRZ). Each experimental session was conducted on 1 day. After a pre-drug EEG recording and an event-related potential (ERP) recording, under an oddball paradigm, subjects took the TRZ or placebo orally at 1000 hours. Thereafter, EEG and ERP recording sessions, following the same procedure as the pre-drug sessions, were conducted at 1, 2, 4, 6 and 8 h after drug administration. The EEG and ERP recordings from Cz and Pz referred to the bilaterally linked ear electrodes were used. We found that P300 latency was significantly prolonged in TRZ condition at 2 h (Pz) and 4 h (Cz and Pz) after TRZ, and that the P300 amplitude was significantly reduced at 2 h (Cz and Pz) and 4 h (Pz) after TRZ, compared to the same times after placebo. The absolute power values for the theta (4-7 Hz), alpha 1 (8-9 Hz), and alpha 2 (10-12 Hz) bands did not differ at any measurement time between the treatments. Only the beta band (13-19 Hz) power value was significantly elevated after the TRZ administration (versus placebo). No significant sedative effects were detected in subjective measurements. These results indicate that a single oral dose of 0.125 mg TRZ caused cortical changes without distinct general sedation or subjective sleepiness.

Adult↗
PubMed1996

Effects of growth factors and gut hormones on proliferation of primary cultured gastric mucous cells of guinea pig.

Almost completely homogenous gastric mucous epithelial cells of guinea pigs were grown to confluence in the presence of 10% fetal calf serum (FCS). FCS, epidermal growth factor (EGF), and insulin significantly increased 5-bromo-2'-deoxyuridine (BrdU) uptake by the cells and EGF together with insulin increased the cells' [3H] thymidine uptake. Basic fibroblast growth factor (bFGF) enhanced EGF-induced DNA synthesis by the cells, but vasoactive intestinal peptide (VIP), secretin, prostaglandin E2 (PGE2), and dibutyryl cyclic AMP (dbcAMP) neither induced DNA synthesis nor enhanced the effect of EGF on DNA synthesis by the cells. Gastrin, cholecystokinin-octapeptide (CCK8), and carbamylcholine chloride (CCh) also did not enhance the effect of EGF on DNA synthesis. 125I-EGF, 125I-bFGF, and 125I-gastrin binding to the gastric mucous cells revealed the presence of high-affinity receptors for EGF and bFGF, but not for gastrin. Northern blot analysis showed the expression of EGF receptor mRNA, but not gastrin receptor mRNA. These results suggest that EGF, insulin, and bFGF may cooperatively regulate gastric mucous cell growth, but that gastrin and other gastrointestinal hormones do not have a direct stimulatory effect on mucous cell growth in the guinea pig.

Animals↗
PubMed1996

Intraventricular administration of estradiol modulates rat prolactin secretion and synthesis.

The effect of estradiol (E2) on rat tuberoinfundibular dopaminergic (TIDA) neurons was examined in vivo, employing chronic intraventricular (i.c.v.) infusion technique using an osmotic mini-pump. The activity of TIDA neurons was assessed by the release and synthesis of prolactin (PRL) in the rat pituitary gland and by the changes in the 3, 4-dihydroxyphenylacetic acid (DOPAC) and dopamine (DA) levels and in the DOPAC/DA ratio in the rat hypothalamus. We also examined the [3H] E2 binding in the rat hypothalamus. Ovariectomized female Wistar rats with E2 replacement were treated with daily i.c.v. infusion of 1 microM of E2 or saline vehicle for 1, 3, and 7 days using the Alzet osmotic mini-pump and brain infusion kit. At 1 day of i.c.v. infusion of E2, the serum PRL level was significantly decreased compared with that in the vehicle group. Northern blot analysis of the total RNA isolated from the pituitary glands demonstrated a decrease in the PRL gene transcript level in the E2 group. At 3 days of E2 treatment, however, the serum PRL level was significantly increased compared with that of the vehicle-injected group and Northern blot analysis also demonstrated that the PRL gene transcript level was increased in the E2 group. At 7 days of E2 administration, there were no significant differences between the E2 and vehicle groups in either serum PRL or PRL gene transcript levels. There was a significant increase in the DOPAC/DA ratio after 1 day in the E2 group. However, no significant effects of E2 on this ratio were observed at 3 and 7 days of treatment. The DOPAC concentration in the E2 group was significantly increased at day 1 and significantly decreased at day 3, compared with that of the respective time in vehicle group. At day 7 there was no significant change in DOPAC concentration in either groups. The DA concentrations in the hypothalamus was not changed on any day in either group. Specific [3H] E2 binding was observed in the rat hypothalamus. These data suggest that E2 may have a biphasic effect on the accumulation of PRL gene transcripts and on the PRL secretion in the rat pituitary by first stimulating and then inhibiting the TIDA neuronal activity.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1996

A chemiluminescent detection of superoxide radical produced by adherent leucocytes to the subendothelium following thrombolysis: studies with a photochemically induced thrombosis model in the guinea pig femoral artery.

Reocclusion following thrombolysis is a major limitation of thrombolytic therapy with recombinant tissue-type plasminogen activator (rt-PA) because denuded vessel wall exposed to blood following thrombolysis is a favourable surface for platelet and leucocyte deposition. We have applied a chemiluminescence technique to detect superoxide radical (0(-2)) produced by leucocytes adherent to the femoral artery 24 h after photochemically induced thrombogenesis in the guinea pig in vivo and subsequent thrombolysis by rt-PA. Intravenous administration of MCLA, a specific chemiluminescence reagent for detecting O(-2), markedly increased photon emission. the photon emission was markedly potentiated by phorbol myristate acetate and was suppressed by superoxide dismutase. Reocclusion 24 h after rt-PA induced thrombolysis was observed in 10 of 16 animals. Histological observations revealed extensive polymorphonuclear leucocytes adherent to the vessel wall at the site of thrombogenesis and thrombolysis. A higher level of 0(-2) could be detected from the arteries in which thrombolysis was induced compared with those without thrombolysis. Further, the level 0(-2) detected was greater in reoccluded arteries compared with those in which reflow was established. These observations suggest that 0(-2) is produced by adherent leucocytes at the site of thrombolysis and that leucocytes are involved in reocclusion after thrombolysis.

Animals↗
PubMed1996

A new gastric ulcer model induced by ischemia-reperfusion in the rat: role of leukocytes on ulceration in rat stomach.

A new model of gastric ulcer involving damage to the muscularis mucosae was developed by clamping the celiac artery in rat to induce ischemia-reperfusion (I-R) injury. Although erosions with falling off of the gastric mucosa were observed immediately, 24 and 36 hours after the I-R, gastric ulcers involving the injury of muscularis mucosae were observed in the area of gastric glands at 48 and 72 hours after initiation of injury. Administration of omeprazol, a proton pump inhibitor, or pentoxifylline, an anti-leukocyte drug, just after the initiation of injury significantly decreased the total area of ulcers at 72 hours. A combination of omeprazol and pentoxifylline was more effective than either drug alone. An anti-leukocyte adhesion molecule (anti-CD18 antibody) also showed significant inhibitory effect on the development of ulcers at 72 hours and the infiltration of leukocytes into both submucosa and mucosa. These results indicate that in our model, gastric acid together with leukocytes contribute to the development of ulcers following erosions. This model may be used to investigate the mechanisms of the development of gastric ulcer and evaluate antiulcer drugs in a preclinical setting.

Animals↗
PubMed1996

Cloning and characterization of the rat neurotensin receptor gene promoter.

The 5'-terminal region of the rat neurotensin receptor (NTR) gene was isolated and characterized. Genomic Southern analysis revealed that the NTR gene occurs as a single copy in the rat haploid genome. The upstream putative promoter region did not contain canonical TATA or CAAT boxes but has a consensus sequence for the transcription factor Sp1. This promoter is embedded in a large G + C-rich domain with characteristics of an CpG island. Transfection experiments using neurotensin receptor-luciferase fusion genes demonstrated that the 5'-flanking sequence functions as a strong promoter in the NG-108-15 cell. Deletion analysis suggested the presence of a core promoter (-470 to -662) that drives the minimal expression of the NTR gene.

Animals↗