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Biomedical subjects

Y Oyama

Publications and source records attributed to Y Oyama.

At least 109 records · Page 6Linked to original sources

Flow cytometric analysis on cytotoxic action of amyloid beta protein fragment 25-35 on brain neurons dissociated from the rats.

Effects of amyloid beta protein fragment 25-35, A beta P(25-35), on membrane permeability and cell viability were examined in the brain neurons dissociated from the rats using a flow cytometer and two fluorescent dyes, fluo-3 to monitor intracellular Ca2+ concentration ([Ca2+]i) of neurons and ethidium which is impermeant to membranes of intact neurons to stain dead and dying neurons. A beta P(25-35) augmented fluo-3 fluorescence of some neurons at concentrations greater than 1 microM, indicating an increase in [Ca2+]i although other neurons (about 80% of total neurons) did not respond to A beta P(25-35) even at 10 microM. A beta P(25-35) at 1 microM or greater increased dose-dependently the number of ethidium-stained neurons, suggesting a dose-dependent increase in number of dead and dying neurons. Results suggest that A beta P(25-35) increases the membrane permeability of brain neurons, resulting in a destabilized intracellular homeostasis that leads to neuonal death.

Amyloid beta-Peptides↗

Methylmercury-induced augmentation of oxidative metabolism in cerebellar neurons dissociated from the rats: its dependence on intracellular Ca2+.

Effect of methylmercury chloride on oxidative metabolism of cerebellar neurons dissociated from the rats was examined using 2',7'-dichlorofluorescin (DCFH) which is oxidized by cellular hydrogen peroxide to be a fluorescent compound (DCF) and fluo-3, an indicator for intracellular Ca2+ concentration ([Ca2+]i). Methylmercury at 1 microM or less did not affect DCF fluorescence of cerebellar neurons. Further increase in concentration of methylmercury (up to 30 microM) induced changes in DCF fluorescence. Thus, DCF fluorescence was slightly attenuated during 5 min after applying methylmercury to the neurons, indicating a decrease in oxidation of DCFH. Thereafter, DCF fluorescence was time-dependently augmented in continued presence of methylmercury, indicating an increase in DCFH oxidation. Although methylmercury-induced augmentation of DCF fluorescence was greatly suppressed under external Ca(2+)-free condition, it was not the case for methylmercury-induced attenuation of DCF fluorescence. Methylmercury at 3 microM or more dose-dependently increased the [Ca2+]i. Results suggest that methylmercury increases intracellular Ca2+ in cerebellar neurons, resulting in an increase in formation of reactive oxygen species that may contribute to cell injury.

Aniline Compounds↗

Effect of N,N-diethyldithiocarbamate on ionomycin-induced increase in oxidation of cellular 2',7'-dichlorofluorescin in dissociated cerebellar neurons.

Effect of N,N-diethyldithiocarbamate (DDC), an inhibitor for cytosolic superoxide dismutase, on an ionomycin-induced increase in oxidative metabolism was examined in cerebellar neurons dissociated from the rats, using a flow cytometer and 2',7'-dichlorofluorescin diacetate and fluo-3-AM, fluorescent dyes for intracellular hydrogen peroxide and Ca2+, respectively. DDC reduced the ionomycin-induced augmentation of 2',7'-dichlorofluorescin fluorescence in a dose-dependent manner. DDC did not affect cellular content of 2',7'-dichlorofluorescin and ionomycin-induced increase in intracellular Ca2+ concentration. Results indicate that ionomycin increases the formation of superoxide anion in brain neuron.

Aniline Compounds↗

Quantitative autoradiographic distribution of glutamate receptors in the cervical segment of the spinal cord of the wobbler mouse.

The reduction of glutamate content has been observed in the spinal cord of the wobbler mouse, a purported model of amyotrophic lateral sclerosis (ALS). To elucidate glutamate receptors in the wobbler spinal cord, we measured densities of N-methyl-D-aspartate (NMDA), kainate, alpha-amino-3-hydroxy-5-methyl-4-isoxazole proprionic acid (AMPA) and metabotropic glutamate (mGlu) binding sites using in vitro autoradiography. In wobbler mice, NMDA, kainate, and AMPA binding sites were increased in the dorsal horn and kainate binding sites were also increased in the intermediate zone. However, mGlu binding was unchanged. These results disagree with those observed in ALS spinal cords, in which NMDA and kinate binding sites are decreased. The wobbler mouse may have the glutamate dysfunction, but in a different way from ALS.

Animals↗

Effect of tri-n-butyltin on intracellular Ca2+ concentration of mouse thymocytes under Ca(2+)-free condition.

Effect of tri-n-butyltin at concentrations ranging from 100 nM to 1 microM on the intracellular Ca2+ concentration of mouse thymocytes was examined under Ca(2+)-free conditions in comparison with those of 50 nM A23187, 100 nM thapsigargin and 10 microM cyclopiazonic acid, using the fluorescent dye for intracellular Ca2+, fluo-3. Tri-n-butyltin persistently increased the intensity of fluo-3 fluorescence while A23187, thapsigargin and cyclopiazonic acid produced a transient augmentation of the fluorescence. Pretreatment with A23187 greatly decreased the fluorescence responses induced by 1 microM tri-n-butyltin. However, the effect of thapsigargin and cyclopiazonic acid on the tri-n-butyltin-induced response was much weaker than that of A23187. In the presence of tri-n-butyltin, the transient response produced by A23187 was greatly prolonged. Results may suggest that tri-n-butyltin increases the membrane Ca2+ permeability of the intracellular organelles (cellular calcium stores) and decreases the Ca2+ pump activity of thymocyte membrane, resulting in a sustained increase in the intracellular Ca2+ concentration under Ca(2+)-free concentration.

Aniline Compounds↗

Characterization of 2',7'-dichlorofluorescin fluorescence in dissociated mammalian brain neurons: estimation on intracellular content of hydrogen peroxide.

The fluorescence of 2',7'-dichlorofluorescin (DCF) was measured in acutely dissociated rat cerebellar neurons as a mean of estimating the formation of reactive oxygen species (ROS). N,N-Diethyldithiocarbamate, an inhibitor for superoxide dismutase, reduced the intensity of DCF fluorescence in a dose-dependent fashion at concentrations of 30 nM to up to 10 microM. N-Ethylmaleimide, an inhibitor for glutathione peroxidase, augmented the DCF fluorescence in a dose-dependent manner at concentration of 10 microM to 1 mM while 3-amino-1,2,4-triazole, an inhibitor for catalase, did not change the fluorescence intensity even at concentrations as high as 1 mM. Hydrogen peroxide, applied externally at concentrations between 3 microM and 3 mM, augmented the fluorescence in a dose-dependent fashion. These results suggest the possibility that the DCF fluorescence may be useful in estimating the intracellular content of hydrogen peroxide of mammalian brain neurons.

Animals↗

Myricetin and quercetin, the flavonoid constituents of Ginkgo biloba extract, greatly reduce oxidative metabolism in both resting and Ca(2+)-loaded brain neurons.

The antioxidant action of myricetin and quercetin, the flavonoid constituents of the extract of Ginkgo biloba (EGb), on oxidative metabolism of brain neurons dissociated from the rats was examined using 2',7'-dichlorofluorescin (DCFH) which is retained within the neuron and then is oxidized by cellular hydrogen peroxide to be highly fluorescent. Incubation with myricetin or quercetin reduced the oxidation of DCFH in resting brain neurons, more profoundly than EGb. Myricetin decreased the oxidative metabolism at concentrations of 3 nM or more. It was 10 nM or more for the case of quercetin. Incubation with each flavonoid constituent also reduced the Ca(2+)-induced increase in the oxidative metabolism without affecting the cellular content of DCFH or the intracellular concentrations of Ca2+. Such an antioxidant action of myricetin or quercetin may be responsible for a part of the beneficial effects of EGb on brain neurons subject to ischemia.

Animals↗

[Thallium-201 single photon emission computed tomography in the evaluation of therapeutic response for brain tumors].

To evaluate the effectiveness of radiation and chemotherapy of malignant brain tumors, 201TlCl-SPECT (Tl-SPECT) was compared with CT and MRI. The subjects were 14 patients with malignant brain tumors; 12 malignant astrocytomas, 1 medulloblastoma and 1 malignant lymphoma. Tl-SPECT was performed before and immediately after the therapy. On the Tl-SPECT, count ratio (Tl-index) of tumor to normal brain tissue was measured. To evaluate the size of tumor, CT or MRI was performed before and immediately after the therapy, and 1-2 months after the therapy. The change of Tl-index immediately after therapy was compared with the change of the size of tumor. Though the change of Tl-index did not correlate with the change of tumoral size immediately after therapy, it correlated with the change of tumoral size 1-2 months after the therapy, i.e., it was thought that Tl-index reflected the change of tumoral size 1-2 months after the therapy. Tl-SPECT will be feasible for the evaluation of the effectiveness of the therapy for malignant brain tumors, because it reflects the effectiveness of the therapy earlier than CT or MRI.

Adult↗

Effect of KB-2796, a new diphenylpiperazine Ca2+ antagonist, on voltage-dependent Ca2+ currents and oxidative metabolism in dissociated mammalian CNS neurons.

The effects of KB-2796, 1-[bis(4-fluorophenyl)methyl]-4-(2,3,4- trimethoxybenzyl)piperazine-2HCl, on the low- and high-voltage activated Ca2+ currents (LVA and HVA ICa, respectively) and on oxidative metabolism were studied in neurons freshly dissociated from rat brain. KB-2796 reduced the peak amplitude of LVA ICa in a concentration-dependent manner with a threshold concentration of 10(-7) M when the LVA ICa was elicited every 30 s in the external solution with 10 mM Ca2+. The concentration for half-maximum inhibition (IC50) was 1.9 x 10(-6) M. At 10(-5) M or more of KB-2796, a complete suppression of the LVA ICa was observed in the majority of neurons tested. There was no apparent effect on the current-voltage (I-V) relationship and the current kinetics. KB-2796 delayed the reactivation and enhanced the inactivation of the Ca2+ channel for LVA ICa voltage- and time-dependently, suggesting that KB-2796 preferentially binds to the inactivated Ca2+ channel. KB-2796 at a concentration of 3.0 x 10(-6) M also decreased the peak amplitude of the HVA ICa without shifting the I-V relationship. In addition, KB-2796 reduced the oxidative metabolism (the formation of reactive oxygen species) of the neuron in a concentration-dependent manner with a threshold concentration of 3 x 10(-6) M. It is suggested that the inhibitory action of KB-2796 on the neuronal Ca2+ influx and the oxidative metabolism, in combination with a cerebral vasodilatory action, may reduce ischemic brain damage.

Animals↗

Effect of tri-n-butyltin on intracellular Ca2+ concentration of rat cerebellar neurons.

The effect of tri-n-butyltin on the intracellular Ca2+ concentration ([Ca2+]i) of cerebellar neurons dissociated from rats was examined using fluo-3 and a flow cytometer. Tri-n-butyltin at 100 nM or more (up to 1 microM) increased the [Ca2+]i in a dose-dependent manner. The effect of tri-n-butyltin on the [Ca2+]i was greatly reduced under external Ca(2+)-free ([Ca2+]o-free) conditions, suggesting its dependence on the presence of [Ca2+]o. Lower trialkyltins, such as triethyltin and trimethyltin at 1 microM, exerted little or no action on the [Ca2+]i. Therefore, the cytotoxic action of tri-n-butyltin may be different from those of lower trialkyltins.

Animals↗

Fluorescent estimation on the effect of Ca2+ antagonists on the oxidative metabolism in dissociated mammalian brain neurons.

Effect of organic Ca2+ antagonists on the oxidative metabolism or the formation of reactive oxygen species was estimated on dissociated mammalian neurons using a flow cytometer and 2',7'-dichlorofluorescin which is oxidized to be fluorescent by cellular oxidants. Of the organic Ca2+ antagonists used, flunarizine and nifedipine decreased the intensity of fluorescence at 1 microM or more while it was not the case for verapamil and diltiazem, suggesting one of the favourable actions of flunarizine and nifedipine on ischemic brain damage.

Animals↗

[Discarding medical images and its evaluation].

Following a survey of our filing system, the period of storage of medical images was generally limited to ten years, a system which was put into execution in 1990. Film jackets older than ten years are discarded every year, except for those registered for permanent storage. Fourteen thousand jackets (6.5% of total) have been registered for permanent storage. A follow-up survey has been performed to evaluate this system. The survey focused mainly on the frequency of re-issue of film jackets, and the rate of access to jackets. Discarded jackets now number about 50,000, a reduction of 20%. A total of 405 jackets were re-issued in a year due to the re-examination of patients whose jackets had been discarded. The annual number of re-called jackets from the repository outside our hospital was about 2,200 (about 500 of them were for research work), which corresponded to 1.5% of the patients re-examined in the radiology department. The new filing system has been well integrated into the hospital daily routine. Information cost was calculated as a function of access rate to the stored information, and the concept of "cost-of-information" was proposed as a measure of cost analysis of an information system.

Costs and Cost Analysis↗

Ca(2+)-induced increase in oxidative metabolism of dissociated mammalian brain neurons: effect of extract of ginkgo biloba leaves.

Effect of an extract of Ginkgo biloba leaves (EGb) on oxidative metabolism was studied using rat brain neurons and 2',7'-dichlorofluorescin fluorescence. Ionomycin (100 nM to 1 microM), a Ca(2+)-ionophore, dose-dependently augmented the 2',7'-dichlorofluorescin fluorescence in the presence of external Ca2+, but not under the external Ca(2+)-free condition. Preincubation of neurons with EGb (3 micrograms/ml) greatly reduced the ionomycin-induced increase in 2',7'-dichlorofluorescin fluorescence. Results suggest that EGb may reduce the Ca(2+)-induced increase in the oxidative metabolism of brain neurons.

Animals↗

Computed tomography and magnetic resonance findings of meningeal syndrome in a leukemic cat.

A 3-year-old Japanese domestic cat with a diagnosis of lymphocytic leukemia showed severe generalized seizures in the course of chemotherapy after leukemic condition was improved clinically. Computed tomography (CT) and magnetic resonance (MR) imaging of the brain were carried out. Both contrast procedures disclosed enhancements at the falx cerebri and the margin of cerebral cortex. Among these procedures contrast MR imaging demonstrated the lesion most clearly. Cytological examination of cerebrospinal fluid obtained by spinal puncture showed the infiltration of malignant cells and the diagnosis of meningeal syndrome associated with lymphocytic leukemia was defined. Intrathecal administration of cytosine arabinoside partially improved the neurologic dysfunction. Autopsy and histopathological examination confirmed the infiltration of leukemic cells in the areas of meningeal lesion demonstrated with contrast CT and MR imaging. Thus these imaging techniques, especially contrast MR imaging, are useful tools for rapid and precise diagnosis of meningeal syndrome.

Animals↗

Modification of voltage-dependent Na+ current by triphenyltin, an environmental pollutant, in isolated mammalian brain neurons.

In order to reveal the profile of neurotoxicity of triphenyltin on the mammalian central nervous system (CNS) through the modification of a voltage-dependent Na+ channel, the effects of triphenyltin on the kinetics of voltage-dependent Na+ current (INa) were examined in acutely dissociated pyramidal neurons of rat hippocampus. Triphenyltin at the concentration of 1.10(-6) M decreased both the time to peak and the half-decay time of the INa without affecting the current-voltage relationship. Triphenyltin moved the steady-state inactivation curve to a depolarizing direction. In the presence of triphenyltin, the reactivation of Na+ channel inactivated during the depolarization occurred quicker than that of the control at the pulse intervals of less than 10 ms. It can be suggested that triphenyltin potentially increases the cell excitability in mammalian CNS through the modification of the INa.

Animals↗