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Biomedical subjects

C Yamamoto

Publications and source records attributed to C Yamamoto.

At least 163 records · Page 9Linked to original sources
PubMed1993

Characterization of tumor necrosis factor alpha-induced alteration of glycosaminoglycans in cultured cells: comparison among vascular smooth-muscle cells, vascular endothelial cells, Chang liver cells and LLC-PK1 cells.

We investigated the alteration of glycosaminoglycans (GAGs) induced by recombinant human tumor necrosis factor alpha (rhTNF alpha) using confluent cultures of bovine aortic smooth-muscle cells, bovine aortic endothelial cells, Chang liver cells and porcine kidney LLC-PK1 cells. It was found that the incorporation of both [35S]sulfate and [3H]glucosamine into GAGs in the trypsinate fraction of the cell layer was significantly decreased by rhTNF alpha in vascular smooth-muscle cells and vascular endothelial cells; the incorporation of [35S]sulfate was increased but that of [3H]glucosamine was unchanged in Chang liver cells; the incorporation of both [35S]sulfate and [3H]glucosamine was increased by rhTNF alpha in LLC-PK1 cells. In the conditioned medium, the incorporation of both [35S]sulfate and [3H]glucosamine was not greatly changed by rhTNF alpha in all tested cell types. Characterization of GAGs revealed that each cell type uniquely altered its GAGs after rhTNF alpha treatment; the cytokine-induced alteration of each GAG component was not necessarily the same among different cell types. It was therefore concluded that rhTNF alpha-induced alteration of GAGs is dependent upon cell type.

Animals↗
PubMed1993

De-acclimatization process of rats acclimatized to high altitude hypoxia.

The de-acclimatization process in rats acclimatized to high altitude hypoxia was analyzed according to the changes in the activities of hepatic enzymes. The activities of tryptophan 2,3-dioxygenase and tyrosine aminotransferase in the liver of non-acclimatized rats were increased, whereas those of acclimatized rats were decreased after exposure to a simulated altitude of 8,000 m. When the acclimatized rats were kept under normoxic conditions for 8 days and over, the suppression of enzyme induction was nullified, and both enzymes were induced by exposure to the simulated 8,000 m altitude. De-acclimatization of rats acclimatized to hypoxia can be completed in 8-11 days.

Acclimatization↗
PubMed1993

Urinary excretion of stress hormones of rats in tail-suspension.

Tail-suspension of rats has been employed for the study of disuse atrophy of hindlimb muscles and bones. From our study, it is suggested that stress exacerbates muscle and bone atrophy; however, the determination of stress hormones has been inappropriately performed. For assessing the effect of tail suspension on stress hormone secretion, suspension was performed for 7 days in a metabolic cage. Urinary excretions of corticosterone, catecholamines and antidiuretic hormone were determined during the 7-day suspension. Urine volume was unchanged during the suspension period, whereas water consumption was decreased during suspension. Urinary excretion of corticosterone and catecholamines increased significantly during the initial 3 days of suspension. The excretion of antidiuretic hormone increased throughout the suspension period. Our results demonstrated that the determination of urinary excretion of these hormones is useful for evaluating the stress reaction.

Adrenal Glands↗
PubMed1993

Method of identifying inhibitors of oncogenic transformation: selective inhibition of cell growth in serum-free medium.

We developed a new method for evaluating inhibitors of oncogenic signal transduction pathways based on different growth abilities between normal and transformed cells in a defined serum-free medium. The growth rates of src, abl or ras oncogene-transformed cells, activated raf proto-oncogene transformed cells, and normal NIH-3T3 cells were 60-90%, 20-30% and 10% in a serum-free medium, respectively, compared to the growth rates in a serum-containing medium. An addition of a growth factor (PDGF, FGF or TGF-beta) stimulated the growth of normal NIH3T3 cells by 40-80% in a serum-free medium. Herbimycin A, a specific cytoplasmic protein tyrosine kinase inhibitor, selectively inhibited the growth of src or abl transformed cells in the serum-free medium resulting in about 10-fold or fivefold lower IC50 than those in the serum-containing medium. The antibiotic did not show such an effect on ras transformed cells, and the treatment of src transformed cells with other protein kinase inhibitors or cytotoxic drugs showed little IC50 shifts between the two media. Thus, this method of comparing growth inhibition in the serum-free and the serum-containing media may be useful in evaluating specific inhibitors of signaling pathways mediated by growth factors and certain oncogene products.

3T3 Cells↗
PubMed1992

Possible mechanism for zinc protection against cadmium cytotoxicity in cultured vascular endothelial cells.

We investigated the effect of zinc on cadmium cytotoxicity in vascular endothelial cells in a culture system. The cytotoxicity was evaluated by [3H]adenine release assay. Cadmium at 2 microM concentration and above significantly increased the [3H]adenine release, but zinc at 80 microM and below did not induce such a response after a 24-h incubation. Metallothionein was induced by cadmium at 0.1 microM and above but not by zinc at 300 microM and below; however, zinc at 10 microM and above significantly decreased cadmium-(2 and 5 microM) induced cytotoxicity. Zinc protection against cadmium cytotoxicity was also observed in the presence of 1 microM cycloheximide. Zinc caused significantly less accumulation of cadmium in the cell layer accompanied with a significant accumulation of zinc. The distribution (%) of cadmium in the particulate fraction of the cells was significantly decreased by zinc. In contrast, cadmium in the cytosol fraction was increased in the cells treated with both cadmium and zinc. Gel filtration chromatography of the cytosol showed that cadmium was capable of being bound to high-molecular-weight proteins and metallothionein. The metallothionein-bound cadmium was not increased by zinc; however, the relative distribution of cadmium in the high-molecular-weight fraction in the cytosol was decreased in cadmium plus zinc-treated cells. From these results, it was suggested that the mechanism by which zinc protects endothelial cells from cadmium cytotoxicity was decreased accumulation of cadmium in the particulate fraction and in the high-molecular-weight fraction in the cytosol of the cells. This alteration is postulated to be caused by both zinc-induced decrease in the intracellular cadmium accumulation and the sequestration of cadmium by cadmium-induced metallothionein.

Adenine↗
PubMed1992

Quantal analysis of suppressing action of baclofen on mossy fiber synapses in guinea pig hippocampus.

Baclofen, a selective agonist at the gamma-aminobutyric acidB (GABAB) receptor, has been considered to reduce the release of transmitter from nerve terminals by acting on presynaptic GABAB receptors, in addition to its postsynaptic action. The purpose of this study has been to re-examine quantitatively the action of baclofen in the hippocampus by a rigorous quantal analysis. (+/-)-Baclofen suppressed field potentials and intracellularly-recorded synaptic potentials induced in the subfield CA3 by mossy fiber stimulation in thin transverse sections of the guinea pig hippocampus. The amplitude distribution of excitatory postsynaptic potentials (EPSPs) induced monosynaptically by a granule cell could be described by the Pascal statistics. Suppression of the unitary EPSPs by baclofen (1 and 5 microM) was accompanied by decreases both in mean quantal content (m) and by mean quantal amplitude (q). The reduction in q was smaller than expected from a decrease in the input resistance of the postsynaptic neuron. It was suggested that the presynaptic and postsynaptic actions of baclofen contribute almost equally to suppression of the transmission at 1 microM, whereas the presynaptic action predominates at 5 microM.

Animals↗
PubMed1992

Enhancement of postsynaptic responsiveness during long-term potentiation of mossy fiber synapses in guinea pig hippocampus.

The primary site responsible for the long-lasting enhancement of synaptic transmission during long-term potentiation (LTP) was examined by quantal analysis of excitatory postsynaptic potentials in thin sections of the guinea pig hippocampus. With induction of LTP in mossy fiber synapses, estimated values of quantal amplitude (q) and Pascal parameters p and r were increased significantly. No increases in quantal content (m) were detected. The magnitude of increases in q was almost equal to that of LTP. These results indicate that LTP in mossy fiber synapses results from increases in responsiveness of postsynaptic neurons.

Afferent Pathways↗
PubMed1992

Plasmin stimulates the release of dermatan sulfate from vascular smooth muscle cells in culture.

We investigated the effect of plasmin on the release of sulfated glycosaminoglycans (GAG) from cultured vascular smooth muscle cells. Confluent cultures of vascular smooth muscle cells from bovine aorta were labelled with [35S]sulfate and incubated at 37 degrees C for principally 60 min in a serum-free medium in the presence of plasmin. Plasmin at 10 mU/ml (approximately 2.7 micrograms/ml) and above significantly increased the release of [35S]sulfate-labeled GAG (35S-GAG) from the cell layer after a 60 min incubation. A time course study showed that plasmin at 10 mU/ml significantly increased the 35S-GAG release after 20 min and longer. However, plasminogen at 100 mU/ml and below did not cause a significant change of the 35S-GAG release after a 90 min incubation. A characterization of 35S-GAG revealed that plasmin increased both dermatan sulfate and the other 35S-GAG in the medium. Plasmin at 100 mU/ml enhanced the cell detachment significantly but only slightly. From these results, it was suggested that a endogenous thrombin inhibitor heparin cofactor II may be activated in the liquid phase by dermatan sulfate released from plasmin-stimulated vascular smooth muscle cells when the vascular is disrupted and plasma is exposed to extravessel.

Animals↗
PubMed1992

Identification of an ATP-sensitive K+ channel in rat cultured cortical neurons.

To determine whether membranes of mammalian central neurons contain an ATP-sensitive K+ (KATP) channel similar to that present in pancreatic beta cells, the patch-clamp technique was applied to cultured neurons prepared from the neonatal rat cerebral cortex and hippocampus. In whole-cell experiments with hippocampal neurons, extracellular application of 0.5 mM diazoxide (a KATP channel activator) elicited a hyperpolarization concomitant with an increase in membrane conductance, whereas application of 0.5 mM tolbutamide (a KATP channel blocker) induced a depolarization with a decrease in conductance. Similar results were obtained with cortical neurons. In outside-out patch experiments with cortical neurons, a K+ channel sensitive to these drugs was found. The channel was completely blocked by 0.5 mM tolbutamide and activated by 0.5 mM diazoxide. The single-channel conductance was 65 pS under symmetrical 145 mM K+ conditions and 24 pS in a physiological K+ gradient. In inside-out patch experiments, this channel was demonstrated to be inhibited by an application of 0.2-1 mM ATP to the cytoplasmic surface of the patch membrane. These results indicate that the membranes of rat cortical neurons contain a KATP channel that is quite similar to that found in pancreatic beta cells. It is also suggested that the same or a similar K+ channel may exist in membranes of hippocampal neurons.

Adenosine Triphosphate↗
PubMed1992

An empirical test for the reliability of quantal analysis based on Pascal statistics.

We have previously shown that amplitude distributions of excitatory postsynaptic potentials (EPSPs) can be better described by Pascal distribution when the mean quantal content (m) is not stationary but fluctuating according to gamma distribution. We have developed the procedure of estimating quantal parameters by the method of maximum likelihood. In this study, we examined empirically the reliability of this quantal parameter estimation procedure by using Monte Carlo simulations. The reliability was evaluated by absolute values of error (magnitude of error) of estimated parameters relative to the known 'true' parameters. The mean values of relative magnitude of error were relatively small unless the probability of failures was too large (greater than 0.7) or too small (less than 0.1). The values of relative magnitude of error became smaller in association with increases in the sample size. When the probability of failure was between 0.1 and 0.7, the sample size was 1000, coefficient of variation of quantal size was 0.45, the values of relative magnitude of error of estimated parameters were below 0.1. These results mean that this procedure gives relatively reliable estimates of quantal parameters with the limitation that the probability of failure is neither too large (greater than 0.7) nor too small (less than 0.1); it is preferable that the sample size is as large as 1000.

Evoked Potentials↗
PubMed1992

Effect of cadmium on the monolayer maintenance of vascular endothelial cells in culture.

We investigated the effect of cadmium on the maintenance of the monolayer of vascular endothelial cells to clarify a possible involvement of endothelium injury in cadmium-induced vascular disorders. Endothelial cells from bovine aorta were cultured with cadmium chloride (0.5, 1.0, 2.0 or 5.0 microM) for 24 or 72 h. A histological observation revealed that a de-endothelialized space was formed in the monolayer by cadmium treatment; the number of endothelial cells was significantly decreased by the metal. Cadmium significantly decreased the number of growing endothelial cells with a significant decrease in the [3H]thymidine incorporation, suggesting that cadmium inhibited the cell proliferation. On the other hand, cadmium significantly increased the detachment of [3H]thymidine-labeled endothelial cells from the monolayer with a parallel increase in the lactate dehydrogenase activity in the medium. From these results, it was suggested that cadmium impairs the endothelial cell monolayer; the de-endothelialized space was formed by both an increase in the cell detachment by cadmium cytotoxicity and a retardation of the repair of the space which was due to an inhibition of the cell proliferation caused by the metal.

Animals↗