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Enzymatic desulfurization of dibenzothiophene by a cell-free system of Rhodococcus erythropolis D-1 总被引:1,自引:0,他引:1
Abstract Pseudomonas syringae cells were exposed to Cu2+ alone or in the precence of acetate, proline or cysteine, at concentrations that reduced free Cu2+ to 1/10 of the total copper. Ligand concentrations (designated as isoeffective) were determined experimentally using a Cu2+ -selective electrode and confirmed by computer calculations using published stability constants. Exposure of P. syringae cells to Cu2+ alone resulted in rapid and pronounced cell death, and binding of most of the copper in solution. The addition of acetate, proline or cysteine, a few minutes after Cu2+ treatment, resulted in a significant reduction in cell death, and in the amount of copper bound to the cells. For short exposures to Cu2+ , cysteine was more effective than acetate or proline, but after 60 min of treatment, similar results were observed with these ligands. The addition of ligands before Cu2+ resulted in even more reduced copper toxicity. The results showed that, at isoeffective concentrations, weak and moderate copper-ligands can effectively antagonize copper toxicity, and that this protective effect does not require previously equilibrated copper-ligand solutions and is not very dependent of the nature of the ligand. 相似文献
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Calcium mobilization and influx during the biphasic cytosolic calcium and secretory responses in agonist-stimulated pituitary gonadotrophs 总被引:3,自引:0,他引:3
Stimulation of enriched pituitary gonadotrophs by gonadotropin-releasing hormone (GnRH) elicits dose-dependent biphasic elevations of cytosolic calcium ([Ca2+]i) and luteinizing hormone (LH) release, with rapid initial peaks followed by sustained plateaus during continued exposure to the agonist. A potent GnRH-antagonist, [N-acetyl-D-p-Cl-Phe1,2,D-Trp3,D-Lys6,D-Ala10]GnRH, prevented the biphasic [Ca2+]i and LH responses when added before GnRH, and rapidly abolished both responses to GnRH when added during the plateau phase. In low Ca2+ medium the LH peak responses to GnRH were reduced and the subsequent sustained responses were almost completely abolished; reduction of extracellular Ca2+ during exposure to GnRH caused a prompt decline of LH release. The initial [Ca2+]i peak is derived largely from intracellular calcium mobilization with a partial contribution from calcium influx, while the sustained phase is dependent on the entry of extracellular Ca2+ through both L-type and dihydropyridine-insensitive channels. The presence of L-type voltage-sensitive calcium channels (VSCC) in pituitary gonadotrophs was indicated by the ability of elevated extracellular [K+] to stimulate calcium influx and LH release, and the sensitivity of these responses to dihydropyridine agonist and antagonist analogs. In cells pretreated with high [K+], the peak [Ca2+]i response to GnRH was enhanced but the subsequent plateau phase was markedly attenuated. This divergent effect of sustained membrane depolarization on the biphasic [Ca2+]i response suggests that calcium entry through VSCC initially potentiates agonist-induced mobilization of Ca2+ from intracellular storage sites. However, established Ca2+ entry through depolarization-activated VSCC cannot be further increased by agonist stimulation because both processes operate through the same channels, probably by changes in their activation-inactivation kinetics. Finally, the reciprocal potentiation by the dihydropyridine agonist, BK 8644, and GnRH of [Ca2+]i and LH responses confirms that both compounds act on the same type of channels, i.e., L-type VSCC, that participate in agonist-mediated calcium influx and gonadotropin secretion. 相似文献
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Yoshiaki Inayama Izumi Tomiyama Hitoshi Kitamura Yukio Nakatani Takaaki Ito Akinori Nozawa Yasuhiro Usuda Masayoshi Kanisawa 《Histochemistry and cell biology》1995,104(3):191-198
The purpose of this study was to investigate alkaline phosphatase (ALPase) reactivity in rabbit airway epithelial cells. Acetone-fixed, methyl benzoate and xylene-cleared (AMeX-treated) paraffin sections of trachea, bronchus, and lung tissue were stained by an azo dye coupling method for ALPase and examined by light microscopy. Electron histochemical staining was also performed in order to study the sensitivity and specificity of reactivity in each cell type. ALPase reactivity at the light microscopic level was observed exclusively in trachco-bronchial basal cells, and not in bronchiolar basal cells. By electron microscopy, ALPase reactivity was noted in 97.9% of basal cells in the trachea, 97.0% of basal cells in the bronchus, and 94.5% of basal cells and 15.4% of Clara cells in the bronchiole. This was also true for dispersed tracheal epithelial cells. Reactivity was rarely observed in ciliated cells, non-goblet-type secretory cells, and undetermined cells. The reactivity was heatlabile, levamisole-sensitive, and of a non-specific type. These findings indicate that basal cells of rabbit trachea and bonchus have fairly high specificity for ALPase of a non-specific isozyme (92.2% and 95.6%, respectively). Therefore, ALPase is considered to be a useful marker for these cells. 相似文献
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Evidence to show the presence of glucose-6-phosphate dehydrogenase,6-phospho-gluconate dehydrogenase, and NADP-dependent malicenzyme in proplastids of in vitro-cultured tobacco cells wasobtained. Amino acid synthesis from nitrite and 2-oxoglutaratein the proplastids was stimulated by addition of 20 mM glucose-6-phosphate.6-Phosphogluconate, malate, and isocitrate did not affect thesynthesis. Nitrite reduction and glutamate synthesis in theproplastids are assumed to be supplied with NADPH2 as the sourceof reducing power through the reactions catalyzed by glucose-6-phosphatedehyrdogenase and 6-phosphoglyconate dehydrogenase. (Received March 22, 1977; ) 相似文献
10.
Izumi Nakashima Fumihiko Nagase Akio Matsuura Takashi Yokochi Nobuo Kato 《Cellular immunology》1980,52(2):429-437
We demonstrated that each of various polyclonal lymphocyte activators (PLA) exhibits two types of adjuvant action to initiate the carrier-specific helper T-cell response to otherwise nonimmunogenic antigen. Type 1 action was characterized as that to initiate the T-cell response to subcutaneous injection of soluble bovine γ-globulin (BGG), and type 2 as that to initiate the response to intravenous injection of aggregated BGG. Each of various PLA showed these two types of adjuvant action in a dissociated fashion. The capsular polysaccharide of Klebsiella pneumoniae (CPS-K) showed both types of action to the highest degrees. Lipopolysaccharide of Escherichia coli exhibited type 2 action as markedly as CPS-K, but failed to show type 1 action. Concanavalin A showed definite type 1 action, but not type 2 action. Polyadenylic-uridylic acid showed definite type 2 action, but not type 1 action. Type 1 and type 2 actions of dextran sulfate were minimal. A hypothetical view is presented to consider that type 1 adjuvant action is directed to two mutually independent sites whereas type 2 action is directed to one site. 相似文献