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Tormo Nuria Giménez Estela Martínez-Navarro María Albert Eliseo Navalpotro David Torres Ignacio Gimeno Concepción Navarro David 《European journal of clinical microbiology & infectious diseases》2022,41(4):657-662
European Journal of Clinical Microbiology & Infectious Diseases - We compared the performance of an in-house-developed flow cytometry assay for intracellular cytokine staining (FC-ICS) and a... 相似文献
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The glucose-clamp technique was used to assess the effect on insulin activity of mixing clinically relevant doses of short- and longer-acting insulins in insulin-treated diabetic subjects. Mixtures of porcine regular and lente insulins resulted in a reduction in activity of the mixture compared with separate injection that was more apparent with premixing for 5 min before injecting than with premixing for 2 min. These findings were not explained by differences in volume of the injected insulin preparations. Mixing of porcine regular and bovine ultralente insulins for 2 min before injecting resulted in a reduced activity compared with separate injection. No difference in activity was obvious for porcine regular and NPH insulins given separately or premixed for 5 min. A reduction of activity during the 1st h after injection was observed when separate injection of one brand of these insulins (Velosulin or Insulatard) was compared with the manufactured mixture (Mixtard). These results indicate that premixing regular and NPH insulins in a 1-to-2 ratio does not alter the biological activity compared with separate administration of the insulins. However, the mixing of regular and insulin-zinc suspensions results in a loss of insulin activity, the magnitude of which depends on the time between mixing and injecting the insulin. The clinical significance of the effect of mixing on the efficacy of subcutaneous insulin therapy should be considered in the context that this is only one of many factors that may affect the activity of subcutaneously injected insulin. 相似文献
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Lucía Núñez Ruth A. Valero Laura Senovilla Sara Sanz-Blasco Javier García-Sancho Carlos Villalobos 《The Journal of physiology》2006,571(1):57-73
Store-operated Ca2+ entry (SOCE) is a ubiquitous Ca2+ influx pathway involved in control of multiple cellular and physiological processes including cell proliferation. Recent evidence has shown that SOCE depends critically on mitochondrial sinking of entering Ca2+ to avoid Ca2+ -dependent inactivation. Thus, a role of mitochondria in control of cell proliferation could be anticipated. We show here that activation of SOCE induces cytosolic high [Ca2+ ] domains that are large enough to be sensed and avidly taken up by a pool of nearby mitochondria. Prevention of mitochondrial clearance of the entering Ca2+ inhibited both SOCE and cell proliferation in several cell types including Jurkat and human colon cancer cells. In addition, we find that therapeutic concentrations of salicylate, the major metabolite of aspirin, depolarize partially mitochondria and inhibit mitochondrial Ca2+ uptake, as revealed by mitochondrial Ca2+ measurements with targeted aequorins. This salicylate-induced inhibition of mitochondrial Ca2+ sinking prevented SOCE and impaired cell growth of Jurkat and human colon cancer cells. Finally, direct blockade of SOCE by the pyrazole derivative BTP-2 was sufficient to arrest cell growth. Taken together, our results reveal that cell proliferation depends critically on mitochondrial Ca2+ uptake and suggest that inhibition of tumour cell proliferation by salicylate may be due to interference with mitochondrial Ca2+ uptake, which is essential for sustaining SOCE. This novel mechanism may contribute to explaining the reported anti-proliferative and anti-tumoral actions of aspirin and dietary salicylates. 相似文献
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Carlos Villalobos Javier García-Sancho 《Pflügers Archiv : European journal of physiology》1996,431(3):371-378
Caffeine, a well known facilitator of Ca2+-induced Ca2+ release, induced oscillations of cytosolic free Ca2+ ([Ca2+]i) in GH3 pituitary cells. These oscillations were dependent on the presence of extracellular Ca2+ and blocked by dihydropyridines, suggesting that they are due to Ca2+ entry through L-type Ca2+ channels, rather than to Ca2+ release from the intracellular Ca2+ stores. Emptying the stores by treatment with ionomycin or thapsigargin did not prevent the caffeine-induced [Ca2+]i oscillations. Treatment with caffeine occluded phase 2 ([Ca2+]i oscillations) of the action of thyrotropin-releasing hormone (TRH) without modifying phase 1 (Ca2+ release from the intracellular stores). Caffeine also inhibited the [Ca2+]i increase induced by depolarization with high-K+ solutions (56% at 20 mM), suggesting direct inhibition of the Ca2+ entry through voltage-gated Ca2+ channels. We propose that the [Ca2+]i increase induced by caffeine in GH3 cells takes place by a mechanism similar to that of TRH, i.e. membrane depolarization that increases the firing frequency of action potentials. The increase of the electrical activity overcomes the direct inhibitory effect on voltage-gated Ca2+ channels with the result of increased Ca2+ entry and a rise in [Ca2+]i. Consideration of this action cautions interpretation of previous experiments in which caffeine was assumed to increase [Ca2+]i only by facilitating the release of Ca2+ from intracellular Ca2+ stores. 相似文献