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Content available In vitro cultures of animal and human cells
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EN
Cytological studies have been performed on Allium sibiricum L. callus culture. Interesting cytological changes observed during callus growth included polyploidy, aneuploidy and structural aberrations of chromosomes (ring-chromosomes, deletions, formation of dicentric and tricentric chromosomes). The rate of these changes increased with the time of culture.
EN
In circulation, platelets may come into contact with both exogenous (cardiac glycoside treatment) and endogenously produced inhibitors of Na+/K+-ATPase. We examined whether blocking of platelet Na+/K+-ATPase by ouabain results in generation of procoagulant activity. It was shown that an in vitro treatment of platelets with ouabain (20-200 µM for 20 to 60 min) is associated with an intracellular accumulation of sodium ([Na+]i), generation of a weak calcium signal, and expression of procoagulant activity. The ouabain-induced procoagulant response was dose- and time-related, less pronounced than that evoked by collagen and similar to that produced by gramicidin, not affected by EDTA or aspirin, and strongly reduced in the absence of extracellular Na+ or by hyperosmolality. Flow cytometry studies revealed that ouabain treatment results in a unimodal left shift in the forward and side scatter of the entire platelet population indicating morphological changes of the plasma membrane. The shift was dose related, weaker than that evoked by collagen and similar to that produced by gramicidin. Ouabain-treated platelets express phosphatidylserine (PS). The ouabain-evoked PS expression was dose- and time-dependent, weaker than that produced by collagen and similar to that evoked by gramicidin. Electronic cell sizing measurements showed a dose-dependent increase in mean platelet volume upon treatment with ouabain. Hypoosmotically-evoked platelet swelling resulted in the appearance of procoagulant activity. Thromboelastography measurements indicate that, in whole blood, nanomolar (50-1000 nM, 15 min) concentrations of ouabain significantly accelerate the rate of clot formation initiated by contact and high extracellular concentration of calcium. We conclude that inefficiently operating platelet Na+/K+-ATPase results in a rise in [Na+]i. An increase in [Na+]i and the swelling associated with it may produce PS exposure and a rise in membrane curvature leading to the generation of a procoagulant activity.
EN
The eutrophication of the Sulejów reservoir dam in Poland is connected with the problem of toxicity of cyanobacterial blooming (blue-green algal blooming). The main species responsible for hepatotoxic ”algal bloom” formation is Microcystis aeruginosa. The aim of this study is to evaluate the influence of the toxic cyanobacterial extract containing microcystins on the growth and morphology of a water plant (Spirodela oligorrhiza) and animal cells (rat hepatocytes). A higher concentration of cyanobacterial extract (MC-LR = 343 µg/dm3) reduced the number of fronds by about 50% in comparison with the control. The extract affected the reduction mass of fronds and the concentration of chlorophyll. The activity of the constitutive acid phosphatase decreased. The first morphological changes in rat hepatocytes typical of apoptosis were observed after 30 minutes of incubation with the cyanobacterial extract. The hepatocytes underwent cell membrane blebbing (MC-LR = 100 µg/dm3). The next 30 minutes of incubation caused an increase in the percentage of deforming cells of more than 50% (MC-LR = 100 µg/dm3). High chromatin condensation and apoptotic bodes were observed in 90% of cells after 120 minutes (MC-LR . 500 µg/dm3). The results of studies confirm the high toxic and cytotoxic effect of blue-green algal blooming from Sulejów reservoir on both plants and animals.
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