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The tuber necrotic strain of Potato virus Y (PVYNTN) causes widespread disease and has severe negative effects on the growth and yields of plants, especially those of the Solanaceae family. The consequences of residual toxicity and non-biodegradation of synthetic chemicals and pollution of the environment has led to investigations into new non-toxic and biological treatments to control plant viral diseases. Ethanolic extracts of Bowiea volubilis (bulbs), Cotyledon orbiculata (leaves), Gomphocarpus fruticosus (leaves), Merwilla plumbea (dry and fresh bulbs), Nerium oleander (leaves), and the fruits and leaves of Strophanthus speciosus, were evaluated against PVYNTN in vivo and in vitro. At a concentration of 20 mg · ml−1, ethanolic extracts of Strophanthus speciosus (leaves) and fruits (50 mg · ml−1) significantly reduced the expression of PVYNTN symptoms on tobacco plants in vitro without affecting the normal growth and development of the plant. Similarly, at 50 mg · ml−1, N. oleander, C. orbiculata and B. volubilis (fresh bulbs) and S. speciousus leaves at 20 mg · ml−1 extracts showed significant differences in PVYNTN symptoms in the in vivo experiment. Strophanthus speciosus leaf and fruit extracts showed significant inhibition in the in vitro and in vivo assays and demonstrated that S. speciosus has potential to be used as an antiphytoviral treatment.
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.
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