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Figure 1 Protocol of studies to determine the time course of remote ischemic preconditioning (RIPC). Flow-mediated dilation (FMD) of the brachial artery was assessed before 20 min of arm ischemia (I) and at 20 min of reperfusion (R) (a). The effect of IR on brachial artery smooth muscle function was determined by measuring dilation in response to sublingual glyceryl trinitrate (GTN) (25 µg) administered before and after IR (b). Because pilot studies had shown that GTN prevented endothelial IR injury when administered immediately but not 24 h before, the control dilator response to GTN was determined 24 h before IR. The effect of RIPC of the contralateral arm on endothelial IR injury was determined by applying the RIPC stimulus immediately before IR (c). To determine the time course of protection by RIPC, the RIPC stimulus was applied 4 h (d), 24 h (e), and 48 h (f) before IR.
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