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The protocol and consent were approved by the Institutional Review Boards at the University of Texas Southwestern Medical Center at Dallas and Texas Health Presbyterian Hospital Dallas. On arrival in the laboratory, ?90 Alectinib cost min prior to the onset of data collection, subjects swallowed an ingestible telemetry pill for the measurement of core temperature (HQ, Palmetto, FL, USA). With the exception of water ingested with this telemetry pill, subjects refrained from ingestion of fluids throughout the protocol. Each subject donned a water-perfused tube-lined suit (Med-Eng, Ottawa, Ontario, Canada) that covered their entire body except for the head, hands, left forearm and feet. The suit permitted the control of whole-body skin and internal temperatures by adjusting the temperature Osimertinib price of the water perfusing the suit. Weighted mean skin temperature was obtained from six thermocouples (Taylor et al. 1989) fixed to the skin under the water-perfused suit with porous adhesive tape. Arterial blood pressure was continuously measured non-invasively using photoplethysmography (Finometer Pro; FMS, Amsterdam, The Netherlands) and was also measured at the brachial artery by auscultation (Tango; Suntech Medical Instruments, Raleigh, NC, USA). Heart rate was obtained from an electrocardiogram (ECG; Agilent, Munich, Germany) that was connected to a cardiotachometer (1000 Hz sampling rate; CWE, Ardmore, PA, USA). Skin blood flow was measured at two sites on the dorsal portion of the left forearm, both of which were not exposed to the water-perfused suit. Each laser Doppler flow probe, which provides an integrated signal from GPX4 multiple emitting and receiving light sources (Periflux413; Perimed, North Royalton, OH, USA), was connected to a laser Doppler flowmeter (Periflux 5010; Perimed). Each probe was fitted inside a local heating device (Peritemp 4005; Perimed) capable of controlling skin temperature at that site. The local heater and laser Doppler probe assembly was placed on the skin over a fine-wire skin thermocouple (RET 4, type T thermocouple; Physitemp, Clifton, NJ, USA) such that local skin temperatures were measured at the site of skin blood flow measurement. Local heaters remained off during normothermic baseline data collection. In concert with the onset of heat stress, and throughout heat stress, local temperature at one site was elevated to approximate mean skin temperature under the water-perfused suit; achieving a level of ?38��C (defined as moderate). The heater for the adjacent probe assembly remained off throughout the heat stress (defined as mild), and thus temperature at that site changed secondary to reflex-induced cutaneous vasodilatation; an approach similar to prior evaluations (Johnson et al. 1973; Rowell et al. 1973; Tripathi & Nadel, 1986; Kellogg et al. 1990; Crandall et al. 1996, 2010; Shibasaki et al. 2006). Following instrumentation, subjects rested supine for 30 min, sealed at the waist within an LBNP device.