Nitric oxide synthase (NOS) contributes to sweating and cutaneous vasodilation during exercise in younger adults. We hypothesized that endothelial NOS (eNOS) and neuronal NOS (nNOS) mediate NOS-dependent sweating, whereas eNOS induces NOS-dependent cutaneous vasodilation in younger adults exercising in the heat. Further, aging may upregulate inducible NOS (iNOS), which may attenuate sweating and cutaneous vasodilator responses. We hypothesized that iNOS inhibition would augment sweating and cutaneous vasodilation in exercising older adults. Physically active younger (n = 12, 23 ± 4 yr) and older (n = 12, 60 ± 6 yr) adults performed two 30-min bouts of cycling at a fixed rate of metabolic heat production (400 W) in the heat (35°C). Sweat rate and cutaneous vascular conductance (CVC) were evaluated at four intradermal microdialysis sites with: 1) lactated Ringer (control), 2) nNOS inhibitor (nNOS-I, NPLA), 3) iNOS inhibitor (iNOS-I, 1400W), or 4) eNOS inhibitor (eNOS-I, LNAA). In younger adults during both exercise bouts, all inhibitors decreased sweating relative to control, albeit a lower sweat rate was observed at iNOS-I compared with eNOS-I and nNOS-I sites (all P < 0.05). CVC at the eNOS-I site was lower than control in younger adults throughout the intermittent exercise protocol (all P < 0.05). In older adults, there were no differences between control and iNOS-I sites for sweating and CVC during both exercise bouts (all P > 0.05). We show that iNOS and eNOS are the main contributors to NOS-dependent sweating and cutaneous vasodilation, respectively, in physically active younger adults exercising in the heat, and that iNOS inhibition does not alter sweating or cutaneous vasodilation in exercising physically active older adults.

Key pointsr Recent work demonstrates that nitric oxide (NO) contributes to cutaneous vasodilatation during moderate (400 W of metabolic heat production) but not high (700 W of metabolic heat production) intensity exercise bouts performed in the heat (35°C).r The present study evaluated whether the impairment in NO-dependent cutaneous vasodilatation was the result of a greater accumulation of reactive oxygen species during high (700 W of metabolic heat production) relative to moderate (500 W of metabolic heat production) intensity exercise.r It was shown that local infusion of ascorbate (an anti-oxidant) improves NO-dependent forearm cutaneous vasodilatation during high intensity exercise in the heat.r These findings provide novel insight into the physiological mechanisms governing cutaneous blood flow during exercise-induced heat stress and provide direction for future research exploring whether oxidative stress underlies the impairments in heat dissipation that may occur in older adults, as well as in individuals with pathophysiological conditions such as type 2 diabetes.Abstract Nitric oxide (NO)-dependent cutaneous vasodilatation is reportedly diminished during exercise performed at a high (700 W) relative to moderate (400 W) rate of metabolic heat production. The present study evaluated whether this impairment results from increased oxidative stress associated with an accumuluation of reactive oxygen species (ROS) during high intensity exercise. On two separate days, 11 young (mean ± SD, 24 ± 4 years) males cycled in the heat (35°C) at a moderate (500 W) or high (700 W) rate of metabolic heat production. Each session included two 30 min exercise bouts followed by 20 and 40 min of recovery, respectively. Cutaneous vascular conductance (CVC) was monitored at four forearm skin sites continuously perfused via intradermal microdialysis with: (1) lactated Ringer solution (Control); (2) 10 mM ascorbate (Ascorbate); (3) 10 mM L-NAME; or (4) 10 mM ascorbate + 10 mM L-NAME (Ascorbate + L-NAME). At the end of each 500 W exercise bout, CVC was attenuated with L-NAME (ß35% CVC max ) and Ascorbate + L-NAME (ß43% CVC max ) compared to Control (ß60% CVC max ; all P < 0.04); however, Ascorbate did not modulate CVC during exercise (ß60% CVC max ; both P > 0.87). Conversely, CVC was elevated with Ascorbate (ß72% CVC max ; both P < 0.03) but remained similar to Control (ß59% CVC max ) with L-NAME (ß50% CVC max ) and Ascorbate + L-NAME (ß47% CVC max ; all P > 0.05) at the end of both 700 W exercise bouts. We conclude that oxidative stress associated with an accumulation of ascorbate-sensitive ROS impairs NO-dependent cutaneous vasodilatation during intense exercise.

Population aging and global warming generate important public health risks, as older adults have increased susceptibility to heat stress (SHS). We defined and validated sex-specific screening criteria for SHS during work and leisure activities in hot environments in individuals aged 31-70 years using age, anthropometry, and cardiorespiratory fitness. A total of 123 males and 44 females [44 ± 14 years; 22.9 ± 7.4% body fat; 40.3 ± 8.6 peak oxygen uptake (mlO/kg/min)] participated, separated into the Analysis (n = 111) and Validation (n = 56) groups. Within these groups, participants were categorized into YOUNG (19-30 years; n = 47) and OLDER (31-70 years; n = 120). All participants performed exercise in the heat inside a direct calorimeter. Screening criteria for OLDER participants were defined from the Analysis group and were cross-validated in the Validation group. Results showed that 30% of OLDER individuals in the Analysis group were screened as SHS positive. A total of 274 statistically valid (p < 0.05) criteria were identified suggesting that OLDER participants were at risk for SHS when demonstrating two or more of the following (males/females): age ≥ 53.0/55.8 years; body mass index ≥29.5/25.7 kg/m; body fat percentage ≥ 28.8/34.9; body surface area ≤2.0/1.7 m; peak oxygen uptake ≤48.3/41.4 mlO/kg fat free mass/min. In the Validation group, McNemar χ comparisons confirmed acceptable validity for the developed criteria. We conclude that the developed criteria can effectively screen individuals 31-70 years who are at risk for SHS during work and leisure activities in hot environments and can provide simple and effective means to mitigate the public health risks caused by heat exposure.

Acetylcholine released from cholinergic nerves is involved in heat loss responses of cutaneous vasodilation and sweating. K(+) channels are thought to play a role in regulating cholinergic cutaneous vasodilation and sweating, though which K(+) channels are involved in their regulation remains unclear. We evaluated the hypotheses that 1) Ca(2+)-activated K(+) (KCa), ATP-sensitive K(+) (KATP), and voltage-gated K(+) (KV) channels all contribute to cholinergic cutaneous vasodilation; and 2) KV channels, but not KCa and KATP channels, contribute to cholinergic sweating. In 13 young adults (24 ± 5 years), cutaneous vascular conductance (CVC) and sweat rate were evaluated at intradermal microdialysis sites that were continuously perfused with: 1) lactated Ringer (Control), 2) 50 mM tetraethylammonium (KCa channel blocker), 3) 5 mM glybenclamide (KATP channel blocker), and 4) 10 mM 4-aminopyridine (KV channel blocker). At all sites, cholinergic cutaneous vasodilation and sweating were induced by coadministration of methacholine (0.0125, 0.25, 5, 100, and 2,000 mM, each for 25 min). The methacholine-induced increase in CVC was lower with the KCa channel blocker relative to Control at 0.0125 (1 ± 1 vs. 9 ± 6%max) and 5 (2 ± 5 vs. 17 ± 14%max) mM methacholine, whereas it was lower in the presence of KATP (69 ± 7%max) and KV (57 ± 14%max) channel blocker compared with Control (79 ± 6%max) at 100 mM methacholine. Furthermore, methacholine-induced sweating was lower at the KV channel blocker site (0.42 ± 0.17 mg·min(-1)·cm(-2)) compared with Control (0.58 ± 0.15 mg·min(-1)·cm(-2)) at 2,000 mM methacholine. In conclusion, we show that KCa, KATP, and KV channels play a role in cholinergic cutaneous vasodilation, whereas only KV channels contribute to cholinergic sweating in normothermic resting humans.

Many studies have aimed to identify the controllers of sweating using ventilated capsules with intradermal microdialysis. It is unclear, however, if the surface area covered by the capsule influences the observed response as a result of differences in the number of sweat glands affected by the infused pharmacological agent relative to the total glands captured by the capsule. We evaluated the area of skin perfused with agents delivered via microdialysis. Thereafter, we developed a specialized sweat capsule (1.1 cm2) and compared the sweating response with a classic capsule (2.8 cm2). In Protocol 1 (n = 6), methacholine was delivered to forearm skin in a dose‐dependent manner (1–2000 mmol L−1). The area of activated sweat glands was assessed via the modified iodine‐paper technique. In Protocol 2 (n = 6), the area of inhibited sweat glands induced by ouabain and atropine was assessed during moderate‐intensity cycling. Marked variability in the affected skin area was observed (0.9 ± 0.4 to 5.2 ± 1.1 cm2). In Protocol 3 (n = 6), we compared the attenuation in local sweat rate (LSR) induced by atropine between the new and classic capsule during moderate‐intensity cycling. Atropine attenuated sweating as assessed using the new (control: 0.87 ± 0.23 mg min−1 cm−2 vs. atropine: 0.54 ± 0.22 mg min−1 cm−2; P < 0.01) and classic (control: 0.85 ± 0.33 mg min−1 cm−2 vs. atropine: 0.60 ± 0.26 mg min−1 cm−2; P = 0.05) capsule designs. Importantly, responses did not differ between capsule designs (P = 0.23). These findings provide critical information regarding the skin surface area perfused by microdialysis and suggest that use of a larger capsule does not alter the mechanistic insight into the sweating response gained when using microdialysis.

https://mc06.manuscriptcentral.com/apnm-pubs

Key pointsr Nitric oxide synthase (NOS) contributes to sweating and cutaneous vasodilatation during exercise in the heat.r Similarly, reports show that Na + /K + -ATPase activation can modulate sweating and microvascular circulation. In light of the fact that NO can activate Na + /K + -ATPase, we evaluated whether there is an interaction between Na + /K + -ATPase and NOS in the regulation of heat loss responses during an exercise-induced heat stress.r We demonstrate that Na + /K + -ATPase and NOS do not synergistically influence local forearm sweating during moderate intensity (fixed rate of metabolic heat production of 500 W) exercise in the heat (35°C). Conversely, we show an interactive role between NOS and Na + /K + -ATPase in the modulation of cutaneous vasodilatation.r These findings provide novel insight regarding the mechanisms underpinning the control of sweating and cutaneous vasodilatation during exercise in the heat. Given that ouabain may be prescribed as a cardiac glycoside in clinical settings, potential heat loss impairments with ouabain administration should be explored.Abstract Nitric oxide (NO) synthase (NOS) contributes to the heat loss responses of sweating and cutaneous vasodilatation. Given that NO can activate Na + /K + -ATPase, which also contributes to sweating and microvasculature regulation, we evaluated the separate and combined influence of Na + /K + -ATPase and NOS on sweating and cutaneous vasodilatation. Thirteen young (23±3 years) males performed two 30 min semi-recumbent cycling bouts in the heat (35°C) at a fixed rate of metabolic heat production (500 W) followed by 20 and 40 min recoveries, respectively. Local sweat rate (LSR) and cutaneous vascular conductance (CVC) were measured at four forearm skin sites continuously perfused via intradermal microdialysis with either: (1) lactated Ringer solution (Control); (2) 6 mᴍ ouabain (Ouabain), a Na + /K + -ATPase inhibitor; (3) 10 mᴍ L-N G -nitroarginine methyl ester (L-NAME), a NOS inhibitor; or (4) 6 mᴍ ouabain and 10 mᴍ L-NAME (Ouabain+L-NAME). At the end of both exercise bouts relative to Control, LSR was attenuated with Ouabain (54-60%), L-NAME (12-13%) and Ouabain+L-NAME (68-74%; all P < 0.05). Moreover, the sum of attenuations from Control induced by independent administration of Ouabain and L-NAME was similar to the combined infusion of Ouabain+L-NAME (both P ࣙ 0.74). Compared to Control, CVC at the end of both exercise bouts was similar with Ouabain (both P ࣙ 0.30), but attenuated with L-NAME (%CVC max reduction from Control, 24-25%). Furthermore, CVC at the Ouabain+L-NAME site (38-39%; all P < 0.01) was attenuated compared to Control and did not differ from baseline resting values (both P ࣙ 0.81). We show that Na + /K + -ATPase and NOS do not synergistically mediate sweating, whereas they influence cutaneous blood flow in an interactive manner during exercise in the heat.

What is the central question of this study? Aerobic fitness modulates heat loss, but the heat-load threshold at which fitness-related differences in heat loss occur in young healthy men remains unclear. What is the main finding and its importance? We demonstrate using direct calorimetry that aerobic fitness modulates heat loss in a heat-load-dependent manner, with fitness-related differences occurring between young men who have low and high fitness when the heat load is ∼≥500 W. Although aerobic fitness has been known for some time to modulate heat loss, our findings define the precise heat-load threshold at which fitness-related differences occur. The effect of aerobic fitness (defined as rate of peak oxygen consumption) on heat loss during exercise is thought to be related to the level of heat stress. However, it remains unclear at what combined exercise and environmental (net) heat-load threshold these fitness-related differences occur. To identify this, we assessed whole-body heat exchange (dry and evaporative) by direct calorimetry in young (22 ± 3 years) men matched for physical characteristics with low (Low-fit; 39.8 ± 2.5 ml O kg min ), moderate (Mod-fit; 50.9 ± 1.2 ml O kg min ) and high aerobic fitness (High-fit; 62.0 ± 4.4 ml O kg min ; each n = 8), during three 30 min bouts of cycling in dry heat (40°C, 12% relative humidity) at increasing rates of metabolic heat production of 300 (Ex1), 400 (Ex2) and 500 W (Ex3), each followed by a 15 min recovery period. Each group was exposed to a similar net heat load (metabolic plus ∼100 W dry heat gain; P = 0.83) during each exercise bout [∼400 (Ex1), ∼500 (Ex2) and ∼600 W (Ex3); P < 0.01]. Although evaporative heat loss was similar between groups during Ex1 (P = 0.33), evaporative heat loss was greater in the High-fit (Ex2, 466 ± 21 W; Ex3, 557 ± 26 W) compared with the Low-fit group (Ex2, 439 ± 22 W; Ex3, 511 ± 20 W) during Ex2 and Ex3 (P ≤ 0.03). Conversely, evaporative heat loss for the Mod-fit group did not differ from either the High-fit or Low-fit group during all exercise bouts (P ≥ 0.09). We demonstrate that aerobic fitness modulates heat loss in a heat-load-dependent manner, such that young, highly fit men display greater heat-loss capacity only at heat loads ∼≥500 W compared with their lesser trained counterparts.