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Aims/hypothesis: Early compromised endothelial function challenges the ability of individuals with type 1 diabetes to perform normal physical exercise. The exact mechanisms underlying this vascular limitation remain unknown, but may involve either formation or metabolism of nitric oxide (NO), a major vasodilator, whose activity is known to be compromised by oxidative stress., Methods: Muscle microvascular reactivity (near-infrared spectroscopy) to an incremental exhaustive bout of exercise was assessed in 22 adults with uncomplicated type 1 diabetes (HbA 1c 64.5 ± 15.7 mmol/mol; 8.0 ± 1.4%) and in 21 healthy individuals (18-40 years of age). NO-related substrates/metabolites were also measured in the blood along with other vasoactive compounds and oxidative stress markers; measurements were taken at rest, at peak exercise and after 15 min of recovery. Demographic characteristics, body composition, smoking status and diet were comparable in both groups., Results: Maximal oxygen uptake was impaired in individuals with type 1 diabetes compared with in healthy participants (35.6 ± 7.7 vs 39.6 ± 6.8 ml min -1  kg -1 , p < 0.01) despite comparable levels of habitual physical activity (moderate to vigorous physical activity by accelerometery, 234.9 ± 160.0 vs 280.1 ± 114.9 min/week). Compared with non-diabetic participants, individuals with type 1 diabetes also displayed a blunted exercise-induced vasoreactivity (muscle blood volume at peak exercise as reflected by ∆ total haemoglobin, 2.03 ± 5.82 vs 5.33 ± 5.54 μmol/l; interaction 'exercise' × 'group', p < 0.05); this was accompanied by lower K + concentration (p < 0.05), reduced plasma L-arginine (p < 0.05)-in particular when HbA 1c was high (mean estimation: -4.0, p < 0.05)-and lower plasma urate levels (p < 0.01). Nonetheless, exhaustive exercise did not worsen lipid peroxidation or other oxidative stress biomarkers, and erythrocytic enzymatic antioxidant resources were mobilised to a comparable extent in both groups. Nitrite and total nitrosation products, which are potential alternative NO sources, were similarly unaltered. Graphical abstract CONCLUSIONS/INTERPRETATION: Participants with uncomplicated type 1 diabetes displayed reduced availability of L-arginine, the essential substrate for enzymatic nitric oxide synthesis, as well as lower levels of the major plasma antioxidant, urate. Lower urate levels may reflect a defect in the activity of xanthine oxidase, an enzyme capable of producing NO from nitrite under hypoxic conditions. Thus, both canonical and non-canonical NO production may be reduced. However, neither of these changes exacerbated exercise-induced oxidative stress., Trial Registration: clinicaltrials.gov NCT02051504.

Nitric oxide (NO) is a gaseous signaling molecule that plays an important role in myriad physiological processes, including the regulation of vascular tone, neurotransmission, mitochondrial respiration, and skeletal muscle contractile function. NO may be produced via the canonical NO synthase-catalyzed oxidation of l-arginine and also by the sequential reduction of nitrate to nitrite and then NO. The body's nitrate stores can be augmented by the ingestion of nitrate-rich foods (primarily green leafy vegetables). NO bioavailability is greatly enhanced by the activity of bacteria residing in the mouth, which reduce nitrate to nitrite, thereby increasing the concentration of circulating nitrite, which can be reduced further to NO in regions of low oxygen availability. Recent investigations have focused on promoting this nitrate-nitrite-NO pathway to positively affect indices of cardiovascular health and exercise tolerance. It has been reported that dietary nitrate supplementation with beetroot juice lowers blood pressure in hypertensive patients, and sodium nitrite supplementation improves vascular endothelial function and reduces the stiffening of large elastic arteries in older humans. Nitrate supplementation has also been shown to enhance skeletal muscle function and to improve exercise performance in some circumstances. Recently, it has been established that nitrate concentration in skeletal muscle is much higher than that in blood and that muscle nitrate stores are exquisitely sensitive to dietary nitrate supplementation and deprivation. In this review, we consider the possibility that nitrate represents an essential storage form of NO and discuss the integrated function of the oral microbiome, circulation, and skeletal muscle in nitrate-nitrite-NO metabolism, as well as the practical relevance for health and performance., (Copyright © 2020 by the American College of Sports Medicine.)

Introduction: Nitric oxide (NO) precursor supplementation has been shown to increase NO bioavailability and can potentially improve vascular function and exercise performance. It remains unclear whether the combination of NO precursor supplementation and exercise training has synergic effects on exercise performance. This study aims to assess the effect of chronic nitrate and citrulline intake on exercise training adaptations in healthy young individuals., Methods: In this randomized, double-bind trial, 24 healthy young (12 females) subjects performed vascular function assessment (blood pressure, pulse wave velocity, postischemia vasodilation, and cerebrovascular reactivity) and both local (submaximal isometric unilateral knee extension) and whole-body (incremental cycling) exercise tests to exhaustion before and after a 2-month exercise training program and daily intake of a placebo or a nitrate-rich salad and citrulline (N + C, 520 mg nitrate and 6 g citrulline) drink. Prefrontal cortex and quadriceps oxygenation was monitored continuously during exercise by near-infrared spectroscopy., Results: N + C supplementation had no effect on vascular function and muscle and cerebral oxygenation during both local and whole-body exercise. N + C supplementation induced a significantly larger increase in maximal knee extensor strength (+5.1 ± 3.5 vs +0.2 ± 5.5 kg, P = 0.008) as well as a trend toward a larger increase in knee extensor endurance (+35.2 ± 26.1 vs +24.0 ± 10.4 contractions, P = 0.092) than placebo, but no effect on exercise training-induced maximal aerobic performance improvement., Conclusion: These results suggest that chronic nitrate and citrulline supplementation enhances the effect of exercise training on quadriceps muscle function in healthy active young individuals, but this does not translate into improved maximal aerobic performances.

This study aimed to clarify whether muscle-derived irisin secretion induced by aerobic exercise training is involved in reduction of arterial stiffness via arterial nitric oxide (NO) productivity in obesity. In animal study, 16 Otsuka Long-Evans Tokushima Fatty (OLETF) rats with obesity were randomly divided into 2 groups: sedentary control (OLETF-CON) and 8-week aerobic treadmill training (OLETF-EX) groups. In human study, 15 subjects with obesity completed 8-week aerobic exercise training for 45 min at 60%-70% peak oxygen uptake intensity for 3 days/week. As a result of animal study, carotid-femoral pulse wave velocity (cfPWV) was decreased, and arterial phosphorylation levels of AMP-activated protein kinase (AMPK), protein kinase B (Akt), and endothelial NO synthase (eNOS), circulating levels of nitrite/nitrate (NOx) and irisin, and muscle messenger RNA expression of fibronectin type III domain containing 5 ( Fndc5 ) were increased in the OLETF-EX group compared with OLETF-CON group. In a human study, regular aerobic exercise reduced cfPWV and elevated circulating levels of NOx and irisin. Furthermore, change in circulating irisin levels by regular exercise was positively correlated with circulating NOx levels and was negatively correlated with cfPWV. Thus, aerobic exercise training-induced increase in irisin secretion may be related to reduction of arterial stiffness achieved by NO production via activated arterial AMPK-Akt-eNOS signaling pathway in obesity. Novelty Aerobic exercise training promoted irisin secretion with upregulation of muscle Fndc5 gene expression in rats with obesity. Irisin affected the activation of arterial AMPK-Akt-eNOS signaling by aerobic exercise training. Increased serum irisin level by aerobic exercise training was associated with reduction of arterial stiffness in obese adults.

Aim: To investigate the influence of the intranasal AlaxoLito Plus Nasal Stent during exercise on nitric oxide (NO) synthesis, NO exhalation, red blood cell (RBC) deformability and oxygen uptake., Methods: Parameters were measured before and after acute cycle ergometer test at different intensities. Spirometric, microrheological and NO parameters were determined for oral (OB), nasal (NB) and nasal-stent breathing (SB). RBC deformability was measured and elongation indices for 3.87 Pa and maximal deformability were calculated. RBC/plasma/exhaled NO, oxygen uptake and respiratory rate were determined., Results: Exhaled NO was higher at rest during OB compared to SB and NB and decreased after exercise with NB and SB. Plasma and RBC NO remained unaltered during intervention. RBC deformability increased at moderate intensity during SB. Deformability decreased at moderate and medium intensity with NB. Respiratory rate for same oxygen uptake did not differ between breathing settings., Conclusion: The AlaxoLito Plus Nasal Stent may modulate deformability during moderate exercise and increase NO exhalation without major effects on oxygen uptake and performance., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Glutathione (GSH) is the main non-enzymatic antioxidant playing an important role in detoxification, signal transduction by modulation of protein thiols redox status and direct scavenging of radicals. The latter function is not only performed against reactive oxygen species (ROS) but GSH also has a fundamental role in buffering nitric oxide (NO), a physiologically-produced molecule having-multifaceted functions. The efficient rate of GSH synthesis and high levels of GSH-dependent enzymes are characteristic features of healthy skeletal muscle where, besides the canonical functions, it is also involved in muscle contraction regulation. Moreover, NO production in skeletal muscle is a direct consequence of contractile activity and influences several metabolic myocyte pathways under both physiological and pathological conditions. In this review, we will consider the homeostasis and intersection of GSH with NO and then we will restrict the discussion on their role in processes related to skeletal muscle function and degeneration.

Purpose: Physical activity is an important part of life, and exercise-induced asthma (EIA) can reduce the quality of life. A standardized exercise challenge is needed to diagnose EIA, but this is a time consuming, effortful and expensive method. Exhaled nitric oxide (eNO) as a marker of eosinophil inflammation is determined rapidly and easily. The aim of this study was to investigate eNO as surrogate marker for predicting a positive reaction in an exercise challenge in a cold chamber (ECC)., Methods: A total of 143 subjects aged 6-45 years with suspected EIA were recruited for the study. The subjects underwent an eNO measurement, an ECC and a skin prick test (SPT). To define the sensitivity and specificity of eNO as predictor, a receiver-operating characteristic (ROC) curve was plotted. The individual probability of the occurrence of a positive reaction after ECC based on an eNO value was calculated using a logistic regression model., Results: An eNO cut-off value of 18.5 ppb (area under the curve (AUC) 0.71, p < 0.001) showed the best combination of sensitivity and specificity for a positive reaction (forced expiratory volume in 1 s (FEV 1 ) decrease ≥ 10% after ECC) for the whole group. An eNO cut-off value of 46.0 ppb had a specificity of 100.0% to predict a significant FEV 1 decrease and may save exercise testing in 22.4% of patients. A negative predictive level with a high sensitivity and negative predictive value (NPV) could not be defined. In the subgroup that was house dust might (HDM) allergy positive (HDM pos; n = 68, 45.5% of all subjects), an eNO cut-off value of 35.5 ppb (AUC 0.79, p < 0.01) showed the best combination of sensitivity and specificity for a positive reaction after the ECC with a specificity 100.0% and may save exercise testing in 45.6% of HDM pos patients. Using logistic regression, a 95% probability for a positive FEV 1 decrease after ECC was estimated at 53 ppb for the whole group and at 47 ppb for the HDM pos subgroup., Conclusions: Exhaled NO measurement is a screening tool for EIA, especially in HDM pos subjects. In a real-life setting, a cut-off value of 46.0 ppb detects EIA at 100% in all suspected patients, and a cut-off level of 35.5 ppb is valuable marker of EIA in patients with an HDM allergy. These levels can save time and costs in a large proportion of patients and will be helpful for clinicians., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Purpose: Previous work has shown nitric oxide (NO) contributes to ~15% of the hyperemic response to dynamic exercise in healthy humans. This NO-mediated vasodilation occurs, in part, via increases in intracellular cyclic guanosine monophosphate (cGMP), which is catabolized by phosphodiesterase. We sought to examine the effect of phosphodiesterase-5 (PDE-5) inhibition on forearm blood flow (FBF) responses to dynamic handgrip exercise in healthy humans and the role of NO. We hypothesized exercise hyperemia would be augmented by sildenafil citrate (SDF, PDE-5 inhibitor). We further hypothesized any effect of SDF on exercise hyperemia would be abolished with intra-arterial infusion of the NO synthase (NOS) inhibitor L-N G -monomethyl arginine (L-NMMA)., Methods: FBF (Doppler ultrasound) was assessed at rest and during 5 min of dynamic forearm handgrip exercise at 15% of maximal voluntary contraction under control (saline) conditions and during 3 experimental protocols: (1) oral SDF (n = 10), (2) intra-arterial L-NMMA (n = 20), (3) SDF and L-NMMA (n = 10). FBF responses to intra-arterial sodium nitroprusside (NTP, NO donor) were also assessed., Results: FBF increased with exercise (p < 0.01). Intra-arterial infusion of L-NMMA resulted in a reduction in exercise hyperemia (17 ± 1 to 15 ± 1 mL/dL/min, p < 0.01). Although the hyperemic response to NTP was augmented by SDF (area under the curve: 41 ± 7 vs 61 ± 11 AU, p < 0.01), there was no effect of SDF on exercise hyperemia (p = 0.33)., Conclusions: Despite improving NTP-mediated vasodilation, oral SDF failed to augment exercise hyperemia in young, healthy adults. These observations reflect a minor contribution of NO and the cGMP pathway during exercise hyperemia in healthy young humans.

Background: The mechanisms underlying red blood cell (RBC)-mediated hypoxic vasodilation remain controversial, with separate roles for nitrite () and S-nitrosohemoglobin (SNO-Hb) widely contested given their ability to transduce nitric oxide bioactivity within the microcirculation. To establish their relative contribution in vivo, we quantified arterial-venous concentration gradients across the human cerebral and femoral circulation at rest and during exercise, an ideal model system characterized by physiological extremes of O 2 tension and blood flow., Methods: Ten healthy participants (5 men, 5 women) aged 24±4 (mean±SD) years old were randomly assigned to a normoxic (21% O 2 ) and hypoxic (10% O 2 ) trial with measurements performed at rest and after 30 minutes of cycling at 70% of maximal power output in hypoxia and equivalent relative and absolute intensities in normoxia. Blood was sampled simultaneously from the brachial artery and internal jugular and femoral veins with plasma and RBC nitric oxide metabolites measured by tri-iodide reductive chemiluminescence. Blood flow was determined by transcranial Doppler ultrasound (cerebral blood flow) and constant infusion thermodilution (femoral blood flow) with net exchange calculated via the Fick principle., Results: Hypoxia was associated with a mild increase in both cerebral blood flow and femoral blood flow (P<0.05 versus normoxia) with further, more pronounced increases observed in femoral blood flow during exercise (P<0.05 versus rest) in proportion to the reduction in RBC oxygenation (r=0.680-0.769, P<0.001). Plasma gradients reflecting consumption (arterial>venous; P<0.05) were accompanied by RBC iron nitrosylhemoglobin formation (venous>arterial; P<0.05) at rest in normoxia, during hypoxia (P<0.05 versus normoxia), and especially during exercise (P<0.05 versus rest), with the most pronounced gradients observed across the bioenergetically more active, hypoxemic, and acidotic femoral circulation (P<0.05 versus cerebral). In contrast, we failed to observe any gradients consistent with RBC SNO-Hb consumption and corresponding delivery of plasma S-nitrosothiols (P>0.05)., Conclusions: These findings suggest that hypoxia and, to a far greater extent, exercise independently promote arterial-venous delivery gradients of intravascular nitric oxide, with deoxyhemoglobin-mediated reduction identified as the dominant mechanism underlying hypoxic vasodilation., (© 2016 American Heart Association, Inc.)

Background: Acute exercise increases red blood cell-nitric oxide synthase (RBC-NOS) activation and RBC deformability but the effect of regular training remains unclear., Objective: To detect the chronic effect of enduring moderate and high intensity training on the RBC-NOS/NO pathway and to detect a relation between RBC deformability and endurance capacity., Methods: 38 healthy male subjects were randomly assigned to one of three training groups: High Volume Training (HVT; 120-140 beats per minute (bpm)), High Intensity Training (HIT; 160-180 bpm) and Moderate Intensity Training (MIT; 140-160 bpm). Blood parameters, maximum oxygen capacity (VO2 max), RBC deformability, RBC nitrite level and RBC-NOS activation were measured after venous blood sampling at rest pre (T0) and after six weeks of training (T1)., Results: RBC-NOS activation, RBC nitrite concentration and RBC deformability were significantly increased at T1 in the HIT group. Parameters were unaltered in MIT and HVT. Maximum oxygen uptake was only significantly increased in the HIT group and regression analysis revealed positive regression between VO2 max and RBC deformability., Conclusions: High intensity training was the only training programme that sustainably affected RBC-NOS dependent NO production and performance capacity. HIT therefore represents a time efficient training program resulting in improved RBC function potentially improving physical condition.

Nitric oxide (NO) is a small molecule implicated in multiple signal transduction pathways thus contributing to the regulation of many cellular functions. The identification of NO synthase (NOS) isoforms and the subsequent characterization of the mechanisms of cell activation of the enzymes permitted the partial understanding of both the physiological and pathological processes. NO bioavailability plays an important role in the pathophysiology of cardiovascular disease and its reduction in endothelial cells is strictly associated to endothelial dysfunction which, in turn, correlates with cardiovascular mortality. Indeed, endothelial NO synthase (eNOS) has a key role in limiting cardiac dysfunction and remodeling in heart diseases, in part by decreasing myocyte hypertrophy. Conversely, exercise training is recommended to prevent and treat cardiovascular diseases-associated disorders at least by enhanced NO synthase activity and expression, and increased production of antioxidants, which prevents premature breakdown of NO. Exercise training may cause an improvement in endothelial function for both experimental animals and humans; Studies in both healthy subjects and patients with impaired NO-related vasorelaxation remarked exercise training ability to improve vascular structure and function and endothelial homeostasis. This chapter will briefly consider the importance of NO signaling in the maintenance of cardiovascular physiology, and discuss recent insights into the effect of exercise training on the signaling pathways that modulate NO synthesis and degradation in health and cardiovascular disease. In addition, we will highlight the molecular mechanisms via which microRNAs (miRs) target NO signaling in the cardiovascular system, and NO as a candidate molecule for development of new therapies.

Growing evidence has shown that acute exercise impairs erythrocyte membrane structure and function as a consequence of increased physical and chemical stress. Erythrocyte-synthesized nitric oxide (NO) is known to modulate membrane fluidity, and its bioavailability depends on the balance between its production and scavenging by reactive oxygen species. Here, we investigated whether a maximal exercise test could affect erythrocyte NO bioavailability and oxidative stress. Twelve men (26±4 years old, V̇O2peak 44.1±4.3 mL·kg-1·min-1) performed a treadmill maximal cardiopulmonary exercise test. Blood was collected at rest and immediately after exercise for erythrocytes isolation. Maximal exercise caused an increase in erythrocytes count, haemoglobin and haematocrit levels. There was no change in L-arginine influx into erythrocytes after exercise. Yet, nitric oxide synthase activity, and thus, NO production, was increased after maximal test, as well cyclic GMP levels. In relation to biomarkers of oxidative stress, maximal test resulted in increased levels of lipid peroxidation, and diminished superoxide dismutase activity. Neither glutathione peroxidase nor catalase activity was affected by maximal test. Our findings demonstrate that the increased erythrocyte membrane rigidity caused by an acute bout of exercise may be caused, in part, by an increased lipid oxidative damage caused by ROS produced exogenously.

Both cyclooxygenase (COX) and nitric oxide synthase (NOS) contribute to sweating, whereas NOS alone contributes to cutaneous vasodilation during exercise in the heat. Here, we evaluated if Type 1 diabetes mellitus (T1DM) modulates these responses. Adults with (n = 11, 25 ± 5 yr) and without (n = 12, 24 ± 4 yr) T1DM performed two bouts of 30-min cycling at a fixed rate of heat production of 400 W in the heat (35°C); each followed by a 20- and 40-min recovery period, respectively. Sweat rate and cutaneous vascular conductance (CVC) were measured at four intradermal microdialysis sites treated with either 1) lactated Ringer (vehicle control site), 2) 10 mM ketorolac (nonselective COX inhibitor), 3) 10 mM N G -nitro-l-arginine methyl ester (nonselective NOS inhibitor), or 4) a combination of both inhibitors. In nondiabetic adults, separate and combined inhibition of COX and NOS reduced exercise sweat rate (P ≤ 0.05), and the magnitude of reductions were similar across sites. In individuals with T1DM, inhibition of COX resulted in an increase in sweat rate of 0.10 ± 0.09 and 0.09 ± 0.08 mg ·: min -1 ·: cm -2 for the first and second exercise bouts, respectively, relative to vehicle control site (P ≤ 0.05), whereas NOS inhibition had no effect on sweating. In both groups, NOS inhibition reduced CVC during exercise (P ≤ 0.05), although the magnitude of reduction did not differ between the nondiabetic and T1DM groups (exercise 1: -28 ± 10 vs. -23 ± 8% max, P = 0.51; exercise 2: -31 ± 12 vs. -24 ± 10% max, P = 0.38). We show that in individuals with T1DM performing moderate intensity exercise in the heat, NOS-dependent sweating but not cutaneous vasodilation is attenuated, whereas COX inhibition increases sweating., (Copyright © 2016 the American Physiological Society.)

Although dietary nitrate (NO 3 ) ingestion appears to enhance exercise capacity and performance in young individuals, inconclusive findings have been reported in older people. Therefore, we conducted a double-blind, crossover randomized clinical trial using beetroot juice in older healthy participants, who were classified as normal weight and overweight. We tested whether consumption of beetroot juice (a rich source of NO - ) for 1 week would increase nitric oxide bioavailability via the nonenzymatic pathway and enhance (1) exercise capacity during an incremental exercise test, (2) physical capability, and (3) free-living physical activity. Twenty nonsmoking, healthy participants between 60 and 75 years of age and with a body mass index of 20.0 to 29.9 kg/m 3 - ) for 1 week would increase nitric oxide bioavailability via the nonenzymatic pathway and enhance (1) exercise capacity during an incremental exercise test, (2) physical capability, and (3) free-living physical activity. Twenty nonsmoking, healthy participants between 60 and 75 years of age and with a body mass index of 20.0 to 29.9 kg/m 2 concentrations (P<.001) when compared with placebo. Beetroot juice did not influence resting or submaximal and maximal oxygen consumption during the incremental exercise test. In addition, measures of physical capability and physical activity levels measured in free-living conditions were not modified by beetroot juice ingestion. The positive effects of beetroot juice ingestion on exercise performance seen in young individuals were not replicated in healthy, older adults. Whether aging represents a modifier of the effects of dietary NO 3 - concentrations were measured by gas chromatography-mass spectrometry. Nineteen participants (male-to-female ratio, 9:10) completed the study.Beetroot juice increased significantly both plasma and urinary NO 3 - concentrations (P<.001) when compared with placebo. Beetroot juice did not influence resting or submaximal and maximal oxygen consumption during the incremental exercise test. In addition, measures of physical capability and physical activity levels measured in free-living conditions were not modified by beetroot juice ingestion. The positive effects of beetroot juice ingestion on exercise performance seen in young individuals were not replicated in healthy, older adults. Whether aging represents a modifier of the effects of dietary NO 3 - on muscular performance is not known, and mechanistic studies and larger trials are needed to test this hypothesis., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Both endothelial nitric oxide synthase (eNOS) gene polymorphism and nitric oxide (NO) are involved in important cardiovascular, muscular and inflammatory physiological mechanisms during ageing and response to exercise. The aim of this study was to investigate the NO kinetic response following an acute eccentric resistance exercise (ERE) session and the possible effect of the Glu298Asp eNOS gene polymorphism in elderly obese women. Eighty-seven women (age 69·4 ± 6·1 years, body weight 74·9 ± 12·7 kg, height 151·9 ± 6·0 cm and BMI 32·5 ± 5·7 kg m -2 ) completed seven sets of ten eccentric repetitions at 110% of the ten repetitions maximum (10RM). NO concentrations remained elevated up to 48 h following the acute ERE session as compared with baseline, for GG and GT/TT groups (P<0·05), with no differences between genotypes. The GG genotype group had higher body weight, prevalence of obesity (BMI classification - 81% versus 56%), BMI and higher relative muscle strength, while they had significantly lower triglycerides, VLDL and urea concentrations as compared with TT/TG group. In conclusion, NO remains elevated for up to 48 h after an acute ERE session, without genotype interaction. The TT/TG genotype had a negative impact on triglycerides, VLDL and urea concentrations. Thus, T carriers should increase their attention to cardiovascular risk factor and metabolic disorders., (© 2015 Scandinavian Society of Clinical Physiology and Nuclear Medicine. Published by John Wiley & Sons Ltd.)

It is becoming increasingly apparent that a high vasodilator response of the skeletal muscle microvasculature to insulin and exercise is of critical importance for adequate muscle perfusion and long-term microvascular and muscle metabolic health. Previous research has shown that a sedentary lifestyle, obesity and ageing lead to impairments in the vasodilator response, while a physically active lifestyle keeps both microvascular density and vasodilator response high. To investigate the molecular mechanisms behind these impairments and the benefits of exercise training interventions, our laboratory has recently developed quantitative immunofluorescence microscopy methods to measure protein content of eNOS and NAD(P)Hoxidase specifically in the endothelial layer of capillaries and arterioles of human skeletal muscle. As eNOS produces nitric oxide (NO) and NAD(P)Hoxidase produces superoxide anions (O2 (-) , quenching NO) we propose that the eNOS/NAD(P)Hoxidase protein ratio is a marker of vasodilator capacity. The novel methods show that endurance training (ET) and high intensity interval training (HIT), generally regarded as a time-efficient alternative to ET, increase eNOS protein content and the eNOS/NADP(H)oxidase protein ratio in previously sedentary lean and obese young men. Resistance exercise training had smaller but qualitatively similar effects. Western blot data of other laboratories suggest that endurance exercise training leads to similar changes in sedentary elderly men. Future research will be required to investigate the relative importance of other sources and tissues in the balance between NO and O2 (-) production seen by the vascular smooth muscle layer of terminal arterioles., (© 2015 The Authors. The Journal of Physiology © 2015 The Physiological Society.)

Asthmatic patients with fixed airway obstruction (FAO) and patients with chronic obstructive pulmonary disease (COPD) share similarities in terms of irreversible pulmonary function impairment. Exhaled nitric oxide (eNO) has been documented as a marker of airway inflammation in asthma, but not in COPD. To examine whether the basal eNO level and the change after exercise may differentiate asthmatics with FAO from COPD, 27 normal subjects, 60 stable asthmatics, and 62 stable COPD patients were studied. Asthmatics with FAO (n = 29) were defined as showing a postbronchodilator FEV1/forced vital capacity (FVC) ≤70% and FEV1 less than 80% predicted after inhaled salbutamol (400 μg). COPD with dynamic hyperinflation (n = 31) was defined as a decrease in inspiratory capacity (ΔIC%) after a 6 minute walk test (6MWT). Basal levels of eNO were significantly higher in asthmatics and COPD patients compared to normal subjects. The changes in eNO after 6MWT were negatively correlated with the percent change in IC (r = -0.380, n = 29, P = 0.042) in asthmatics with FAO. Their levels of basal eNO correlated with the maximum mid-expiratory flow (MMEF % predicted) before and after 6MWT. In COPD patients with air-trapping, the percent change of eNO was positively correlated to ΔIC% (rs = 0.404, n = 31, P = 0.024). We conclude that asthma with FAO may represent residual inflammation in the airways, while dynamic hyperinflation in COPD may retain NO in the distal airspace. eNO changes after 6MWT may differentiate the subgroups of asthma or COPD patients and will help toward delivery of individualized therapy for airflow obstruction.

Background: Fractional expired nitric oxide (FENO ) is decreased after exercise. The effect of exercise in the cold upon FENO is unknown., Purpose: To examine changes in FENO after a short, high intensive exercise test in a cold and in a temperate environment., Methods: Twenty healthy well-trained subjects (eight females) aged 18-28 years performed an 8-min exercise test at 18°C (SD = 1.0) and -10°C (SD = 1.2) ambient temperature. The tests were performed in a climate chamber in random order. The workload corresponded to 90-95% of peak heart rate (HRpeak ) during the last 4 min. FENO was measured offline. Exhaled gas was sampled in Mylar(®) bags using a collector kit with a flow restrictor and analysed within 2 h. FENO was measured before exercise and repeatedly during the first hour after. ANOVA for repeated measures was used to compare differences in FENO after exercise between environments., Results: There was no difference in baseline FENO . A significant difference in FENO between environments was found after warm-up and from 20 to 30 min after exercise, with FENO being lower after exercise in the cold (P<0.05). The maximal reduction in FENO was seen 5 min after exercise and was not different between environments., Conclusion: Recovery of FENO was slower after exercising in -10°C compared with 18°C., (© 2014 Scandinavian Society of Clinical Physiology and Nuclear Medicine. Published by John Wiley & Sons Ltd.)

Ageing is associated with an increased risk of developing noncommunicable diseases (NCDs), such as diabetes and cardiovascular disease (CVD). The increased risk can be attributable to increased prolonged exposure to oxidative stress. Often, CVD is preceded by endothelial dysfunction, which carries with it a proatherothrombotic phenotype. Endothelial senescence and reduced production and release of nitric oxide (NO) are associated with "vascular ageing" and are often accompanied by a reduced ability for the body to repair vascular damage, termed "reendothelialization." Exercise has been repeatedly shown to confer protection against CVD and diabetes risk and incidence. Regular exercise promotes endothelial function and can prevent endothelial senescence, often through a reduction in oxidative stress. Recently, endothelial precursors, endothelial progenitor cells (EPC), have been shown to repair damaged endothelium, and reduced circulating number and/or function of these cells is associated with ageing. Exercise can modulate both number and function of these cells to promote endothelial homeostasis. In this review we look at the effects of advancing age on the endothelium and these endothelial precursors and how exercise appears to offset this "vascular ageing" process.

Objective: Fractional exhaled nitric oxide (FENO) and exercise testing are widely used for the evaluation of pediatric asthma. The evidence relating to the effects of strenuous exercise on FENO in children is conflicting. Little information is available on the association between exercise and FENO in relation to allergic rhinoconjunctivitis (AR). We aimed to investigate the effects of AR on children's FENO in response to a standardized treadmill exercise test., Methods: A total of 124 children with current asthma and 124 non-asthmatic children aged 8-16 years were studied. FENO was measured at baseline, at 1 and 30 min after an exercise challenge test using the single breath technique with EcoMedics Exhalyzer. A structured parental interview, spirometry, serum allergen-specific IgE and skin prick tests were performed., Results: Baseline FENO was higher in both asthmatics and non-asthmatics with AR than without AR (both p < 0.001). The FENO time trend was dependent on AR (p = 0.039), irrespective of asthma (p = 0.876). In children with AR, FENO had declined at 1 min by a mean of 6.1 ppb with a 95% confidence level of 5.1-7.5 ppb; at 30 min, the reduction was 2.8 (2.5-3.3) ppb. In children without AR, at 1 min the decline in FENO was 2.7 (2.1-3.5) ppb and by 30 min post-exercise it was 1.6 (1.3-2.0) ppb., Conclusions: The impact of exercise on FENO was dependent on the allergic phenotype, regardless of asthma status. FENO decreased immediately after exercise, and did not return to baseline level within 30 min.

Background: Nitric oxide (NO) concentration in exhaled gas is a marker of some inflammatory processes in the lung, and endogenous NO plays a role in the physiological responses to exercise and altitude. The aim of this study was to compare changes in exhaled NO concentration 5-60 mins after high-intensity exercise at 2800 m and at 180 m altitude., Methods: Twenty trained healthy volunteers (12 men), aged 19-28 years, were included in this open, crossover study. Subjects performed two exercise tests at different altitudes, 2800 m and 180 m, in a randomized order. The fraction of NO in exhaled gas (FE(NO)) was measured 5 mins before and 5-60 mins after 8 mins of running on a treadmill at a heart rate (HR) of 90% of peak HR. Peak HR was assessed during a pretest at 180 m. Ambient temperature was 20.1°C (SD = 1.2) and relative humidity 40.2% (SD = 3.2). FE(NO) measurements were corrected for altitude gas density effects and converted to partial pressure of NO (PE(NOcorr))., Results: PE(NOcorr) was reduced from 1.47 (1.21, 1.73) millipascal (mPa) at baseline to 1.11 (0.87, 1.34) mPa 5 mins after exercise at 2800 m and from 1.54 (1.24, 1.84) to 1.04 (0.87, 1.22) mPa 5 mins after exercise at 180 m. There was no difference in PE(NOcorr) between exercise at 2800 m and 180 m, and PE(NOcorr) was normalized within 20 mins., Conclusions: Exercise at 2800 m induces a similar acute reduction in exhaled nitric oxide concentration as compared with 180 m in healthy subjects., (© 2014 Scandinavian Society of Clinical Physiology and Nuclear Medicine. Published by John Wiley & Sons Ltd.)

Objective: Collateral artery growth (arteriogenesis) is an important adaptive response to hampered arterial perfusion. It is unknown whether preventive physical exercise before limb ischemia can improve arteriogenesis and modulate mononuclear cell function. This study aimed at investigating the effects of endurance exercise before arterial occlusion on MNC function and collateral artery growth., Approach and Results: After 3 weeks of voluntary treadmill exercise, ligation of the right femoral artery was performed in mice. Hindlimb perfusion immediately after surgery did not differ from sedentary mice. However, previous exercise improved perfusion restoration ≤7 days after femoral artery ligation, also when exercise was stopped at ligation. This was accompanied by an accumulation of peri-collateral macrophages and increased expression of endothelial nitric oxide synthase and inducible nitric oxide synthase (iNOS) in hindlimb collateral and in MNC of blood and spleen. Systemic monocyte and macrophage depletion by liposomal clodronate but not splenectomy attenuated exercise-induced perfusion restoration, collateral artery growth, peri-collateral macrophage accumulation, and upregulation of iNOS. iNOS-deficient mice did not show exercise-induced perfusion restoration. Transplantation of bone marrow-derived MNC from iNOS-deficient mice into wild-type animals inhibited exercise-induced collateral artery growth. In contrast to sedentary controls, thrice weekly aerobic exercise training for 6 months in humans increased peripheral blood MNC iNOS expression., Conclusions: Circulating mononuclear cell-derived inducible nitric oxide is an important mediator of exercise-induced collateral artery growth., (© 2015 American Heart Association, Inc.)

Background: Although exercise has multiple health benefits, relatively little attention has been paid to its potential therapeutic effects in those with asthma., Objective: To examine the effects of acute exercise on inflammation in physically inactive and active adults with asthma., Methods: Fourteen adults with asthma (n = 6 physically inactive, n = 8 physically active) completed (1) 30 minutes of moderate-intensity exercise on a treadmill and (2) 30 minutes of rest in random order, with 4 weeks between sessions. Exhaled nitric oxide (eNO) was measured before and after the intervention (0, 0.5, 1, 2, 4, and 24 hours). Blood inflammatory mediators were measured before and after the intervention (0, 2, and 24 hours)., Results: Physically inactive participants had a significant decrease in eNO 4 hours after exercise (-4.8 ppb, -6.4 to -0.5 ppb, P = .028), which was not observed in physically active participants (P = .362). Interluekin-1 receptor antagonist increased in the physically inactive group 2 hours after exercise, with this increase strongly correlated with the decrease in eNO at 4 hours (R = -0.685, P = .007) and 24 hours (R = -0.659, P = .014) after exercise. Interleukin-6 was increased significantly 2 hours after exercise in physically inactive participants. Blood neutrophils and nuclear factor erythroid 2-like 2 gene expression were increased 2 hours after exercise in the overall cohort., Conclusion: This study demonstrates that acute moderate-intensity exercise is associated with decreased eNO in physically inactive adults with asthma and suggests that interluekin-1 receptor antagonist could have a role in mediating this effect. The attenuated response in physically active participants might be due to the sustained anti-inflammatory effects of exercise training. Future studies should investigate the impact of exercise intensity and exercise training on airway inflammation in those with asthma., Trial Registration: Australian New Zealand Clinical Trials Registry (http://www.anzctr.org.au), registration number ACTRN12613001014741., (Copyright © 2015 American College of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.)

Objective: Previous studies have shown that estrogen deficiency, arising in postmenopause, promotes endothelial dysfunction. This study evaluated the effects of aerobic exercise training on endothelial dependent vasodilation of aorta in ovariectomized rats, specifically investigating the role of nitric oxide (NO) and reactive oxygen species (ROS)., Methods: Female Wistar rats ovariectomized (OVX - n=20) or with intact ovary (SHAM - n=20) remained sedentary (OVX and SHAM) or performed aerobic exercise training on a treadmill 5 times a week for a period of 8 weeks (OVX-TRA and SHAM-TRA). In the thoracic aorta the endothelium-dependent and -independent vasodilation was assessed by acetylcholine (ACh) and sodium nitroprusside (SNP), respectively. Certain aortic rings were incubated with L-NAME to assess the NO modulation on the ACh-induced vasodilation. The fluorescence to dihydroethidium in aortic slices and plasma nitrite/nitrate concentrations were measured to evaluate ROS and NO bioavailability, respectively., Results: ACh-induced vasodilation was reduced in OVX rats as compared SHAM (Rmax: SHAM: 86±3.3 vs. OVX: 57±3.0%, p<0.01). Training prevented this response in OVX-TRA (Rmax: OVX-TRA: 88±2.0%, p<0.01), while did not change it in SHAM-TRA (Rmax: SHAM-TRA: 80±2.2%, p<0.01). The L-NAME incubation abolished the differences in ACh-induced relaxation among groups. SNP-induced vasodilation was not different among groups. OVX reduced nitrite/nitrate plasma concentrations and increased ROS in aortic slices, training as effective to restore these parameters to the SHAM levels., Conclusions: Exercise training, even in estrogen deficiency conditions, is able to improve endothelial dependent vasodilation in rat aorta via enhanced NO bioavailability and reduced ROS levels.

The cardiovascular response to exercise is abnormally large in hypertension. Over the past decade, it has become clear that the exercise pressor reflex (a peripheral feed-back mechanism originating in skeletal muscle) contributes significantly to the generation of this hyper-responsiveness. Further, it has been determined that overactivity of the mechanically (muscle mechanoreflex) and chemically (muscle metaboreflex) sensitive components of the exercise pressor reflex underpin its dysfunction. Given the recent attention in the literature, this review focuses upon the aberrant function of the muscle mechanoreflex in this disease. Evidence supporting a role for the mechanoreflex in the pathogenesis of the exaggerated cardiovascular response to physical activity is highlighted. The peripheral and central mechanisms that may be responsible for mechanoreflex overactivity in hypertension are likewise discussed. Particular attention is given to emerging evidence implicating a role for centrally-derived nitric oxide in this process., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The fractional concentration of exhaled nitric oxide (FENO), a known marker of atopic-eosinophilic inflammation, may be used as a surrogate to assess exercise-induced bronchoconstriction (EIB) in asthmatic children. The predictive value of baseline FENO for EIB appears to be influenced by several factors, including age, atopy, current therapy with corticosteroids and measurement technique. Nonetheless, FENO cut-off values appear to be able to rule out EIB. FENO levels decrease during EIB, apparently through neural mechanisms rather than by decreased airway-epithelial surface. Partition of FENO into proximal and peripheral contributions of the respiratory tract may improve our understanding on NO exchange during exercise and help to screen subjects prone to EIB. Other biomarkers of inflammation and oxidative stress contained in exhaled gases and exhaled breath condensate (EBC) may shed light on the pathophysiology of EIB. Exhaled breath temperature is a promising real-time measurement whose routine use for assessing EIB warrants further investigation., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Nitric oxide is produced within skeletal muscle fibres and has various functions in skeletal muscle. There is evidence that NO may be essential for normal increases in skeletal muscle glucose uptake during contraction/exercise. Although there have been some discrepant results, it has been consistently demonstrated that inhibition of NO synthase (NOS) attenuates the increase in skeletal muscle glucose uptake during contraction in mouse and rat muscle ex vivo, during in situ contraction in rats and during exercise in humans. The NO-mediated increase in skeletal muscle glucose uptake during contraction/exercise is probably due to the modulation of intramuscular signalling that ultimately increases glucose transporter 4 (GLUT4) translocation and is, surprisingly, independent of blood flow. In this review, we discuss the evidence for and against a role of NO in regulating skeletal muscle glucose uptake during contraction/exercise and outline the possible mechanism(s) involved. Emerging findings regarding the role of neuronal NOS mu (nNOSμ) in this process are also discussed., (© 2014 The Authors. Experimental Physiology © 2014 The Physiological Society.)

Background: The mechanisms of smoking tobacco leading to chronic obstructive pulmonary disease (COPD) are beginning to be understood. However, conclusions about the role of blood or lung oxidative stress markers were disparate., Aims: To investigate the oxidative stress in blood or lung associated with tobacco smoke and to evaluate its effect on pulmonary function data and its relation with physical activity., Methods: It is a case-control study. Fifty-four male-smokers of more than five pack-years (PY) and aged 40-60 years were included (29 Non-COPD, 16 COPD). Physical activity score was determined. Blood sample levels of malondialdehyde (MDA), protein-cys-SH (PSH), and Glutathione (GSH) were measured. Fractional exhaled nitric oxide (FeNO) and plethysmographic measurements were performed. Correlation coefficients (r) evaluated the association between oxidative stress markers and independent variables (plethysmographic data and physical activity score)., Results: Non-COPD (48 ± 6 years) and COPD (49 ± 5 years) groups had similar tobacco consumption patterns, that is, 27 ± 14 PY versus 30 ± 19 PY, respectively. Compared to the Non-COPD group, the COPD group had significantly lower levels of GSH and PSH, that is, mean ± SE were 40 ± 6 versus 25 ± 5 µg/mL and 54 ± 10 versus 26 ± 5 µg/g of hemoglobin, respectively. However, MDA level and FeNO values were similar. In the COPD group, none of the oxidative stress markers was significantly correlated with plethysmographic data or physical activity score. In the Non-COPD group, GSH was significantly correlated with physical activity score (r = 0.47) and PSH was significantly correlated with total lung capacity (TLC) (r = -0.50), residual volume (r = 0.41), and physical activity score (r = 0.62). FeNO was significantly correlated with TLC of the COPD group (r = -0.48)., Conclusion: Compared to the Non-COPD group, the COPD group had a marked decrease in blood antioxidant markers (GSH and PSH) but similar blood oxidant (MDA) or lung (FeNO) burden.

Nitric oxide (NO) is a signalling molecule that contributes to the control of many physiological pathways, including the heat-loss response of skin vasodilatation. Recently, NO has been implicated in the control of sweating during exercise in young adults. We tested the hypothesis that ageing reduces NO-dependent sweating during exercise in the heat. Ten young (23 ± 3 years old) and 10 older men (64 ± 5 years old), matched for body surface area, performed three successive 15 min bouts of exercise (Ex1, Ex2 and Ex3) at the same rate of metabolic heat production (300 W m(-2)) in the heat (35°C, 20% relative humidity). Exercise periods were interspersed with 15 min recovery periods. Local sweat rate (ventilated capsule) was measured on two forearm skin sites, which were continuously perfused via intradermal microdialysis with 0.9% saline as control (CON) or 10 mm N(G)-nitro-l-arginine methyl ester (L-NAME), a non-selective NO synthase inhibitor. Local sweat rate at the end of Ex1 was lower in the CON conditions in the older versus young men (0.69 ± 0.19 versus 0.90 ± 0.17 mg min(-1) cm(-2), P = 0.018). In the young men, local sweat rate was reduced in the L-NAME-treated conditions compared with the CON conditions at the end of Ex1 (0.67 ± 0.14 versus 0.90 ± 0.17 mg min(-1) cm(-2), P = 0.004), Ex2 (0.78 ± 0.20 versus 1.03 ± 0.20 mg min(-1) cm(-2), P = 0.013) and Ex3 (0.78 ± 0.20 versus 1.03 ± 0.21 mg min(-1) cm(-2), P = 0.014). In the older men, there was no main effect of treatment conditions on local sweat rate (P = 0.537) such that local sweat rates in the L-NAME-treated and CON conditions were similar (Ex1, 0.65 ± 0.20 versus 0.69 ± 0.19 mg min(-1) cm(-2); Ex2, 0.80 ± 0.27 versus 0.91 ± 0.29 mg min(-1) cm(-2); and Ex3, 0.84 ± 0.31 versus 0.94 ± 0.38 mg min(-1) cm(-2)). We conclude that ageing attenuates the influence of NO in the control of local forearm sweating observed in young adults during short 15 min bouts of exercise in the heat. This mechanism may, in part, explain the age-related impairments in sweating., (© 2014 The Authors. Experimental Physiology © 2014 The Physiological Society.)

Type 2 diabetes mellitus associated with obesity, or "diabesity", coincides with an altered nitric oxide (NO) metabolism in skeletal muscle. Three isoforms of nitric oxide synthase (NOS) exist in human skeletal muscle tissue. Both neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS) are constitutively expressed under physiological conditions, producing low levels of NO, while the inducible nitric oxide synthase (iNOS) is strongly up-regulated only under pathophysiological conditions, excessively increasing NO concentrations. Due to chronic inflammation, overweight/obese type 2 diabetic patients exhibit up-regulated protein contents of iNOS and concomitant elevated amounts of NO in skeletal muscle. Low muscular NO levels are important for attaining an adequate cellular redox state--thereby maintaining metabolic integrity--while high NO levels are believed to destroy cellular components and to disturb metabolic processes, e.g., through strongly augmented posttranslational protein S-nitrosylation. Physical training with submaximal intensity has been shown to attenuate inflammatory profiles and iNOS protein contents in the long term. The present review summarizes signaling pathways which induce iNOS up-regulation under pathophysiological conditions and describes molecular mechanisms by which high NO concentrations are likely to contribute to triggering skeletal muscle insulin resistance and to reducing mitochondrial capacity during the development and progression of type 2 diabetes. Based on this information, it discusses the beneficial effects of regular physical exercise on the altered NO metabolism in the skeletal muscle of overweight/obese type 2 diabetic subjects, thus unearthing new perspectives on training strategies for this particular patient group., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Aims: To investigate the effect of 16 weeks of aerobic training performed at two different intensities on nitric oxide (tNOx) availability and iNOS/nNOS expression, oxidative stress (OS) and inflammation in obese humans with or without type 2 diabetes mellitus (T2DM)., Methods: Twenty-five sedentary, obese (BMI > 30 kg/m2) males (52.8 ± 7.2 years); 12 controls versus 13 T2DM were randomly allocated to four groups that exercised for 30 min, three times per week either at low (Fat-Max; 30-40% VO(2max)) or moderate (T(vent); 55-65 % VO(2max)) intensity. Before and after training, blood and muscle samples (v. lateralis) were collected., Results: Baseline erythrocyte glutathione was lower (21.8 ± 2.8 vs. 32.7 ± 4.4 nmol/ml) and plasma protein oxidative damage and IL-6 were higher in T2DM (141.7 ± 52.1 vs. 75.5 ± 41.6 nmol/ml). Plasma catalase increased in T2DM after T(vent) training (from 0.98 ± 0.22 to 1.96 ± 0.3 nmol/min/ml). T2DM groups demonstrated evidence of oxidative damage in response to training (elevated protein carbonyls). Baseline serum tNOx were higher in controls than T2DM (18.68 ± 2.78 vs. 12.34 ± 3.56 μmol/l). Training at T(vent) increased muscle nNOS and tNOx in the control group only. Pre-training muscle nNOS was higher in controls than in T2DMs, while the opposite was found for iNOS. No differences were found after training for plasma inflammatory markers., Conclusion: Exercise training did not change body composition or aerobic fitness, but improved OS markers, especially when performed at T(vent). Non-diabetics responded to T(vent) training by increasing muscle nNOS expression and tNOx levels in skeletal muscle while these parameters did not change in T2DM, perhaps due to higher insulin resistance (unchanged after intervention).

Nitric oxide (NO) is known to be one of the most important regulatory compounds within the cardiovascular system where it is central for functions such as regulation of blood pressure, blood flow, and vascular growth. The bioavailability of NO is determined by a balance between, on one hand, the extent of enzymatic and non-enzymatic formation of NO and on the other hand, removal of NO, which in part is dependent on the reaction of NO with reactive oxygen species (ROS). The presence of ROS is dependent on the extent of ROS formation via mitochondria and/or enzymes such as NAD(P)H oxidase (NOX) and xanthine oxidase (XO) and the degree of ROS removal through the antioxidant defense system or other reactions. The development of cardiovascular disease has been proposed to be closely related to a reduced bioavailability of NO in parallel with an increased presence of ROS. Excessive levels of ROS not only lower the bioavailability of NO but may also cause cellular damage in the cardiovascular system. Physical activity has been shown to greatly improve cardiovascular function, in part through improved bioavailability of NO, enhanced endogenous antioxidant defense and a lowering of the expression of ROS-forming enzymes. Regular physical activity is therefore likely to be a highly useful tool in the treatment of cardiovascular disease. Future studies should focus on which form of exercise may be most optimal for enhancing NO bioavailability and improving cardiovascular health.

Aims: Eighteen patients with asthma were evaluated during preparation to climb to extreme altitude in order to study the effects of low fractional inspired oxygen (FiO(2)), prolonged exposure to cold air and high altitude on lung function, asthma control and airway inflammation., Methods: Spirometry and airway inflammation (fractional exhaled nitric oxide (FeNO) and induced sputum) were studied under different test conditions: hypoxic (FiO(2)=11%) exercise test, 24-hour cold exposure (-5°C) and before, during and after an expedition that involved climbing the Aconcagua mountain (6965 m)., Results: Forced expiratory volume in 1 s (FEV(1)) and FeNO values were slightly lower (p<0.04) after 1 h of normobaric hypoxia. FEV(1) decreased (p=0.009) after 24-hour cold exposure, accompanied by increased sputum neutrophilia (p<0.01). During the expedition FEV(1) and forced vital capacity decreased (maximum FEV(1) decrease of 12.3% at 4300 m) and asthma symptoms slightly increased. After the expedition the Asthma Control Test score and prebronchodilator FEV(1) were reduced (p<0.02), sputum neutrophil count was increased (p=0.04) and sputum myeloperoxidase levels, sputum interleukin 17 mRNA, serum and sputum vascular endothelial growth factor A levels were significantly higher compared with baseline. Patients with asthma with the lowest oxygen saturation during the hypoxic exercise test were more prone to develop acute mountain sickness., Conclusions: Exposure to environmental conditions at high altitude (hypoxia, exercise, cold) was associated with a moderate loss of asthma control, increased airway obstruction and neutrophilic airway inflammation. The cold temperature is probably the most important contributing factor as 24-hour cold exposure by itself induced similar effects.

Introduction: Exhaled nitric oxide (FENO) measurements are recommended to be performed before spirometry and exercise challenge tests because forced breathing might influence FENO values. Information on the effect of exercise on FENO is lacking in non-asthmatic children., Aim:   To investigate the effect on FENO of a standardized exercise challenge test on a treadmill in non-asthmatic children with and without allergic rhinoconjunctivitis (AR) symptoms., Methods: From the case-control study 'Asthma and allergy among school children in Nordland', 330 non-asthmatic pupils age 8-16 years were enrolled. FENO was measured at baseline and at 1 min and 30 min after exercise challenge test by the single breath technique with EcoMedics Exhalazer® (Eco Physics, Duernten, Switzerland)., Results: Pair-wise comparison of FENO from baseline demonstrated a highly significant reduction in FENO post-exercise for all children at 1 min (27.4%) and at 30 min (16.1%) (P < 0.001). The AR group had a significantly higher decline in FENO value at 1 min post-exercise compared to the non-AR group, 4.2 parts per billion (ppb) vs 2.6 ppb (P < 0.001). Decline in FENO immediately post-exercise was more significant if baseline FENO was ≥ 20 ppb; mean reduction 9.9 (95% CI: 8.7-11.4) ppb., Conclusion: FENO is reduced by 27.4% immediately after a standardized treadmill exercise test in non-asthmatic children. Pupils reporting AR symptoms demonstrate a larger decline in FENO value at 1 min post-exercise compared to pupils without AR symptoms. These findings confirm that children should refrain from physical activity before FENO measurement., (© 2012 Blackwell Publishing Ltd.)

Background and Aims: Whether intensity or other characteristics of physical activity can better promote the release of nitric oxide (NO) and reduction of blood pressure in hypertensive older-adults is still unknown. In this study, the post-exercise blood pressure (BP) response and NO release after different intensities of aerobic exercise in elderly women were analyzed., Methods: Blood pressure response and NO were analyzed in 23 elderly mildly hypertensive women. Participants underwent (1) high-intensity incremental exercise (IT); (2) moderate-intensity 20 min exercise at 90% of the anaerobic threshold (AT), and (3) control (CONT) session. BP was measured before and after interventions; volunteers remained seated for 1 h. NO estimates were made through NO2- analyses., Results: After CONT session, both diastolic BP and mean arterial pressure (MAP) were significantly higher than during pre-exercise resting. Post-exercise hypotension (PEH) was observed after exercise at IT and 90% of AT. Although exercise in both sessions lowered SBP and MAP compared with CONT, exercise at the highest intensity (IT) was more effective on lowering systolic BP after exercise. In comparison with pre-exercise resting, NO2- increased significantly only after IT, but both exercise sessions caused NO2- to increase compared with CONT., Conclusion: Exercise intensity and NO release may exert a role in eliciting PEH in mildly hypertensive elderly women.

Skeletal muscle is responsible for altered acute and chronic workload as induced by exercise. Skeletal muscle adaptations range from immediate change of contractility to structural adaptation to adjust the demanded performance capacities. These processes are regulated by mechanically and metabolically induced signaling pathways, which are more or less involved in all of these regulations. Nitric oxide is one of the central signaling molecules involved in functional and structural adaption in different cell types. It is mainly produced by nitric oxide synthases (NOS) and by non-enzymatic pathways also in skeletal muscle. The relevance of a NOS-dependent NO signaling in skeletal muscle is underlined by the differential subcellular expression of NOS1, NOS2, and NOS3, and the alteration of NO production provoked by changes of workload. In skeletal muscle, a variety of highly relevant tasks to maintain skeletal muscle integrity and proper signaling mechanisms during adaptation processes towards mechanical and metabolic stimulations are taken over by NO signaling. The NO signaling can be mediated by cGMP-dependent and -independent signaling, such as S-nitrosylation-dependent modulation of effector molecules involved in contractile and metabolic adaptation to exercise. In this review, we describe the most recent findings of NO signaling in skeletal muscle with a special emphasis on exercise conditions. However, to gain a more detailed understanding of the complex role of NO signaling for functional adaptation of skeletal muscle (during exercise), additional sophisticated studies are needed to provide deeper insights into NO-mediated signaling and the role of non-enzymatic-derived NO in skeletal muscle physiology.

Polycystic ovary syndrome (PCOS) is associated with cardiovascular disease. The contribution of the nitric oxide (NO) dilator system to cutaneous endothelial dysfunction is currently unknown in PCOS. Our aim was to examine whether women with PCOS demonstrate impaired cutaneous microvascular NO function and whether exercise training can ameliorate any impairment. Eleven women with PCOS (age, 29 ± 7 years; body mass index, 34 ± 6 kg m(-2)) were compared with six healthy obese control women (age, 29 ± 7 years; body mass index, 34 ± 5 kg m(-2)). Six women with PCOS (30 ± 7 years; 31 ± 6 kg m(-2)) then completed 16 weeks of exercise training. Laser Doppler flowmetry, combined with intradermal microdialysis of l-N(G)-monomethyl-l-arginine, a nitric oxide antagonist, in response to incremental local heating of the forearm was assessed in women with PCOS and control women, and again in women with PCOS following exercise training. Cardiorespiratory fitness, homeostasis model assessment for insulin resistance, hormone and lipid profiles were also assessed. Differences between women with PCOS and control women and changes with exercise were analysed using Student's unpaired t tests. Differences in the contribution of NO to cutaneous blood flow [expressed as a percentage of maximal cutaneous vasodilatation (CVCmax)] were analysed using general linear models. At 42°C heating, cutaneous NO-mediated vasodilatation was attenuated by 17.5%CVCmax (95% confidence interval, 33.3, 1.7; P = 0.03) in women with PCOS vs. control women. Exercise training improved cardiorespiratory fitness by 5.0 ml kg(-1) min(-1) (95% confidence interval, 0.9, 9.2; P = 0.03) and NO-mediated cutaneous vasodilatation at 42°C heating by 19.6% CVCmax (95% confidence interval, 4.3, 34.9; P = 0.02). Cutaneous microvascular NO function is impaired in women with PCOS compared with obese matched control women but can be improved with exercise training.

We tested the hypothesis that nitric oxide (NO) and adenosine contribute to the onset of vasodilation during dynamic forearm exercise. Twenty-two subjects performed rhythmic forearm exercise (20 % of maximum) during control and NO synthase (NOS) inhibition (N (G)-monomethyl-L-arginine; L-NMMA) trials. A subset of subjects performed a third trial of forearm exercise during combined inhibition of NOS and adenosine (aminophylline; n = 9). Additionally, a separate group of subjects (n = 7) performed rhythmic forearm exercise during control, inhibition of adenosine alone and combined inhibition of adenosine and NOS. Forearm vascular conductance (FVC; ml min(-1) · 100 mmHg(-1)) was calculated from blood flow and mean arterial pressure (mmHg). The onset of vasodilation was assessed by calculating the slope of the FVC response for every duty cycle between baseline and steady state, and the number of duty cycles (1-s contraction/2-s relaxation) to reach steady state. NOS inhibition blunted vasodilation at the onset of exercise (11.1 ± 0.8 vs. 8.5 ± 0.6 FVC units/duty cycle; P < 0.001 vs. control) and increased the time to reach steady state (25 ± 1 vs. 32 ± 1 duty cycles; P < 0.001 vs. control). Vasodilation was blunted further with combined inhibition of NOS and adenosine (7.5 ± 0.6 vs. 6.2 ± 0.8 FVC units/duty cycle; P < 0.05 vs. L-NMMA alone), but not with aminophylline alone (16.0 ± 2.2 vs. 14.7 ± 2.0 FVC units/duty cycle; P = 0.67 vs. control). Our data indicate that NO and adenosine (in the absence of NO) contribute to the onset of vasodilation during dynamic forearm exercise.

Purpose: Slight differences in physiological responses and nitric oxide (NO) have been reported at rest between hypobaric hypoxia (HH) and normobaric hypoxia (NH) during short exposure.Our study reports NO and oxidative stress at rest and physiological responses during moderate exercise in HH versus NH., Methods: Ten subjects were randomly exposed for 24 h to HH (3000 m; FIO2, 20.9%; BP, 530 ± 6 mm Hg) or to NH (FIO2, 14.7%; BP, 720 ± 1 mm Hg). Before and every 8 h during the hypoxic exposures, pulse oxygen saturation (SpO2), HR, and gas exchanges were measured during a 6-min submaximal cycling exercise. At rest, the partial pressure of exhaled NO, blood nitrate and nitrite (NOx), plasma levels of oxidative stress, and pH levels were additionally measured., Results: During exercise, minute ventilation was lower in HH compared with NH (-13% after 8 h, P < 0.05). End-tidal CO2 pressure was lower (P < 0.01) than PRE both in HH and NH but decreased less in HH than that in NH (-25% vs. -37%, P < 0.05).At rest, exhaled NO and NOx decreased in HH (-46% and -36% after 24 h, respectively, P < 0.05) whereas stable in NH. By contrast, oxidative stress was higher in HH than that in NH after 24 h (P < 0.05). The plasma pH level was stable in HH but increased in NH (P < 0.01). When compared with prenormoxic values, SpO2, HR, oxygen consumption, breathing frequency, and end-tidal O2 pressure showed similar changes in HH and NH., Conclusion: Lower ventilatory responses to a similar hypoxic stimulus during rest and exercise in HH versus NH were sustained for 24 h and associated with lower plasma pH level, exaggerated oxidative stress, and impaired NO bioavailability.

We tested whether the known cytochrome c oxidase (COX) inhibition by nitric oxide (NO) could be quantified by VO(2) kinetics during constant load supra-Anaerobic Threshold (AT) exercises in healthy trained or untrained subjects following aerobic training or nitrate administration. In cycle ergometer constant load exercises supra-AT, identified in previous incremental tests, VO(2) kinetics describe a double exponential curve, one rapid and one appreciably slower, allowing the area between them to be calculate in O(2) l. After training, with increased NO availability, this area decreases in inverse ratio to treatment efficacy. In fact, in 11 healthy subjects after aerobic training for 6-7 weeks, area was decreased on average by 51 %. In 11 untrained subjects, following the assumption of an NO donor, 20 mg isosorbide 5 mononitrate, area was decreased on average by 53 %. In conclusion, supra-AT VO(2) kinetics in constant load exercises permit the quantification of the inhibitory effect NO-dependent on COX after either physical training or nitrate assumption.

Background: Fractional exhaled nitric oxide (eNO) is recognized as a marker of pulmonary endothelial function. Oxidative stress is associated with systemic endothelial nitric oxide production, but its correlation with eNO in heart failure (HF) patients has not been described. Previous studies have reported increased eNO levels after exercise in symptomatic HF patients but decreased levels with pulmonary arterial hypertension. Our objective was to prospectively examine the potential myocardial and functional determinants of exercise-induced rise of eNO in HF., Methods and Results: Thirty-four consecutive ambulatory patients with chronic systolic HF (left ventricular ejection fraction [LVEF] ≤45%) underwent symptom-limited cardiopulmonary stress testing and echocardiography. eNO was determined immediately after exercise. Systemic endothelial dysfunction was assessed by asymmetric dimethylarginine (ADMA) and the L-arginine/ADMA ratio. In our study cohort (mean age 53 ± 13 years, 76% male, median LVEF 31%, interquartile range [IQR] 25%-40%), the mean eNO was 23 ± 9 ppb. eNO levels were higher in patients with diastolic dysfunction stages 2 or 3 than stage 1 or normal diastology (26.1 ± 9 vs 19.5 ± 7 ppb; P = .013). eNO had a positive correlation with estimated systolic pulmonary artery pressure (r = 0.57; P = .0009) and indexed left atrium volume (r = 0.43; P = .014), but it did not correlate with cardiopulmonary exercise test parameters, ADMA, or symptom score., Conclusions: In contrast to earlier reports, the increase in postexercise eNO observed in stable chronic systolic HF patients may be attributed to the presence of underlying pulmonary venous hypertension probably secondary to advanced diastolic dysfunction., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Nitric oxide (NO) is an important vasodilator and regulator in the cardiovascular system, and this link was the subject of a Nobel prize in 1998. However, NO also plays many other regulatory roles, including thrombosis, immune function, neural activity, and gastrointestinal function. Low concentrations of NO are thought to have important signaling effects. In contrast, high concentrations of NO can interact with reactive oxygen species, causing damage to cells and cellular components. A less-recognized site of NO production is within skeletal muscle, where small increases are thought to have beneficial effects such as regulating glucose uptake and possibly blood flow, but higher levels of production are thought to lead to deleterious effects such as an association with insulin resistance. This review will discuss the role of NO in skeletal muscle during and following exercise, including in mitochondrial biogenesis, muscle efficiency, and blood flow with a particular focus on its potential role in regulating skeletal muscle glucose uptake during exercise.

The roles of local metabolites in reactive and exercise hyperemia remain incompletely understood. A maximum metabolic stimulus caused by ischemic exercise (IE) could potentially fully activate all vasodilator pathways and limit potential redundancy amongst vasoactive substances. We tested the hypotheses that IE elicits a reproducible hyperemic response in the forearm and that adenosine (ADO) and nitric oxide (NO) contribute to this response. In separate protocols, forearm blood flow (FBF) was measured with venous occlusion plethysmography following IE trials consisting of 5 min of ischemia and rhythmic forearm handgrip exercise (performed during last 2 min of ischemia). In protocol 1 (n = 8), FBF was measured after three trials of IE. In protocol 2 (n = 9), subjects performed IE during control (saline), aminophylline (APH; adenosine receptor antagonist), and combined APH/N (G)-monomethyl-L-arginine (L-NMMA; NOS inhibition) infusions. In protocol 1, coefficients of variation for total (area under the curve) ΔFBF and ΔFVC (forearm vascular conductance) following IE were 10.4 ± 1.0% and 14.9 ± 1.0%, respectively. In protocol 2, peak ΔFBF was similar for saline and APH trials. Peak ΔFBF for the APH+L: -NMMA trial was greater than that of the APH trial (P = 0.03), and peak ΔFVC was marginally non-significant (P = 0.053). Total ΔFBF (54.8 ± 3.9, 55.2 ± 5.4, and 60.4 ± 4.8 ml 100 ml(-1); P = 0.43) and ΔFVC (51.4 ± 3.5, 52.1 ± 5.5, and 56.5 ± 5.0 ml 100 ml(-1) 100 mmHg(-1); P = 0.52) were similar for saline, APH, and APH+L: -NMMA, respectively. Our data suggest that (1) the hyperemic response to IE is reproducible and (2) inhibition of ADO alone or combined ADO and NO does not blunt the hyperemic response following IE.

Physical exercise mobilizes endothelial progenitor cells (EPCs) to peripheral blood. However, this effect seems to depend on exercise characteristics, such as duration and intensity. The aim of this systematic review was to verify the impact of a single bout of aerobic exercise on the mobilization of EPCs in healthy individuals, and the potential mechanisms involved. The bibliographic search was conducted on the following electronic databases in May 2011: SciELO, LILACS, Cochrane, ClinicalTrials.gov, SPORTDiscus and Medline. Of the 178 articles initially identified, 12 met the inclusion criteria and were classified regarding quality according to the PEDro scale. The magnitude and duration of the EPC mobilization response were higher after long/ultralong duration exercises, and they are correlated with vascular endothelial growth factor (VEGF) plasma levels. The EPC mobilization peak in response to a maximal or submaximal single bout of exercise lasting up to one hour occurs immediately after the exercise or within the first hour after it. One possible mechanism is nitric oxide (NO) bioavailability. The individuals' age and exercise intensity seem to interfere with the EPC mobilization response. Long/ultralong duration exercises promote more pronounced EPC mobilization as compared with maximal or submaximal exercises. The mechanisms involve VEGF release in long/ultralong duration exercises and NO bioavailability in maximal or submaximal exercises lasting less than one hour.

Recent studies have investigated the influence of dietary nitrate supplementation on the physiological responses to exercise. Specifically, it has been reported that enhancing nitric oxide (NO) bioavailability through supplementation of the diet with nitrate salts or nitrate-rich beetroot juice reduces the O(2) cost of exercise and improves exercise performance. The lower O(2) cost for a given sub-maximal work rate following nitrate ingestion indicates that muscle efficiency is enhanced either as a consequence of a reduced energy cost of contraction or enhanced mitochondrial efficiency. The positive effects of nitrate supplementation on the O(2) cost of sub-maximal exercise can be manifested acutely (i.e. 2.5 h following ingestion) and maintained for at least 15 days if supplementation is continued. Most recently, the influence of dietary nitrate supplementation on time trial performance in competitive cyclists has been investigated. Studies have shown a 1-2% reduction in the time to complete time trial distances between 4 and 16 km. The dose of nitrate that has been shown to improve exercise efficiency can readily be achieved through the consumption of 0.5 litre of beetroot juice or an equivalent high-nitrate foodstuff. Following a 5- to 6-mmol bolus of nitrate, plasma [nitrite] typically peaks within 2-3 h and remains elevated for a further 6-9 h before declining towards baseline. Therefore, consuming nitrate approximately 3 h prior to competition or training is recommended if athletes wish to explore the ergogenic potential of nitrate supplementation., (Copyright © 2012 S. Karger AG, Basel.)

Background: The absence of the I allele of the angiotensin converting enzyme (ACE) gene has been associated with higher levels of circulating ACE, lower nitric oxide (NO) release and hypertension. The purposes of this study were to analyze the post-exercise salivary nitrite (NO2-) and blood pressure (BP) responses to different exercise intensities in elderly women divided according to their ACE genotype., Methods: Participants (n = 30; II/ID = 20 and DD = 10) underwent three experimental sessions: incremental test - IT (15 watts workload increase/3 min) until exhaustion; 20 min exercise 90% anaerobic threshold (90% AT); and 20 min control session without exercise. Volunteers had their BP and NO2- measured before and after experimental sessions., Results: Despite both intensities showed protective effect on preventing the increase of BP during post-exercise recovery compared to control, post-exercise hypotension and increased NO2- release was observed only for carriers of the I allele (p < 0.05)., Conclusion: Genotypes of the ACE gene may exert a role in post-exercise NO release and BP response.

Exercise hyperaemia in hypoxia is augmented relative to the same level of exercise in normoxia. At moderate exercise intensities, the mechanism(s) underlying this augmented response are currently unclear. We tested the hypothesis that endothelium-derived nitric oxide (NO) and vasodilating prostaglandins (PGs) contribute to the augmented muscle blood flow during hypoxic exercise relative to normoxia. In 10 young healthy adults, we measured forearm blood flow (FBF; Doppler ultrasound) and calculated the vascular conductance (FVC) responses during 5 min of rhythmic handgrip exercise at 20% maximal voluntary contraction in normoxia (NormEx) and isocapnic hypoxia (HypEx; O2 saturation ∼85%) before and after local intra-brachial combined blockade of NO synthase (NOS; via N(G)-monomethyl-L-arginine: L-NMMA) and cyclooxygenase (COX; via ketorolac). All trials were performed during local α- and β-adrenoceptor blockade to eliminate sympathoadrenal influences on vascular tone and thus isolate local vasodilatation. Arterial and deep venous blood gases were measured and oxygen consumption (VO2) was calculated. In control (saline) conditions, FBF after 5 min of exercise in hypoxia was greater than in normoxia (345 ± 21 ml min(−1) vs. 297 ± 18 ml min(−1); P < 0.05). After NO–PG block, the compensatory increase in FBF during hypoxic exercise was blunted ∼50% and thus was reduced compared with control hypoxic exercise (312 ± 19 ml min(−1); P < 0.05), but this was not the case in normoxia (289 ± 15 ml min(−1); P = 0.33). The lower FBF during hypoxic exercise was associated with a compensatory increase in O2 extraction, and thus VO2 was maintained at normal control levels (P = 0.64–0.99). We conclude that under the experimental conditions employed, NO and PGs have little role in normoxic exercise hyperaemia whereas combined NO–PG inhibition reduces hypoxic exercise hyperaemia and abolishes hypoxic vasodilatation at rest. Additionally, VO2 of the tissue was maintained in hypoxic conditions at rest and during exercise, despite attenuated oxygen delivery following NO–PG blockade, due to an increase in O2 extraction at the level of the muscle.

Rationale: Exercise training confers sustainable protection against ischemia-reperfusion injury in animal models and has been associated with improved survival following a heart attack in humans. It is still unclear how exercise protects the heart, but it is apparent that endothelial nitric oxide synthase (eNOS) and nitric oxide (NO) play a role., Objective: To determine the role of β(3)-adrenergic receptors (β(3)-ARs), eNOS activation, and NO metabolites (nitrite and nitrosothiols) in the sustained cardioprotective effects of exercise., Methods and Results: Here we show that voluntary exercise reduces myocardial injury in mice following a 4-week training period and that these protective effects can be sustained for at least 1 week following the cessation of the training. The sustained cardioprotective effects of exercise are mediated by alterations in the phosphorylation status of eNOS (increase in serine 1177 and decrease in threonine 495), leading to an increase in NO generation and storage of NO metabolites (nitrite and nitrosothiols) in the heart. Further evidence revealed that the alterations in eNOS phosphorylation status and NO generation were mediated by β(3)-AR stimulation and that in response to exercise a deficiency of β(3)-ARs leads to an exacerbation of myocardial infarction following ischemia-reperfusion injury., Conclusions: Our findings clearly demonstrate that exercise protects the heart against myocardial ischemia-reperfusion injury by stimulation of β(3)-ARs and increased cardiac storage of nitric oxide metabolites (ie, nitrite and nitrosothiols).