Assessing exogenous carbohydrate intake needed to optimize human endurance performance across sex: insights from modeling runners pursuing a sub-2-h marathon.

Carbohydrate (CHO) availability sustains high metabolic demands during prolonged exercise. The adequacy of current CHO intake recommendations, 30-90 g·h-1 dependent on CHO mixture and tolerability, to support elite marathon performance is unclear. We sought to scrutinize the current upper limit recommendation for exogenous CHO intake to support modeled sub-2-h marathon (S2M) attempts across elite male and female runners. Male and female runners (n = 120 each) were modeled from published literature with reference characteristics necessary to complete a S2M (e.g., body mass and running economy). Completion of a S2M was considered across a range of respiratory exchange rates, with maximal starting skeletal muscle and liver glycogen content predicted for elite male and female runners. Modeled exogenous CHO bioavailability needed for male and female runners were 93 ± 26 and 108 ± 22 g·h-1, respectively (P < 0.0001, d = 0.61). Without exogenous CHO, males were modeled to deplete glycogen in 84 ± 7 min, females in 71 ± 5 min (P < 0.0001, d = 2.21) despite higher estimated CHO oxidation rates in males (5.1 ± 0.5 g·h-1) than females (4.4 ± 0.5 g·h-1; P < 0.0001, d = 1.47). Exogenous CHO intakes ≤ 90 g·h-1 are insufficient for 65% of modeled runners attempting a S2M. Current recommendations to support marathon performance appear inadequate for elite marathon runners but may be more suitable for male runners in pursuit of a S2M (56 of 120) than female runners (28 of 120).NEW & NOTEWORTHY This study scrutinizes the upper limit of exogenous carbohydrate (CHO) recommendations for elite male and female marathoners by modeling sex-specific needs across an extreme metabolic challenge lasting ∼2 h, a sub-2-h marathon. Contemporary nutritional guidelines to optimize marathon performance appear inadequate for most elite marathon runners but appear more appropriate for males over their female counterparts. Future research examining possible benefits of exogenous CHO intakes > 90 g·h-1 should prioritize female athlete study inclusion.

Muscle oxygen saturation rates coincide with lactate-based exercise thresholds.

INTRODUCTION: Monitoring muscle metabolic activity via blood lactate is a useful tool for understanding the physiological response to a given exercise intensity. Recent indications suggest that skeletal muscle oxygen saturation (SmO2), an index of the balance between local O2 supply and demand, may describe and predict endurance performance outcomes. PURPOSE: We tested the hypothesis that SmO2 rate is tightly related to blood lactate concentration across exercise intensities, and that deflections in SmO2 rate would coincide with established blood lactate thresholds (i.e., lactate thresholds 1 and 2). METHODS: Ten elite male soccer players completed an incremental running protocol to exhaustion using 3-min work to 30 s rest intervals. Blood lactate samples were collected during rest and SmO2 was collected continuously via near-infrared spectroscopy from the right and left vastus lateralis, left biceps femoris and the left gastrocnemius. RESULTS: Muscle O2 saturation rate (%/min) was quantified after the initial 60 s of each 3-min segment. The SmO2 rate was significantly correlated with blood lactate concentrations for all muscle sites; RVL, r = - 0.974; LVL, r = - 0.969; LG, r = - 0.942; LHAM, r = - 0.907. Breakpoints in SmO2 rate were not significantly different from LT1 or LT2 at any muscle sites (P > 0.05). Bland-Altman analysis showed speed threshold estimates via SmO2 rate and lactate are similar at LT2, but slightly greater for SmO2 rate at LT1. CONCLUSIONS: Muscle O2 saturation rate appears to provide actionable information about maximal metabolic steady state and is consistent with bioenergetic reliance on oxygen and its involvement in the attainment of metabolic steady state.

Identification of maximal steady-state metabolic rate by the change in muscle oxygen saturation.

We tested the hypothesis that a %SmO2 (muscle O2 saturation) slope can distinguish the heavy-severe exercise domain boundary and the highest steady-state metabolic rate. Thirteen participants (5 women) performed a graded exercise test (GXT) to determine peak oxygen consumption (V̇o2peak) and lactate turn point (LTP). On a separate study day, a %SmO2 zero-slope prediction trial included completing 5-min cycling bouts in an estimated heavy domain, at an estimated critical power, and in an estimated severe domain. Linear regression then determined the work rate at the predicted %SmO2 zero-slope, before a fourth 5-min confirmation trial. Two separate validation study days included confirmed steady-state (heavy domain) and nonsteady-state (severe domain) constant work rate trials. The power at the predicted %SmO2 zero-slope was 204 ± 36 W and occurred at a %SmO2 slope of 0.7 ± 1.4%/min (P = 0.12 relative to zero). There was no difference between the power at LTP (via GXT) and the predicted %SmO2 zero-slope linked power (P = 0.74). From validation study days, the %SmO2 slope was 0.32 ± 0.73%/min during confirmed heavy-domain constant work rate exercise and -0.75 ± 1.94%/min during confirmed severe-domain exercise (P < 0.05). The %SmO2 zero-slope consistently delineated steady state from nonsteady-state metabolic parameters (V̇o2 and blood lactate) and the heavy-severe domain boundary. Our data suggest the %SmO2 slope can identify the highest steady-state metabolic rate and the physiological boundary between the heavy-severe domain, independent of work rate.NEW & NOTEWORTHY Muscle O2 saturation (%SmO2) rate can be used to not only identify sustainable from unsustainable exercise intensities but also delineate the transition from heavy to severe exercise domains. This report is the first to identify, and then validate, that the highest steady-state metabolic rate is related to a zero-slope muscle O2 saturation and is therefore dependent on muscle oxygen supply-demand balance.

Highly Cushioned Shoes Improve Running Performance in Both the Absence and Presence of Muscle Damage.

PURPOSE: We tested the hypotheses that a highly cushioned running shoe (HCS) would 1) improve incremental exercise performance and reduce the oxygen cost (Oc) of submaximal running, and 2) attenuate the deterioration in Oc elicited by muscle damage consequent to a downhill run. METHODS: Thirty-two recreationally active participants completed an incremental treadmill test in an HCS and a control running shoe (CON) for the determination of Oc and maximal performance. Subsequently, participants were pair matched and randomly assigned to one of the two footwear conditions to perform a moderate-intensity running bout before and 48 h after a 30-min downhill run designed to elicit muscle damage. RESULTS: Incremental treadmill test performance was improved (+5.7%; +1:16 min:ss; P < 0.01) in the HCS when assessed in the nondamaged state, relative to CON. This coincided with a significantly lower Oc (-3.2%; -6 mL·kg-1·km-1; P < 0.001) at a range of running speeds and an increase in the speed corresponding to 3 mM blood lactate (+3.2%; +0.4 km·h-1; P < 0.05). As anticipated, the downhill run resulted in significant changes in biochemical, histological, and perceptual markers of muscle damage, and a significant increase in Oc (+5.2%; 10.1 mL·kg-1·km-1) was observed 48 h post. In the presence of muscle damage, Oc was significantly lower in HCS (-4.6%; -10 mL·kg-1·km-1) compared with CON. CONCLUSIONS: These results indicate that HCS improved incremental exercise performance and Oc in the absence of muscle damage and show, for the first time, that despite worsening of Oc consequent to muscle damage, improved Oc in HCS is maintained.

Interaction of exercise bioenergetics with pacing behavior predicts track distance running performance.

The best possible finishing time for a runner competing in distance track events can be estimated from their critical speed (CS) and the finite amount of energy that can be expended above CS (D´). During tactical races with variable pacing, the runner with the "best" combination of CS and D´ and, therefore, the fastest estimated finishing time prior to the race, does not always win. We hypothesized that final race finishing positions depend on the relationships between the pacing strategies used, the athletes' initial CS, and their instantaneous D´ (i.e., D´ balance) as the race unfolds. Using publicly available data from the 2017 International Association of Athletics Federations (IAAF) World Championships men's 5,000-m and 10,000-m races, race speed, CS, and D´ balance were calculated. The correlation between D´ balance and actual finishing positions was nonsignificant using start-line values but improved to R2 > 0.90 as both races progressed. The D´ balance with 400 m remaining was strongly associated with both final 400-m split time and proximity to the winner. Athletes who exhausted their D´ were unable to hold pace with the leaders, whereas a high D´ remaining enabled a fast final 400 m and a high finishing position. The D´ balance model was able to accurately predict finishing positions in both a "slow" 5,000-m and a "fast" 10,000-m race. These results indicate that although CS and D´ can characterize an athlete's performance capabilities prior to the start, the pacing strategy that optimizes D´ utilization significantly impacts the final race outcome.NEW & NOTEWORTHY We show that the interaction between exercise bioenergetics and real-time pacing strategy predicts track distance running performance. Critical speed (CS) and the finite energy expended above CS (D´) can characterize an athlete's capabilities prior to the race start, but the pacing strategy that optimizes D´ utilization ultimately impacts whether a runner is in contention to win and whether a runner will have a fast final 400 m. Accordingly, D´ balance predicts final race finishing order.

The balance of muscle oxygen supply and demand reveals critical metabolic rate and predicts time to exhaustion.

We tested the hypothesis that during whole body exercise, the balance between muscle O2 supply and metabolic demand may elucidate intensity domains, reveal a critical metabolic rate, and predict time to exhaustion. Seventeen active, healthy volunteers (12 males, 5 females; 32 ± 2 yr) participated in two distinct protocols. Study 1 (n = 7) consisted of constant work rate cycling in the moderate, heavy, and severe exercise intensity domains with concurrent measures of pulmonary V̇o2 and local %SmO2 [via near-infrared spectroscopy (NIRS)] on quadriceps and forearm sites. Average %SmO2 at both sites displayed a domain-dependent response (P < 0.05). A negative %SmO2 slope was evident during severe-domain exercise but was positive during exercise below critical power (CP) at both muscle sites. In study 2 (n = 10), quadriceps and forearm site %SmO2 was measured during three continuous running trials to exhaustion and three intermittent intensity (ratio = 60 s severe: 30 s lower intensity) trials to exhaustion. Intensity-dependent negative %SmO2 slopes were observed for all trials (P < 0.05) and predicted zero slope at critical velocity. %SmO2 accurately predicted depletion and repletion of %D' balance on a second-by-second basis (R2 = 0.99, P < 0.05; both sites). Time to exhaustion predictions during continuous and intermittent exercise were either not different or better with %SmO2 [standard error of the estimate (SEE) < 20.52 s for quad, <44.03 s for forearm] versus running velocity (SEE < 65.76 s). Muscle O2 balance provides a dynamic physiological delineation between sustainable and unsustainable exercise (consistent with a "critical metabolic rate") and predicts real-time depletion and repletion of finite work capacity and time to exhaustion.NEW & NOTEWORTHY Dynamic muscle O2 saturation discriminates boundaries between exercise intensity domains, exposes a critical metabolic rate as the highest rate of steady state O2 supply and demand, describes time series depletion and repletion for work above critical power, and predicts time to exhaustion during severe domain whole body exercise. These results highlight the matching of O2 supply and demand as a primary determinant for sustainable exercise intensities from those that are unsustainable and lead to exhaustion.

Pannexin 1 channels control the hemodynamic response to hypoxia by regulating O2-sensitive extracellular ATP in blood.

Pannexin 1 (Panx1) channels export ATP and may contribute to increased concentration of the vasodilator ATP in plasma during hypoxia in vivo. We hypothesized that Panx1 channels and associated ATP export contribute to hypoxic vasodilation, a mechanism that facilitates the matching of oxygen delivery to metabolic demand of tissue. Male and female mice devoid of Panx1 (Panx1-/-) and wild-type controls (WT) were anesthetized, mechanically ventilated, and instrumented with a carotid artery catheter or femoral artery flow transducer for hemodynamic and plasma ATP monitoring during inhalation of 21% (normoxia) or 10% oxygen (hypoxia). ATP export from WT vs. Panx1-/-erythrocytes (RBC) was determined ex vivo via tonometer experimentation across progressive deoxygenation. Mean arterial pressure (MAP) was similar in Panx1-/- (n = 6) and WT (n = 6) mice in normoxia, but the decrease in MAP in hypoxia seen in WT was attenuated in Panx1-/- mice (-16 ± 9% vs. -2 ± 8%; P < 0.05). Hindlimb blood flow (HBF) was significantly lower in Panx1-/- (n = 6) vs. WT (n = 6) basally, and increased in WT but not Panx1-/- mice during hypoxia (8 ± 6% vs. -10 ± 13%; P < 0.05). Estimation of hindlimb vascular conductance using data from the MAP and HBF experiments showed an average response of 28% for WT vs. -9% for Panx1-/- mice. Mean venous plasma ATP during hypoxia was 57% lower in Panx1-/- (n = 6) vs. WT mice (n = 6; P < 0.05). Mean hypoxia-induced ATP export from RBCs from Panx1-/- mice (n = 8) was 82% lower than that from WT (n = 8; P < 0.05). Panx1 channels participate in hemodynamic responses consistent with hypoxic vasodilation by regulating hypoxia-sensitive extracellular ATP levels in blood.NEW & NOTEWORTHY Export of vasodilator ATP from red blood cells requires pannexin 1. Blood plasma ATP elevations in response to hypoxia in mice require pannexin 1. Hemodynamic responses to hypoxia are accompanied by increased plasma ATP in mice in vivo and require pannexin 1.

Physiological demands of running at 2-hour marathon race pace.

The requirements of running a 2-h marathon have been extensively debated but the actual physiological demands of running at ∼21.1 km/h have never been reported. We therefore conducted laboratory-based physiological evaluations and measured running economy (O2 cost) while running outdoors at ∼21.1 km/h, in world-class distance runners as part of Nike's "Breaking 2" marathon project. On separate days, 16 world-class male distance runners (age, 29 ± 4 yr; height, 1.72 ± 0.04 m; mass, 58.9 ± 3.3 kg) completed an incremental treadmill test for the assessment of V̇O2peak, O2 cost of submaximal running, lactate threshold and lactate turn-point, and a track test during which they ran continuously at 21.1 km/h. The laboratory-determined V̇O2peak was 71.0 ± 5.7 mL/kg/min with lactate threshold and lactate turn-point occurring at 18.9 ± 0.4 and 20.2 ± 0.6 km/h, corresponding to 83 ± 5% and 92 ± 3% V̇O2peak, respectively. Seven athletes were able to attain a steady-state V̇O2 when running outdoors at 21.1 km/h. The mean O2 cost for these athletes was 191 ± 19 mL/kg/km such that running at 21.1 km/h required an absolute V̇O2 of ∼4.0 L/min and represented 94 ± 3% V̇O2peak. We report novel data on the O2 cost of running outdoors at 21.1 km/h, which enables better modeling of possible marathon performances by elite athletes. Using the value for O2 cost measured in this study, a sub 2-h marathon would require a 59 kg runner to sustain a V̇O2 of approximately 4.0 L/min or 67 mL/kg/min.NEW & NOTEWORTHY We report the physiological characteristics and O2 cost of running overground at ∼21.1 km/h in a cohort of the world's best male distance runners. We provide new information on the absolute and relative O2 uptake required to run at 2-h marathon pace.

Dynamics of the power-duration relationship during prolonged endurance exercise and influence of carbohydrate ingestion.

We tested the hypotheses that the parameters of the power-duration relationship, estimated as the end-test power (EP) and work done above EP (WEP) during a 3-min all-out exercise test (3MT), would be reduced progressively after 40 min, 80 min, and 2 h of heavy-intensity cycling and that carbohydrate (CHO) ingestion would attenuate the reduction in EP and WEP. Sixteen participants completed a 3MT without prior exercise (control), immediately after 40 min, 80 min, and 2 h of heavy-intensity exercise while consuming a placebo beverage, and also after 2 h of heavy-intensity exercise while consuming a CHO supplement (60 g/h CHO). There was no difference in EP measured without prior exercise (260 ± 37 W) compared with EP after 40 min (268 ± 39 W) or 80 min (260 ± 40 W) of heavy-intensity exercise; however, after 2 h EP was 9% lower compared with control (236 ± 47 W; P < 0.05). There was no difference in WEP measured without prior exercise (17.9 ± 3.3 kJ) compared with after 40 min of heavy-intensity exercise (16.1 ± 3.3 kJ), but WEP was lower (P < 0.05) than control after 80 min (14.7 ± 2.9 kJ) and 2 h (13.8 ± 2.7 kJ). Compared with placebo, CHO ingestion negated the reduction of EP following 2 h of heavy-intensity exercise (254 ± 49 W) but had no effect on WEP (13.5 ± 3.4 kJ). These results reveal a different time course for the deterioration of EP and WEP during prolonged endurance exercise and indicate that EP is sensitive to CHO availability.NEW & NOTEWORTHY The parameters of the power-duration relationship [critical power (CP) and the curvature constant (W')] have typically been considered to be static. Here we report the time course for reductions in CP and W', as estimated with the 3-min all-out cycle test, during 2 h of heavy-intensity exercise. We also show that carbohydrate ingestion during exercise preserves CP, but not W', without altering muscle glycogen depletion. These results provide new mechanistic and practical insight into the power-duration curve and its relationship to exercise-related fatigue development.

Changes in the power-duration relationship following prolonged exercise: estimation using conventional and all-out protocols and relationship with muscle glycogen.

It is not clear how the parameters of the power-duration relationship [critical power (CP) and W'] are influenced by the performance of prolonged endurance exercise. We used severe-intensity prediction trials (conventional protocol) and the 3-min all-out test (3MT) to measure CP and W' following 2 h of heavy-intensity cycling exercise and took muscle biopsies to investigate possible relationships to changes in muscle glycogen concentration ([glycogen]). Fourteen participants completed a rested 3MT to establish end-test power (Control-EP) and work done above EP (Control-WEP). Subsequently, on separate days, immediately following 2 h of heavy-intensity exercise, participants completed a 3MT to establish Fatigued-EP and Fatigued-WEP and three severe-intensity prediction trials to the limit of tolerance (Tlim) to establish Fatigued-CP and Fatigued-W'. A muscle biopsy was collected immediately before and after one of the 2-h exercise bouts. Fatigued-CP (256 ± 41 W) and Fatigued-EP (256 ± 52 W), and Fatigued-W' (15.3 ± 5.0 kJ) and Fatigued-WEP (14.6 ± 5.3 kJ), were not different (P > 0.05) but were ~11% and ~20% lower than Control-EP (287 ± 46 W) and Control-WEP (18.7 ± 4.7 kJ), respectively (P < 0.05). The change in muscle [glycogen] was not significantly correlated with the changes in either EP (r = 0.19) or WEP (r = 0.07). The power-duration relationship is adversely impacted by prolonged endurance exercise. The 3MT provides valid estimates of CP and W' following 2 h of heavy-intensity exercise, but the changes in these parameters are not primarily determined by changes in muscle [glycogen].

Critical Velocity during Intermittent Running with Changes of Direction.

PURPOSE: We tested the hypothesis that critical velocity (CV) during intermittent running with changes of direction is reliably and accurately identified from a simple shuttle field test. We also tested the hypothesis that CV during intermittent running with changes of direction running is not equivalent to continuous linear running. METHODS: Young adults performed a custom shuttle test of intermittent sprint running to reveal CV. Sprints were 18.3 m per direction, with rest between sprints of 15 s for 3 min, 10 s for 2 min, and no rest for 2 min (7 min total). To test reliability, the CV shuttle test (CVST) was performed twice. To test validity, blood lactate was assessed during two separate trials inclusive of 5% above or below CVST end velocity. To explore task specificity, CV during CVST was compared to CV obtained from three linear running time trials. RESULTS: Total distance and CSVT end test velocity were similar between visits (864 ± 21 m and 3.23 ± 0.13 m·s vs 900 ± 30 m and 3.21 ± 0.15 m·s, respectively). At 5% above CVST end velocity, all subjects failed to complete 20 min and had unstable blood lactate values. A steady state blood lactate profile was observed during trials 5% below end velocity and all subjects completed the trial. The CV from the CVST was lower than the CV from linear running (△ -17% ± 6%), highlighting the importance of test specificity for threshold determination. CONCLUSIONS: The CVST provides a reliable and accurate determination of CV and can be used by coaches, athletes, and trainers to better understand the physiological impact specific to practice or competitions involving intermittent change of direction running.

Effects of Two Hours of Heavy-Intensity Exercise on the Power-Duration Relationship.

INTRODUCTION: Changes in the parameters of the power-time relationship (critical power (CP) and W') during endurance exercise would have important implications for performance. We tested the hypotheses that CP and W', estimated using the end-test power (EP) and the work done above EP (WEP), respectively, during a the 3-min all-out test (3MT), can be reliably determined, and would be lower, after completing 2 h of heavy-intensity exercise. METHODS: In study 1, six cyclists completed a 3MT immediately after 2 h of heavy-intensity exercise on two occasions to establish the reliability of EP and WEP. In study 2, nine cyclists completed a control 3MT, and a fatigued 3MT and constant power output tests to 30 min or the limit of tolerance (Tlim) below and above F-EP after 2 h of heavy-intensity exercise. RESULTS: In study 1, EP (273 ± 52 vs 276 ± 58 W) and WEP (12.4 ± 4.3 vs 12.8 ± 4.3 kJ) after 2 h of heavy-intensity exercise were not different (P > 0.05) and were highly correlated (r = 0.99; P < 0.001). In study 2, both EP (F-EP: 282 ± 52 vs C-EP: 306 ± 56 W; P < 0.01) and WEP (F-WEP: 14.7 ± 4.9 vs C-WEP: 18.3 ± 4.1 kJ; P < 0.05) were lower after 2-h heavy-intensity exercise. However, maximum O2 uptake was not achieved during exercise >F-EP and Tlim was shorter than 30 min during exercise

Endothelium-dependent vasodilatory signalling modulates α1 -adrenergic vasoconstriction in contracting skeletal muscle of humans.

KEY POINTS: 'Functional sympatholysis' describes the ability of contracting skeletal muscle to attenuate sympathetic vasoconstriction, and is critical to ensure proper blood flow and oxygen delivery to metabolically active skeletal muscle. The signalling mechanism responsible for sympatholysis in healthy humans is unknown. Evidence from animal models has identified endothelium-derived hyperpolarization (EDH) as a potential mechanism capable of attenuating sympathetic vasoconstriction. In this study, increasing endothelium-dependent signalling during exercise significantly enhanced the ability of contracting skeletal muscle to attenuate sympathetic vasoconstriction in humans. This is the first study in humans to identify endothelium-dependent regulation of sympathetic vasoconstriction in contracting skeletal muscle, and specifically supports a role for EDH-like vasodilatory signalling. Impaired functional sympatholysis is a common feature of cardiovascular ageing, hypertension and heart failure, and thus identifying fundamental mechanisms responsible for sympatholysis is clinically relevant. ABSTRACT: Stimulation of α-adrenoceptors elicits vasoconstriction in resting skeletal muscle that is blunted during exercise in an intensity-dependent manner. In humans, the underlying mechanisms remain unclear. We tested the hypothesis that stimulating endothelium-dependent vasodilatory signalling will enhance the ability of contracting skeletal muscle to blunt α1 -adrenergic vasoconstriction. Changes in forearm vascular conductance (FVC; Doppler ultrasound, brachial intra-arterial pressure via catheter) to local intra-arterial infusion of phenylephrine (PE; α1 -adrenoceptor agonist) were calculated during (1) infusion of the endothelium-dependent vasodilators acetylcholine (ACh) and adenosine triphosphate (ATP), the endothelium-independent vasodilator (sodium nitroprusside, SNP), or potassium chloride (KCl) at rest; (2) mild or moderate intensity handgrip exercise; and (3) combined mild exercise + ACh, ATP, SNP, or KCl infusions in healthy adults. Robust vasoconstriction to PE was observed during vasodilator infusion alone and mild exercise, and this was blunted during moderate intensity exercise (ΔFVC: -34 ± 4 and -34 ± 3 vs. -13 ± 2%, respectively, P < 0.05). Infusion of ACh or ATP during mild exercise significantly attenuated PE vasoconstriction similar to levels observed during moderate exercise (ACh: -3 ± 4; ATP: -18 ± 4%). In contrast, infusion of SNP or KCl during mild exercise did not attenuate PE-mediated vasoconstriction (-32 ± 5 and -46 ± 3%). To further study the role of endothelium-dependent hyperpolarization (EDH), ACh trials were repeated with combined nitric oxide synthase and cyclooxygenase inhibition. Here, PE-mediated vasoconstriction was blunted at rest (blockade: -20 ± 5 vs. CONTROL: -31 ± 3% vs.; P < 0.05) and remained blunted during exercise (blockade: -15 ± 5 vs. CONTROL: -14 ± 5%). We conclude that stimulation of EDH-like vasodilatation can blunt α1 -adrenergic vasoconstriction in contracting skeletal muscle of humans.

Contracting human skeletal muscle maintains the ability to blunt α1 -adrenergic vasoconstriction during KIR channel and Na(+) /K(+) -ATPase inhibition.

KEY POINTS: During exercise there is a balance between vasoactive factors that facilitate increases in blood flow and oxygen delivery to the active tissue and the sympathetic nervous system, which acts to limit muscle blood flow for the purpose of blood pressure regulation. Functional sympatholysis describes the ability of contracting skeletal muscle to blunt the stimulus for vasoconstriction, yet the underlying signalling of this response in humans is not well understood. We tested the hypothesis that activation of inwardly rectifying potassium channels and the sodium-potassium ATPase pump, two potential vasodilator pathways within blood vessels, contributes to the ability to blunt α1 -adrenergic vasoconstriction. Our results show preserved blunting of α1 -adrenergic vasconstriction despite blockade of these vasoactive factors. Understanding this complex phenomenon is important as it is impaired in a variety of clinical populations. ABSTRACT: Sympathetic vasoconstriction in contracting skeletal muscle is blunted relative to that which occurs in resting tissue; however, the mechanisms underlying this 'functional sympatholysis' remain unclear in humans. We tested the hypothesis that α1 -adrenergic vasoconstriction is augmented during exercise following inhibition of inwardly rectifying potassium (KIR ) channels and Na(+) /K(+) -ATPase (BaCl2  + ouabain). In young healthy humans, we measured forearm blood flow (Doppler ultrasound) and calculated forearm vascular conductance (FVC) at rest, during steady-state stimulus conditions (pre-phenylephrine), and after 2 min of phenylephrine (PE; an α1 -adrenoceptor agonist) infusion via brachial artery catheter in response to two different stimuli: moderate (15% maximal voluntary contraction) rhythmic handgrip exercise or adenosine infusion. In Protocol 1 (n = 11 subjects) a total of six trials were performed in three conditions: control (saline), combined enzymatic inhibition of nitric oxide (NO) and prostaglandin (PG) synthesis (l-NMMA + ketorolac) and combined inhibition of NO, PGs, KIR channels and Na(+) /K(+) -ATPase (l-NMMA + ketorolac + BaCl2  + ouabain). In Protocol 2 (n = 6) a total of four trials were performed in two conditions: control (saline), and combined KIR channel and Na(+) /K(+) -ATPase inhibition. All trials occurred after local ß-adrenoceptor blockade (propranolol). PE-mediated vasoconstriction was calculated (%ΔFVC) in each condition. Contrary to our hypothesis, despite attenuated exercise hyperaemia of ∼30%, inhibition of KIR channels and Na(+) /K(+) -ATPase, combined with inhibition of NO and PGs (Protocol 1) or alone (Protocol 2) did not enhance α1 -mediated vasoconstriction during exercise (Protocol 1: -27 ± 3%; P = 0.2 vs. control, P = 0.4 vs. l-NMMA + ketorolac; Protocol 2: -21 ± 7%; P = 0.9 vs. control). Thus, contracting human skeletal muscle maintains the ability to blunt α1 -adrenergic vasoconstriction during combined KIR channel and Na(+) /K(+) -ATPase inhibition.

Intravascular ATP and the regulation of blood flow and oxygen delivery in humans.

Regulation of vascular tone is a complex response that integrates multiple signals that allow for blood flow and oxygen supply to match oxygen demand appropriately. Here, we discuss the potential role of intravascular adenosine triphosphate (ATP) as a primary factor in these responses and put forth the hypothesis that deficient ATP release contributes to impairments in vascular control exhibited in aged and diseased populations.

Restoration of intracellular ATP production in banked red blood cells improves inducible ATP export and suppresses RBC-endothelial adhesion.

Transfusion of banked red blood cells (RBCs) has been associated with poor cardiovascular outcomes. Storage-induced alterations in RBC glycolytic flux, attenuated ATP export, and microvascular adhesion of transfused RBCs in vivo could contribute, but the underlying mechanisms have not been tested. We tested the novel hypothesis that improving deoxygenation-induced metabolic flux and the associated intracellular ATP generation in stored RBCs (sRBCs) results in an increased extracellular ATP export and suppresses microvascular adhesion of RBCs to endothelium in vivo following transfusion. We show deficient intracellular ATP production and ATP export by human sRBCs during deoxygenation (impairments ~42% and 49%, respectively). sRBC pretreatment with a solution containing glycolytic intermediate/purine/phosphate precursors (i.e., "PIPA") restored deoxygenation-induced intracellular ATP production and promoted extracellular ATP export (improvement ~120% and 50%, respectively). In a nude mouse model of transfusion, adhesion of human RBCs to the microvasculature in vivo was examined. Only 2% of fresh RBCs (fRBCs) transfused adhered to the vascular wall, compared with 16% of sRBCs transfused. PIPA pretreatment of sRBCs significantly reduced adhesion to just 5%. In hypoxia, adhesion of sRBCs transfused was significantly augmented (up to 21%), but not following transfusion of fRBCs or PIPA-treated sRBCs (3.5% or 6%). Enhancing the capacity for deoxygenation-induced glycolytic flux within sRBCs increases their ability to generate intracellular ATP, improves the inducible export of extracellular anti-adhesive ATP, and consequently suppresses adhesion of stored, transfused RBCs to the vascular wall in vivo.

Randomized study of washing 40- to 42-day-stored red blood cells.

BACKGROUND: Pretransfusion washing of red blood cells (RBCs) stored for a longer duration may have theoretical advantages but few data exist to support this practice. In many hospital settings, use of a point-of-care cell washer could conceivably be used to quickly wash allogeneic RBCs before transfusion. The purpose of this preliminary study was to compare a point-of-care device with a common blood bank device for washing longer-stored RBCs. STUDY DESIGN AND METHODS: Forty RBC units stored for 40 to 42 days were randomized to washing with the COBE 2991 device (Terumo BCT; FDA-cleared for washing stored RBCs) or the Cell Saver Elite (Haemonetics; FDA-cleared point-of-care device for processing and washing fresh autologous shed whole blood). Supernatant and unit RBCs from unwashed (baseline) and washed blood were assayed for potassium, lactate, intracellular ATP, percentage of RBC recovery, cell-free hemoglobin, RBC microparticles, and RBCs were examined for susceptibility to hemolysis by physical stress. RESULTS: Both devices recovered a high percentage of RBCs and efficiently removed extracelluar potassium. Washing with the Elite resulted in significant increases in cell-free Hb, percent hemolysis, and RBC microparticle production, whereas washing with the COBE 2991 did not (fold Δ = 2.1 vs. 1.0, 4.6 vs. 1.2, 2.0 vs. 1.1, respectively; p < 0.05). Hemolysis induced by physical stress was not altered by washing. CONCLUSION: Although point-of-care washing of longer-stored RBCs is appealing, these preliminary data suggest that transfusion of washed, longer-stored units could result in potentially greater exposure to plasma free Hb. More data are needed before this practice can be routinely recommended.