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Dietary nitrate (NO 3 -) supplementation via beetroot juice (BR) has been reported to lower oxygen cost (i.e., increased exercise efficiency) and speed up oxygen uptake (VO 2) kinetics in untrained and moderately trained individuals, particularly during conditions of low oxygen availability (i.e., hypoxia). However, the effects of multiple-day, high dose (12.4 mmol NO 3- per day) BR supplementation on exercise efficiency and VO 2 kinetics during normoxia and hypoxia in well-trained individuals are not resolved. In a double-blinded, randomized crossover study, 12 well-trained cyclists (66.4 ± 5.3 ml min -1∙kg -1) completed three transitions from rest to moderate-intensity (~70% of gas exchange threshold) cycling in hypoxia and normoxia with supplementation of BR or nitrate-depleted BR as placebo. Continuous measures of VO 2 and muscle (vastus lateralis) deoxygenation (ΔHHb, using near-infrared spectroscopy) were acquired during all transitions. Kinetics of VO 2 and deoxygenation (ΔHHb) were modeled using mono-exponential functions. Our results showed that BR supplementation did not alter the primary time constant for VO 2 or ΔHHb during the transition from rest to moderate-intensity cycling. While BR supplementation lowered the amplitude of the VO 2 response (2.1%, p = 0.038), BR did not alter steady state VO 2 derived from the fit (p = 0.258), raw VO 2 data (p = 0.231), moderate intensity exercise efficiency (p = 0.333) nor steady state ΔHHb (p = 0.224). Altogether, these results demonstrate that multiple-day, high-dose BR supplementation does not alter exercise efficiency or oxygen uptake kinetics during normoxia and hypoxia in well-trained athletes. more...

Dietary nitrate (NO3-) supplementation via beetroot juice (BR) is known to improve endurance performance in untrained and moderately trained individuals. However, conflicting results exist in well-trained individuals. Evidence suggests that the effects of NO3- are augmented during conditions of reduced oxygen availability (e.g., hypoxia), thereby increasing the probability of performance improvements for well-trained athletes in hypoxia vs. normoxia. This randomized, double-blinded, counterbalanced-crossover study examined the effects of 7 days of BR supplementation with 12.4 mmol NO3- per day on 10-km cycling time trial (TT) performance in 12 well-trained cyclists in normoxia (N) and normobaric hypoxia (H). Linear mixed models for repeated measures revealed increases in plasma NO3- and NO2- after supplementation with BR (both p more...

Noninvasive imaging of oxygen uptake may provide a useful tool for the quantification of energy expenditure during human locomotion. A novel thermal imaging method (optical flow) was validated against indirect calorimetry for the estimation of energy expenditure during human walking and running.Fourteen endurance-trained subjects completed a discontinuous incremental exercise test on a treadmill. Subjects performed 4-min intervals at 3, 5, and 7 km·h (walking) and at 8, 10, 12, 14, 16, and 18 km·h (running) with 30 s of rest between intervals. Heart rate, gas exchange, and mean accelerations of ankle, thigh, wrist, and hip were measured throughout the exercise test. A thermal camera (30 frames per second) was used to quantify optical flow, calculated as the movements of the limbs relative to the trunk (internal mechanical work) and vertical movement of the trunk (external vertical mechanical work).Heart rate, gross oxygen uptake (mL·kg·min) together with gross and net energy expenditure (J·kg·min) rose with increasing treadmill velocities, as did optical flow measurements and mean accelerations (g) of ankle, thigh, wrist, and hip. Oxygen uptake was linearly correlated with optical flow across all exercise intensities (R = 0.96, P0.0001; V˙O2 [mL·kg·min] = 7.35 + 9.85 × optical flow [arbitrary units]). Only 3-4 s of camera recording was required to estimate an optical flow value at each velocity.Optical flow measurements provide an accurate estimation of energy expenditure during horizontal walking and running. The technique offers a novel experimental method of estimating energy expenditure during human locomotion, without use of interfering equipment attached to the subject. more...

Aim: Mitochondrial dysfunction has been suggested to play a significant role in obesity and insulin resistance. The aim of the present study was to investigate if changes in obesity and insulin resistance were related to similar changes in mitochondrial capacity for hydrogenperoxide release in Zucker diabetic fatty rats and their lean littermates. Methods: Thirty-four rats were used in this study. Rats were either lean or obese Zucker rats killed at 5–6 (young) or 12–14 (adults) weeks of age. Mitochondria were isolated from soleus muscles; respiration and release of hydrogenperoxide were determined and related to citrate synthase activity to determine intrinsic mitochondrial function. Mitochondrial-specific super-oxide dismuthase (MnSOD) protein content was determined in isolated mitochondria and muscle homogenate. Catalase protein content was determined in muscle homogenate. Results: Young lean and obese rats had a higher mitochondrial respiration when using palmitoyl-l-carnitine as substrate compared with adult lean and obese rats. The obese strain had higher mitochondrial hydrogenperoxide release but only in the adult animals. In both lean and obese animals, increased age was associated with increased mitochondrial hydrogenperoxide release. MnSOD tended to be higher in the obese strain in the isolated mitochondria. Regardless of age, catalase protein content was significantly lower in the obese rats. Conclusions: This study shows that the augmented increase in obesity and insulin resistance seen in Zucker diabetic fatty rats is associated with increased capacity for mitochondrial hydrogenperoxide release. more...

We investigated the effects of insulin and adrenaline on the rate of glycogen synthesis in skeletal muscles after electrical stimulation in vitro. The contractile activity decreased the glycogen concentration by 62%. After contractile activity, the glycogen stores were fully replenished at a constant and high rate for 3 h when 10 m-i.u./ml insulin was present. In the absence of insulin, only 65% of the initial glycogen stores was replenished. Adrenaline decreased insulin-stimulated glycogen synthesis. Surprisingly, adrenaline did not inhibit glycogen synthesis stimulated by glycogen-depleting contractile activity. In agreement with this, the fractional activity of glycogen synthase was high when adrenaline was present after exercise, whereas adrenaline decreased the fractional activity of glycogen synthase to a low level during stimulation with insulin. Furthermore, adrenaline activated glycogen phosphorylase almost completely during stimulation with insulin, whereas a much lower activation of glycogen phosphorylase was observed after contractile activity. Thus adrenaline does not inhibit contraction-stimulated glycogen synthesis. more...

Purpose: To compare the effects of three types of intensive run training on running economy (RE) during exhaustive running and to establish possible relationships with changes in ventilatory function and/or muscle fiber type distribution. Methods: Thirty-six male recreational runners were divided into three groups and assigned to either exhaustive distance training (DT), long- interval training (LIT), or short-interval training (SIT) three times 20-30 min · wk-1 for 6 wk. V̇O(2max) and RE were measured during treadmill running before and after training. Muscle fiber type distribution of the vastus lateralis muscle was established from biopsy material. Results: V̇O(2max) (L · min-1) increased by 5.9% (P < 0.0001), 6.0% (P < 0.0001), and 3.6% (P < 0.01) in DT, LIT, and SIT, respectively, and running speed at V̇O(2max) by 9% (P < 0.0001), 10% (P < 0.0001), and 4% (P < 0.05), respectively. Time-to-exhaustion at 87% of pretraining V̇O(2max) (mean 3.83 m · s-1) increased by 94% in DT (P < 0.0001), 67% in LIT (P < 0.0001), and 65% in SIT (P < 0.001). Running economy improved by 3.1% in DT (P < 0.05), 3.0% in LIT (P < 0.01), and 0.9% in SIT (NS); pulmonary ventilation (V̇E) was on average 11 L · min-1 lower following training (P < 0.0001). The individual decrements in V̇E correlated with improvements in RE (r = 0.77; P < 0.0001) and may account for 25-70% of the decrease in aerobic demand. Muscle fiber composition, and respiratory exchange ratio, stride length, and stride frequency during running were unaltered with training. Conclusion: Recreational runners can improve RE and aerobic run performance by exchanging parts of their conventional aerobic distance training with intensive distance or long-interval running, whereas short-interval running is less efficient. The improvement in RE may relate to reduced ventilatory demands. Muscle fiber type distribution was unaltered with training and showed no association with RE. more...

For most people, carbohydrate in polymer form is the main energy source in the diet. For uptake and transport between cells, carbohydrates are broken down to monosaccharides and glucose is the main sugar in the blood. A number of different glucose transporters (GLUTs) transport glucose across cell membranes. Glucose is stored as glycogen particles in skeletal muscles (∼500g) and the liver (∼100g). Glucose is utilised continuously, but the glucose concentration in blood remains relatively stable although food is consumed in few larger meals with periods without food intake; the liver release glucose into the blood in periods without intestinal glucose absorption. At rest, glucose utilisation is low (0.1g·min−1), but can increase above 3g·min−1 of glucose during exercise in well-trained subjects; glycogen in the active muscles is the main substrate and glycogen depletion causes fatigue. The glycogen stores are limited and excess glucose is converted into fat, and de novo lipid synthesis from superfluous glucose is a major reason for obesity and type 2 diabetes. Exercise decreases glycogen content in skeletal muscles; exercise also increases insulin sensitivity and prevents development of type 2 diabetes. more...

Fatty acid metabolism is influenced by training and diet with exercise training mediating this through activation of nuclear hormone receptor peroxisome proliferator-activated receptor alpha (PPARalpha) in skeletal muscle. This study investigated the effect of training and high fat or normal diet on PPARalpha expression in human skeletal muscle. Thirteen men trained one leg (T) four weeks (31.5 h in total), while the other leg (UT) served as control. During the 4 weeks six subjects consumed high fat (FAT) diet and seven subjects maintained a normal (CHO) diet. Biopsies were obtained from vastus lateralis muscle in both legs before and after training. After the biopsy, one-leg extension exercise was performed in random order with both legs 30 min at 95% of workload max. A training effect was evident as citrate synthase activity increased (P0.05) by 15% in the trained, but not the control leg in both groups. During exercise respiratory exchange ratio was lower in FAT (0.86 +/- 0.01, 0.83 +/- 0.01, mean +/- SEM) than CHO (0.96 +/- 0.02, 0.94 +/- 0.03) and in UT than T legs, respectively. The PPARalpha protein (144 +/- 44, 104 +/- 28, 79 +/- 15, 79 +/- 14, % of pre level) and PPARalpha mRNA (69 +/- [2, 2], 78 +/- [7, 6], 92 +/- [22, 18], 106 +/- [21, 18], % of pre level, geometric mean +/- SEM) expression remained unchanged by diet and training in FAT (UT, T) and CHO (UT, T), respectively. After the training and diet CS, HAD, PPARalpha, UCP2, UCP3 and mFABP mRNA content remained unchanged, whereas GLUT4 mRNA was lower in both groups and LDHA mRNA was lower (P0.05) only in FAT.4 weeks one leg knee extensor training did not affect PPARalpha protein or mRNA expression. Furthermore, higher fat oxidation during exercise after fat rich diet was not accompanied by an increased PPARalpha protein or mRNA expression after 4 weeks. more...

Accumulation of acyl-CoA is hypothesized to be involved in development of insulin resistance. Acyl-CoA binds to acyl-CoA binding protein (ACBP) with high affinity, and therefore knowledge about ACBP concentration is important for interpreting acyl-CoA data. In the present study, we used a sandwich enzyme-linked immunosorbent assay to quantify ACBP concentration in different muscle fiber types. Furthermore, ACBP concentration was compared in muscles from lean and obese Zucker rats. Expression of ACBP was highest in the slow-twitch oxidative soleus muscle and lowest in the fast-twitch glycolytic white gastrocnemius (0.46 ± 0.02 and 0.16 ± 0.005 μg/mg protein, respectively). Expression of ACBP was soleus > red gastrocnemius > extensor digitorum longus > white gastrocnemius. Similar fiber type differences were found for carnitine palmitoyl transferase (CPT)-1, and a correlation was observed between ACBP and CPT-1. Muscles from obese Zucker rats had twice the triglyceride content, had approximately twice the long-chain acyl CoA content, and were severely insulin resistant. ACBP concentration was ∼30% higher in all muscles from obese rats. Activities of CPT-1 and 3-hydroxy-acyl-CoA dehydrogenase were increased in muscles from obese rats, whereas citrate synthase activity was similar. In conclusion, ACBP expression is fiber type-specific with the highest concentration in oxidative muscles and the lowest in glycolytic muscles. The 90% increase in the concentration of acyl-CoA in obese Zucker muscle compared with only a 30% increase in the concentration of ACBP supports the hypothesis that an increased concentration of free acyl-CoA is involved in the development of insulin resistance. more...

The metabolic active form of free fatty acids, long-chain acyl-coenzyme A (lc-acyl-CoA), binds to its 10-kDa binding protein with high affinity. In the present study, we investigated the content of lc-acyl-CoA binding protein (ACBP) in different skeletal muscle fibre types. Soleus had the highest expression of ACBP (0.33±0.02 μg mg protein-1) and the content was as high as in heart muscle. The content in mixed gastrocnemius (0.27±0.02 gg mg protein-1), extensor digitorum longus (0.21±0.01 gg mg protein-1) and white gastrocnemius (0.16±0.01 μg mg protein-1) were lower than in soleus and differed from each other (P more...

In the present study, the relationship between the pattern of electrical stimulation and glucose uptake was investigated in slow-twitch muscles (soleus) and fast-twitch muscles (epitrochlearis) from Wistar rats. Muscles were stimulated electrically for 30 min in vitro with either single pulses (frequencies varied between 0.8 and 15 Hz) or with 200-ms trains (0.1–2 Hz). Glucose uptake (measured with tracer amount of 2-[3H]deoxyglucose) increased with increasing number of impulses whether delivered as single pulses or as short trains. The highest glucose uptake achieved with short tetanic contractions was similar in soleus and epitrochlearis (10.9 ± 0.7 and 12.0 ± 0.8 mmol · kg dry wt−1· 30 min−1, respectively). Single pulses, on the other hand, increased contraction-stimulated glucose uptake less in soleus than in epitrochlearis (7.5 ± 1.1 and 11.7 ± 0.5 mmol · kg dry wt−1· 30 min−1, respectively; P < 0.02). Glucose uptake correlated with glycogen breakdown in soleus ( r = 0.84, P < 0.0001) and (epitrochlearis: r = 0.91, P < 0.0001). Contraction-stimulated glucose uptake also correlated with breakdown of ATP and PCr and with reduction in force. Our data suggest that metabolic stress mediates contraction-stimulated glucose uptake. more...

Udgivelsesdato: 2000-Nov Prompted by our recent observations that GLUT-1 is expressed in fetal muscles, but not in adult muscle fibers, we decided to investigate whether GLUT-1 expression could be reactivated. We studied different stimuli concerning their ability to induce GLUT-1 expression in mature human skeletal muscle fibers. Metabolic stress (obesity, non-insulin-dependent diabetes mellitus), contractile activity (training), and conditions of de- and reinnervation (amyotrophic lateral sclerosis) could not induce GLUT-1 expression in human muscle fibers. However, regenerating muscle fibers in polymyositis expressed GLUT-1. In contrast to GLUT-1, GLUT-4 was expressed in all investigated muscle fibers. Although the significance of GLUT-1 in adult human muscle fibers appears limited, GLUT-1 may be of importance for the glucose supplies in immature and regenerating muscle. more...

Thirty‐nine moderately endurance trained males increased their normal training programme of 2.2 h week‐1 with an average training intensity of 65 % of maximum heart rate (HRmax) to 2.7 h week‐1 and a mean intensity of 78% of HRmax. Performance tests and measurements of the total concentrations of Na,K‐ATPase (3H‐ouabain binding) and Ca‐ATPase, fibre type distribution and fibre area were performed in biopsies from the vastus lateralis muscle before and after increased training. The 6 weeks of training elevated Vo2max from 54.9 + 3.1 to 58.3±3.0 ml 02 min‐1 kg‐1 (P < 0.0001). Exercise time to exhaustion at 86% of Fo2max (pre‐training) increased from 35 ±8 to 61 + 17 min (P < 0.0001). The concentration of Ca‐ATPase was unaffected by the intensified training (6.74 ± 1.03 vs. 6.68+ 1.07 nmol g wet wt‐1), but the concentration of Na,K‐ATPase increased from 307±43 to 354 + 59 pmol g wet wt ' (P < 0.0001). The relative distribution of FT‐fibres was correlated with the concentration of Ca‐ATPase (r = 0.72, P < 0.0001). The data support the view that intensive training induces an upregulation of the concentration of skeletal muscle Na,K‐ATPase, but no change in the total capacity for reaccumulation of Ca2+ into the SR. There was no correlation between the concentrations of Na,K‐ATPase, Ca‐ATPase and indices of endurance performance. more...