Jostein Hallén

Evidence against a Beneficial Effect of Irisin in Humans

Brown adipose tissue has gained interest as a potential target to treat obesity and metabolic diseases. Irisin is a newly identified hormone secreted from skeletal muscle enhancing browning of white fat cells, which improves systemic metabolism by increasing energy expenditure in mice. The discovery of irisin raised expectations of its therapeutic potential to treat metabolic diseases. However, the effect of irisin in humans is unclear. Analyses of genomic DNA, mRNA and expressed sequence tags revealed that FNDC5, the gene encoding the precursor of irisin, is present in rodents and most primates, but shows in humans a mutation in the conserved start codon ATG to ATA. HEK293 cells transfected with a human FNDC5 construct with ATA as start codon resulted in only 1% full-length protein compared to human FNDC5 with ATG. Additionally, in vitro contraction of primary human myotubes by electrical pulse stimulation induced a significant increase in PGC1α mRNA expression. However, FNDC5 mRNA level was not altered. FNDC5 mRNA expression in muscle biopsies from two different human exercise studies was not changed by endurance or strength training. Preadipocytes isolated from human subcutaneous adipose tissue exhibited differentiation to brite human adipocytes when incubated with bone morphogenetic protein (BMP) 7, but neither recombinant FNDC5 nor irisin were effective. In conclusion, our findings suggest that it is rather unlikely that the beneficial effect of irisin observed in mice can be translated to humans.

Hormonal responses to high- and moderate-intensity strength exercise

Abstract The hormonal responses of nine male, strength athletes to strength exercise were examined. The athletes performed one moderate- and one high-intensity strength exercise workout. In the high-intensity workout, the load was 100% of each subjects three-repetition maximum (3-RM) for squats and front squats, and 100% of each subjects six-repetition maximum (6-RM) for leg extensions. In the moderate-intensity workout, the load was 70% of the high-intensity protocol. Rest periods between sets were 4–6 min for both workouts. Blood samples were taken before, 30 min into, and every 15 min for the 1st h after exercise, and then 3, 7, 11, 22 and 33 h after exercise, thus allowing examination of both the acute and prolonged hormonal responses. Blood samples were analyzed for testosterone, luteinizing hormone (LH), follicle stimulating hormone (FSH), cortisol, adrenocorticotrophic hormone (ACTH), growth hormone (GH), insulin-like growth factor (IGF-1), insulin, sex hormone binding globulin, creatine kinase, total protein, glucose and lactate. The acute responses of testosterone and cortisol were greater during the high-intensity protocol as compared to the moderate-intensity protocol. The cortisol response was associated with an increase in ACTH concentration. LH and FSH showed no response to either protocol. Acute GH responses were not different between protocols. There were great inter-individual differences in acute GH responses to both protocols. There were no significant differences between protocols with regard to prolonged responses for any hormone. In both trials, IGF-1 concentrations were significantly lower at 0800 hours the morning after exercise as compared to concentrations found at 0800 hours the morning before exercise. The mechanisms responsible for reducing IGF-1 concentration in these trials are unclear, and it is not known if this reduction observed 22 hours after exercise is of physiological significance.

Effect of rhepo administration on serum levels of stfr and cycling performance

PURPOSE We assessed the possibility of using soluble transferrin receptor (sTfR) as an indicator of doping with recombinant erythropoietin (rhEPO). METHODS A double-blind, placebo-controlled study was conducted with the administration of 5,000 U of rhEPO (N = 10) or placebo (N = 10) three times weekly (181-232 U x kg(-1) x wk-1) for 4 wk to male athletes. We measured hematocrit and the concentration of hemoglobin, sTfR, ferritin, EPO, and quantified the effects on performance by measuring time to exhaustion and maximal oxygen uptake (VO2max) on a cycle ergometer. RESULTS Hematocrit increased from 42.7 +/- 1.6% to 50.8 +/- 2.0% in the EPO group, and peaked 1 d after treatment was stopped. In the EPO group, there was an increase in sTfR (from 3.1 +/- 0.9 to 6.3 +/- 2.3 mg x L(-1) , P < 0.001) and in the ratio between sTfR and ferritin (sTfR-ferritin(-1)) (from 3.2 +/- 1.6 to 11.8 +/- 5.1, P < 0.001). The sTfR increase was significant after 1 wk of treatment and remained so for 1 wk posttreatment. Individual values for sTfR throughout the study period showed that 8 of 10 subjects receiving rhEPO, but none receiving placebo, had sTfR levels that exceeded the 95% confidence interval for all subjects at baseline (= 4.6 mg x L(-1)). VO2max increased from 63.6 +/- 4.5 mL x kg(-1) x min(-1) before to 68.1 +/- 5.4 mL x kg(-1) x min(-1) 2 d post rhEPO administration (7% increase, P = 0.001) in the EPO group. Hematocrit, sTfR, sTfR-ferritin(-1), and VO2max did not change in the placebo group. CONCLUSION Serum levels of sTfR may be used as an indirect marker of supranormal erythropoiesis up to 1 wk after the administration of rhEPO, but the effects on endurance performance outlast the increase in sTfR.

Vitamin C and E supplementation hampers cellular adaptation to endurance training in humans: a double-blind, randomised, controlled trial

Recent studies have indicated that antioxidant supplementation may blunt adaptations to exercise, such as mitochondrial biogenesis induced by endurance training. However, studies in humans are sparse and results are conflicting. Isolated vitamin C and E supplements are widely used, and unravelling the interference of these vitamins in cellular and physiological adaptations to exercise is of interest to those who exercise for health purposes and to athletes. Our results show that vitamin C and E supplements blunted the endurance training‐induced increase of mitochondrial proteins (COX4), which is important for improving muscular endurance. Training‐induced increases in V̇O2 max and running performance were not detectably affected by the supplementation. The present study contributes to understanding of how antioxidants may interfere with adaptations to exercise in humans, and the results indicate that high dosages of vitamins C and E should be used with caution.

Subcellular movement and expression of HSP27, αB-crystallin, and HSP70 after two bouts of eccentric exercise in humans

The aims of this study were to investigate the sarcomeric accumulation and expression of heat shock proteins (HSPs) after two bouts of maximal eccentric exercise. Twenty-four subjects performed two bouts of 70 maximal voluntary eccentric actions using the elbow flexors in one arm. The bouts were separated by 3 wk. The changes in concentric (60 degrees/s) and isometric (90 degrees) force-generating capacity were monitored for 9 days after each bout, and biopsies were taken 1 and 48 h and 4 and 7 days after bout 1 and 1 and 48 h after bout 2. The content of HSP27, alphaB-crystallin, HSP70, and desmin in the cytosolic and cytoskeleton/myofibrillar fractions of homogenized muscle samples was determined by immunoassays, and the cellular and subcellular localization of the HSPs in the myofibrillar structure was analyzed by conventional and confocal immunofluorescence microscopy and quantitative electron microscopy. The force-generating capacity was reduced by approximately 50% and did not recover completely during the 3 wk following bout 1. After bout 2, the subjects recovered within 4 days. The HSP levels increased in the cytosolic fraction after bout 1, especially HSP70 (approximately 300% 2-7 days after exercise). Increased levels of HSP27, alphaB-crystallin, and HSP70 were found in the cytoskeletal/myofibrillar fraction after both bouts, despite reduced damage after bout 2. At the ultrastructural level, HSP27 and alphaB-crystallin accumulated in Z-disks, in intermediate desmin-like structures (alphaB-crystallin), and in areas of myofibrillar disruption. In conclusion, HSP27 and alphaB-crystallin accumulated in myofibrillar structures, especially in the Z-disks and the intermediate structures (desmin). The function of the small HSPs is possibly to stabilize and protect the myofibrillar structures during and after unaccustomed eccentric exercise. The large amount of HSP27, alphaB-crystallin, and HSP70 in the cytoskeletal/myofibrillar fraction after a repeated bout of exercise suggests a protective role as part of the repeated-bout effect.

Changes in Calpain Activity, Muscle Structure, and Function after Eccentric Exercise

PURPOSE The aim of this study was to investigate changes in muscle function, muscle structure, and calpain activity after high-force eccentric exercise. METHODS Eleven healthy males performed 300 maximal voluntary eccentric actions with knee extensors in one leg. Maximal force-generating capacity was measured before exercise and regularly during the next 7 d. Biopsies from musculus vastus lateralis were taken in both control and exercised legs 0.5, 4, 8, 24, 96, and 168 h after exercise for evaluation of myofibrillar structure, extracellular matrix proteins, and calpain activity. RESULTS In the exercised leg, peak torque was reduced by 47 +/- 5% during exercise and was still 22 +/- 5% lower than baseline 4 d after the exercise. Calpain activity was three times higher in the exercised leg compared with the control leg 30 min after exercise. Myofibrillar disruptions were observed in 36 +/- 6% of all fibers in exercised muscle and in 2 +/- 1% of fibers in control muscle. The individual reductions in peak torque correlated with the proportion of fibers with myofibrillar disruptions (r = 0.89). The increase in calpain activity was not correlated to the proportion of fibers with myofibrillar disruptions. Nevertheless, the characteristics of the myofibrillar disruptions mimicked calpain-mediated degradation of myofibrils. Tenascin-C and the N-terminal propeptide of procollagen type III showed increased staining intensity on cross-sections 4-7 d after the exercise. CONCLUSIONS Myofibrillar disruptions seem to be a main cause for the long-lasting reduction in force-generating capacity after high-force eccentric exercise. The increase in calpain activity, but the lack of a relationship between calpain activity and the amount of muscle damage, suggests multiple roles of calpain in the damage and repair process.

A COX‐2 inhibitor reduces muscle soreness, but does not influence recovery and adaptation after eccentric exercise

The aim of this study was to investigate the effect of a cyclooxygenase (COX)‐2 inhibitor on the recovery of muscle function, inflammation, regeneration after, and adaptation to, unaccustomed eccentric exercise. Thirty‐three young males and females participated in a double‐blind, placebo‐controlled experiment. Seventy unilateral, voluntary, maximal eccentric actions with the elbow flexors were performed twice (bouts 1 and 2) with the same arm, separated by 3 weeks. The test group participants were administered 400 mg/day of celecoxib for 9 days after bout 1. After both bouts 1 and 2, concentric and isometric force‐generating capacity was immediately reduced (∼40–50%), followed by the later appearance of muscle soreness and increased serum creatine kinase levels. Radiolabelled autologous leukocytes (detected by scintigraphy) and monocytes/macrophages (histology) accumulated in the exercised muscles, simultaneously with increased satellite cell activity. These responses were reduced and recovery was faster after bout 2 than 1, demonstrating a repeated‐bout effect. No differences between the celecoxib and placebo groups were detected, except for muscle soreness, which was attenuated by celecoxib. In summary, celecoxib, a COX‐2 inhibitor, did not detectably affect recovery of muscle function or markers of inflammation and regeneration after unaccustomed eccentric exercise, nor did the drug influence the repeated‐bout effect. However, it alleviated muscle soreness.

Recovery of skeletal muscle contractility after high- and moderate-intensity strength exercise.

Abstract To examine neuromuscular fatigue and recovery, ten male strength athletes [mean (SE) 27.5 (1.4) years] performed a moderate- and a high-intensity strength exercise protocol. In the high-intensity protocol, the load was 100% of the subjects three-repetition maximum (3-RM) for squats and front squats, and 100% of the subjects 6-RM for knee extensions. In the moderate-intensity protocol, the load was 70% of the high-intensity protocol, and both protocols lasted 90 min. The contractile properties of the leg extensor muscles were tested using isokinetic knee extensions, electrical stimulation, and squat jumps. Tests were done before exercise, 5–20 min after exercise, and frequently for 33 h after exercise. The decrements in knee extension performance were greater after the high-intensity protocol (12–14%), as compared to the moderate-intensity protocol (6–7%, P < 0.01). Similar decrements were seen in squat-jumping performance after the high-intensity protocol. Decrements in electrically evoked force were also greatest after the high-intensity protocol (P < 0.05), and were more pronounced at 20 Hz stimulation than at 50 Hz stimulation (P < 0.05). The recovery of performance showed a biphasic pattern, with a rapid recovery within the first 11 h after exercise, followed by a leveling off or a second drop in performance 11–22 h after exercise. All variables were back to baseline by 3 h after the moderate-intensity protocol, while all variables were back to baseline by 33 h after the 100% protocol. The role of structural changes (excitation-contraction coupling and contractile proteins) in the long-lasting performance decrements seen after the high-intensity protocol is discussed.

CHANGES IN HUMAN SKELETAL MUSCLE CONTRACTILITY AND HORMONE STATUS DURING 2 WEEKS OF HEAVY STRENGTH TRAINING

Abstract To examine neuromuscular and hormone changes during 2 weeks of heavy strength training, 18 weight-trained male students were recruited either into a heavy training group (HT, n=11) or into a control group (Ctr, n=7). The heavy training protocol consisted of leg-extensor workouts performed daily, while workouts were performed twice a week in the Ctr group. A test of one repetition maximum (1 RM) was performed before heavy training and on the 2nd day after heavy training. Isokinetic knee extensions, electrical stimulation, and squat jumps were performed before, on the 8th day of heavy training, and on the 4th day after heavy training. Morning blood samples (0800 hours) were drawn before, on the 8th day of heavy training, and on the 4th day after heavy training. Before, and on the 5th day after heavy training, 24 h urine samples were collected. The 1 RM leg press increased by 6 (SEM 2)% in the HT group. Testosterone and insulin-like growth factor-1 concentrations were respectively 12 (SEM 5)% and 11 (SEM 3)% lower than baseline on the 8th day of heavy training; however, hormone levels were back to baseline on the 4th day after heavy training. A significant correlation between individual changes in 1 RM leg press and changes in testosterone concentrations was observed in the HT group (r=0.69). In the HT group, 24 h urinary catecholamine excretion increased by 26 (SEM 12)%, 3-methylhistidine excretion increased by 21 (SEM 6)% and creatinine excretion increased by 11 (SEM 5)%. There were no significant changes in the Ctr group. This work addresses the role of changes in basal hormone status (morning samples) for skeletal muscle adaptation to heavy strength training.

The effect of heavy strength training on muscle mass and physical performance in elite cross country skiers

Aim: To investigate the effect of supplementing high‐volume endurance training with heavy strength training on muscle adaptations and physical performance in elite cross country skiers. Eleven male (18–26 years) and eight female (18–27 years) were assigned to either a strength group (STR) (n=9) or a control group (CON) (n=10). STR performed strength training twice a week for 12 weeks in addition to their normal endurance training. STR improved 1 repetition maximum (RM) for seated pull‐down and half squat (19±2% and 12±2%, respectively), while no change was observed in CON. Cross‐sectional area (CSA) increased in m. triceps brachii for both STR and CON, while there was no change in the m. quadriceps CSA. VO2max during skate‐rollerskiing increased in STR (7±1%), while VO2max during running was unchanged. No change was observed in energy consumption during rollerskiing at submaximal intensities. Double‐poling performance improved more for STR than for CON. Both groups showed a similar improvement in rollerski time‐trial performance. In conclusion, 12 weeks of supplemental heavy strength training improved the strength in leg and upper body muscles, but had little effect on the muscle CSA in thigh muscles. The supplemental strength training improved both VO2max during skate‐rollerskiing and double‐poling performance.