Revisiting the physical activity paradox: the role of cardiorespiratory fitness in workers with high aerobic demands.

In contrast to leisure time physical activity, occupational physical activity may have adverse health effects-a phenomenon known as the "Physical activity paradox". Characteristics such as long duration, low intensity, static and restricted movement, body position and insufficient recovery are possible explanations as to why physical activity in the occupational context may "wear one out" rather than provide health benefits. We emphasise the role of low cardiorespiratory fitness as a potential contributor to the physical activity paradox, and present data suggesting that only 25% to 50% of Swedish workers in occupations with higher aerobic demands may have "sufficient" cardiorespiratory fitness to maintain good health during their employment. More research is needed to fully understand the complexity of the role of other confounding factors when examining the relationship between cardiorespiratory fitness and occupational workload. However, we believe that there is an increasing need for general awareness amongst Swedish authorities, employees and employers of the potential health consequences of low levels of cardiorespiratory fitness, especially among workers with high occupational workloads. Importantly, when developing interventions targeting the working situation and/or cardiorespiratory fitness levels among workers, researchers should actively involve the relevant population in the design of the study in order to maximize the effect of the interventions on health outcomes.

Reference values for estimated VO2max by two submaximal cycle tests: the Åstrand-test and the Ekblom-Bak test.

AIMS: Submaximal tests estimating VO2max have inherent biases; hence, using VO2max estimations from the same test is essential for reducing this bias. This study aimed to establish sex- and age-specific reference values for estimated VO2max using the Åstrand-test (Å-test) and the Ekblom-Bak test (EB-test). We also assessed the effects of age, exercise level, and BMI on VO2max estimations. METHODS: We included men and women (20-69 years) from the Swedish working population participating in Health Profile Assessments between 2010 and 2020. Excluding those on heart rate-affecting medicines and smokers, n = 263,374 for the Å-test and n = 95,043 for the EB-test were included. VO2max reference values were based on percentiles 10, 25, 40, 60, 75, and 90 for both sexes across 5-year age groups. RESULTS: Estimated absolute and relative VO2max were for men 3.11 L/min and 36.9 mL/min/kg using the Å-test, and 3.58 L/min and 42.4 mL/min/kg using the EB-test. For women, estimated absolute and relative VO2max were 2.48 L/min and 36.6 mL/min/kg using the Å-test, and 2.41 L/min and 35.5 mL/min/kg using the EB-test. Higher age (negative), higher exercise level (positive), and higher BMI (negative) were associated with estimated VO2max using both tests. However, explained variance by exercise on estimated VO2max was low, 10% for the Å-test and 8% for the EB-test, and moderate for BMI, 23% and 29%. CONCLUSION: We present reference values for estimated VO2max from two submaximal cycle tests. Age, exercise, and BMI influenced estimated VO2max. These references can be valuable in clinical evaluations using the same submaximal tests.

Change in cardiorespiratory fitness on self-rated health: prospective cohort study in 98 718 Swedish adults.

Aim: To study how change in cardiorespiratory fitness over time is associated with the development of poor self-rated health in healthy Swedish adults, and whether this association varies with sex, age, body mass index and cardiorespiratory fitness at baseline. A secondary aim was to study the influence of other predictors of self-rated health. Methods: A total of 98,718 participants (45% women, mean age 42.2 years) with two assessments from occupational health service screenings between 1988 and 2019 (mean duration 4.3 years), with good self-rated health at baseline were included. Cardiorespiratory fitness was assessed as estimated maximal oxygen consumption using submaximal cycle testing. Change in cardiorespiratory fitness was expressed as percentage annual change. Poor self-rated health at follow-up was defined as percieving self-rated health as 'poor' or 'very poor'. Results: A large decrease in cardiorespiratory fitness (⩾-3%) was associated with a 34% higher risk of poor self-rated health compared to maintainers (-1 to +1%) after multi-adjustment including change in body mass index, back/neck pain, stress, exercise habits and sleep quality or sleep problems. The associations for decreasers were stronger with longer follow-up time (>10 years). Preserving, or changing to, risk level for body mass index, back/neck pain, stress, exercise and sleep quality/problems were associated with a higher risk of poor self-rated health. Conclusions: Preserving or increasing cardiorespiratory fitness is associated with a lower risk of poor self-rated health, independently of change in other health-related variables, which may act as a protection against future poor self-rated health. This is of high clinical value, and strategies for maintaining or improving cardiorespiratory fitness have the potential to influence both disease and mortality.

Changes in plasma concentration of kynurenine following intake of branched-chain amino acids are not caused by alterations in muscle kynurenine metabolism.

Administration of branched-chain amino acids (BCAA) has been suggested to enhance mitochondrial biogenesis, including levels of PGC-1α, which may, in turn, alter kynurenine metabolism. Ten healthy subjects performed 60 min of dynamic one-leg exercise at ∼70% of Wmax on two occasions. They were in random order supplied either a mixture of BCAA or flavored water (placebo) during the experiment. Blood samples were collected during exercise and recovery, and muscle biopsies were taken from both legs before, after, and 90 and 180 min following exercise. Ingestion of BCAA doubled their concentration in both plasma and muscle while causing a 30%-40% reduction (P < 0.05 vs. placebo) in levels of aromatic amino acids in both resting and exercising muscle during 3-h recovery period. The muscle concentration of kynurenine decreased by 25% (P < 0.05) during recovery, similar in both resting and exercising leg and with both supplements, although plasma concentration of kynurenine during recovery was 10% lower (P < 0.05) when BCAA were ingested. Ingestion of BCAA reduced the plasma concentration of kynurenic acid by 60% (P < 0.01) during exercise and recovery, whereas the level remained unchanged with placebo. Exercise induced a three- to fourfold increase (P < 0.05) in muscle content of PGC-1α1 mRNA after 90 min of recovery under both conditions, whereas levels of KAT4 mRNA and protein were unaffected by exercise or supplement. In conclusion, the reduction of plasma levels of kynurenine and kynurenic acid caused by BCAA were not associated with any changes in the level of muscle kynurenine, suggesting that kynurenine metabolism was altered in tissues other than muscle.

Cardiorespiratory fitness and lifestyle on severe COVID-19 risk in 279,455 adults: a case control study.

BACKGROUND: The impact of cardiorespiratory fitness (CRF) and other lifestyle-related factors on severe COVID-19 risk is understudied. The present study aims to investigate lifestyle-related and socioeconomic factors as possible predictors of COVID-19, with special focus on CRF, and to further study whether these factors may attenuate obesity- and hypertension-related risks, as well as mediate associations between socioeconomic factors and severe COVID-19 risk. METHODS: Out of initially 407,131 participants who participated in nationwide occupational health service screening between 1992 and 2020, n = 857 cases (70% men, mean age 49.9 years) of severe COVID-19 were identified. CRF was estimated using a sub-maximum cycle test, and other lifestyle variables were self-reported. Analyses were performed including both unmatched, n = 278,598, and sex-and age-matched, n = 3426, controls. Severe COVID-19 included hospitalization, intensive care or death due to COVID-19. RESULTS: Patients with more severe COVID-19 had significantly lower CRF, higher BMI, a greater presence of comorbidities and were more often daily smokers. In matched analyses, there was a graded decrease in odds for severe COVID-19 with each ml in CRF (OR = 0.98, 95% CI 0.970 to 0.998), and a two-fold increase in odds between the lowest and highest (< 32 vs. ≥ 46 ml·min-1·kg-1) CRF group. Higher BMI (per unit increase, OR = 1.09, 1.06 to 1.12), larger waist circumference (per cm, OR = 1.04, 1.02 to 1.06), daily smoking (OR = 0.60, 0.41 to 0.89) and high overall stress (OR = 1.36, 1.001 to 1.84) also remained significantly associated with severe COVID-19 risk. Obesity- and blood pressure-related risks were attenuated by adjustment for CRF and lifestyle variables. Mediation through CRF, BMI and smoking accounted for 9% to 54% of the associations between low education, low income and blue collar/low skilled occupations and severe COVID-19 risk. The results were consistent using either matched or unmatched controls. CONCLUSIONS: Both lifestyle-related and socioeconomic factors were associated with risk of severe COVID-19. However, higher CRF attenuated the risk associated with obesity and high blood pressure, and mediated the risk associated with various socioeconomic factors. This emphasises the importance of interventions to maintain or increase CRF in the general population to strengthen the resilience to severe COVID-19, especially in high-risk individuals.

Influence of sex and fiber type on the satellite cell pool in human skeletal muscle.

The repair, remodeling, and regeneration of myofibers are dependent on satellite cells (SCs), although, the distribution of SCs in different fiber types of human muscle remains inconclusive. There is also a paucity of research comparing muscle fiber characteristics in a sex-specific manner. Therefore, the aim of this study was to investigate fiber type-specific SC content in men and women. Muscle biopsies from vastus lateralis were collected from 64 young (mean age 27 ± 5), moderately trained men (n = 34) and women (n = 30). SCs were identified by Pax7-staining together with immunofluorescent analyses of fiber type composition, fiber size, and myonuclei content. In a mixed population, comparable number of SCs was associated to type I and type II fibers (0.07 ± 0.02 vs 0.07 ± 0.02 SCs per fiber, respectively). However, unlike men, women displayed a fiber type-specific distribution, with SC content being lower in type II than type I fibers (P = .041). Sex-based differences were found specifically for type II fibers, where women displayed lower SC content compared to men (P < .001). In addition, positive correlations (r-values between 0.36-0.56) were found between SC content and type I and type II fiber size in men (P = .03 and P < .01, respectively), whereas similar relationships could not be detected in women. Sex-based differences were also noted for fiber type composition and fiber size, but not for myonuclei content. We hereby provide evidence for sex-based differences present at the myocellular level, which may have important implications when studying exercise- and training-induced myogenic responses in skeletal muscle.

Prevalence and time trends of overweight, obesity and severe obesity in 447,925 Swedish adults, 1995-2017.

Aims: The purpose of this research was to describe the current prevalence and historic trends in overweight, obesity and severe obesity in Sweden. Methods: Data on BMI, age, gender, education and geographic region were obtained in n=447,925 Swedish adults through a nationwide screening test (1995-2017). To account for sampling variations, we quantified prevalence estimates and time trends using standardized values (direct method) to all 18-74-year-old Swedes, using nationwide databases. Rates of overweight (BMI ⩾25 kg/m2), obesity (BMI ⩾30 kg/m2) and severe obesity (BMI ⩾35 kg/m2) were calculated across gender, age, education and geographic categories. Years were grouped into two-year sampling periods (except the first period where we used three years) for increased power. We used multivariable logistic regression to quantify independent associations between age, gender, education and region with obesity development and current prevalence rates. Results: In 2016/17 the unstandardized prevalence of overweight, obesity and severe obesity were 55.1%, 16.6% and 4.2%, respectively. Factors associated with a higher obesity prevalence were male gender, older age, lower education and residing in a rural region (all P<0.001). Between 1995 and 2017 the prevalence of severe obesity increased by 153%, compared to obesity (+86%) and overweight (+23%). While there were similar increases in obesity across gender and age groups, people with low education (vs high) and rural areas (vs urban) had a higher prevalence increase (both P<0.001). Conclusions: Rates of overweight, obesity and severe obesity have increased markedly in Swedish adults since 1995. Priority groups for prevention efforts include individuals with low education and those living in rural areas.

Workplace sitting is associated with self-reported general health and back/neck pain: a cross-sectional analysis in 44,978 employees.

BACKGROUND: Total sitting time is associated with a higher risk for cardio metabolic disease and mortality, while breaks in prolonged sitting attenuate these effects. However, less is known about associations of different specific domains and breaks of sitting on general health, back/neck pain and if physical activity could influence these associations. The aim was to investigate how workplace sitting and frequency of breaking up workplace sitting is associated with self-reported general health and self-reported back/neck pain. METHODS: 44,978 participants (42% women) from the Swedish working population, who participated in a nationwide occupational health service screening 2014-2019, were included in this cross-sectional study. Self-reported sitting duration and frequency of breaks from sitting at work, general health, back/neck pain, exercise, leisure time sitting, diet, smoking, stress and body mass index were assessed. Occupation was classified as requiring higher education qualifications or not. Logistic regression modelling was used to assess the association between workplace sitting/frequency of breaks in workplace sitting and poor general health and back/neck pain, respectively. RESULTS: Compared to sitting all the time at work, sitting ≤75% of the time showed significantly lower risks for poor general health (OR range 0.50-0.65), and sitting between 25 and 75% of the time showed significantly lower risks (OR 0.82-0.87) for often reported back/neck pain. For participants reporting sitting half of their working time or more, breaking up workplace sitting occasionally or more often showed significantly lower OR than seldom breaking up workplace sitting; OR ranged 0.40-0.50 for poor health and 0.74-0.81 for back/neck pain. CONCLUSIONS: Sitting almost all the time at work and not taking breaks is associated with an increased risk for self-reported poor general health and back/neck pain. People sitting almost all their time at work are recommended to take breaks from prolonged sitting, exercise regularly and decrease their leisure time sitting to reduce the risk for poor health.

Elevated plasma lactate levels via exogenous lactate infusion do not alter resistance exercise-induced signaling or protein synthesis in human skeletal muscle.

Lactate has been implicated as a potential signaling molecule. In myotubes, lactate incubation increases mechanistic target of rapamycin complex 1 (mTORC1)- and ERK-signaling and induces hypertrophy, indicating that lactate could be a mediator of muscle adaptations to resistance exercise. However, the potential signaling properties of lactate, at rest or with exercise, have not been explored in human tissue. In a crossover design study, 8 men and 8 women performed one-legged resistance exercise while receiving venous infusion of saline or sodium lactate. Blood was sampled repeatedly, and muscle biopsies were collected at rest and at 0, 90, and 180 min and 24 h after exercise. The primary outcomes examined were intracellular signaling, fractional protein synthesis rate (FSR), and blood/muscle levels of lactate and pH. Postexercise blood lactate concentrations were 130% higher in the Lactate trial (3.0 vs. 7.0 mmol/L, P < 0.001), whereas muscle levels were only marginally higher (27 vs. 32 mmol/kg dry wt, P = 0.003) compared with the Saline trial. Postexercise blood pH was higher in the Lactate trial (7.34 vs. 7.44, P < 0.001), with no differences in intramuscular pH. Exercise increased the phosphorylation of mTORS2448 (∼40%), S6K1T389 (∼3-fold), and p44T202/T204 (∼80%) during recovery, without any differences between trials. FSR over the 24-h recovery period did not differ between the Saline (0.067%/h) and Lactate (0.062%/h) trials. This study does not support the hypothesis that blood lactate levels can modulate anabolic signaling in contracted human muscle. Further in vivo research investigating the impact of exercised versus rested muscle and the role of intramuscular lactate is needed to elucidate its potential signaling properties.

Active commuting in Swedish workers between 1998 and 2015-Trends, characteristics, and cardiovascular disease risk.

BACKGROUND: Trend analyses of active commuting and potential variations in trends and association with cardiovascular disease (CVD) risk within subgroups are unknown. OBJECTIVES: To (a) describe trends in active commuting between 1998 and 2015 and (b) to study the association between different amounts of active commuting and the incidence risk of CVD in a large sample of Swedish workers, and analyses of potential variations across subgroups of socio-demographics, physical activity, and BMI. METHODS: A total of 318 309 participants (47% women, 18-74 years) who participated in a nationwide occupational health service screening between 1998 and 2015 were included. Commuting habits were self-reported, and data on first-time CVD events were derived from national registers. RESULTS: Self-reported passive commuters decreased between 1998 and 2015 (64% to 56%), transferring to an increase in mainly moderate/high-dose active commuters (12% to 19%). Changes were seen in all subgroups. The characteristics and lifestyle habits of the typical passive and active commuter changed little over the study period. Low- and moderate/high-dose active commuters had significantly decreased risks for a first time CVD during follow-up. This was accentuated in men, middle-aged, and in participants with light physical work situations, irregular exercise habits, being overweight/obese, and with low fitness. CONCLUSION: Increases in active commuting were observed between 1998 and 2015, however still leaving a majority who do not actively commute. As active commuting, regardless dose, is associated with a lower CVD risk, encouraging more people to actively commute may provide an easily accessible and time-efficient possibility to increase physical activity and health in the general population.

Effects of moderately increased testosterone concentration on physical performance in young women: a double blind, randomised, placebo controlled study.

OBJECTIVE: To investigate the effects of a moderate increase in serum testosterone on physical performance in young, physically active, healthy women. METHODS: A double blind, randomised, placebo controlled trial was conducted between May 2017 and June 2018 (ClinicalTrials.gov ID: NCT03210558). 48 healthy, physically active women aged 18-35 years were randomised to 10 weeks of treatment with 10 mg of testosterone cream daily or placebo (1:1). All participants completed the study. The primary outcome measure was aerobic performance measured by running time to exhaustion (TTE). Secondary outcomes were anaerobic performance (Wingate test) and muscle strength (squat jump (SJ), counter movement jump (CMJ) and knee extension peak torque). Hormone levels were analysed and body composition assessed by dual energy X-ray absorptiometry. RESULTS: Serum levels of testosterone increased from 0.9 (0.4) nmol/L to 4.3 (2.8) nmol/L in the testosterone supplemented group. TTE increased significantly by 21.17 s (8.5%) in the testosterone group compared with the placebo group (mean difference 15.5 s; P=0.045). Wingate average power, which increased by 15.2 W in the testosterone group compared with 3.2 W in the placebo group, was not significantly different between the groups (P=0.084). There were no significant changes in CMJ, SJ and knee extension. Mean change from baseline in total lean mass was 923 g for the testosterone group and 135 g for the placebo group (P=0.040). Mean change in lean mass in the lower limbs was 398 g and 91 g, respectively (P=0.041). CONCLUSION: The study supports a causal effect of testosterone in the increase in aerobic running time as well as lean mass in young, physically active women.

Myosin heavy chain isoform composition in the deltoid and vastus lateralis muscles of elite handball players.

The purpose of the present study was to compare the myosin heavy chain (MHC) isoform composition of the deltoid and vastus lateralis muscles of the dominant and non-dominant limbs in handball players. Eleven male Greek elite handball players (age 22.6 ± 1.9 yrs, training experience 10.6 ± 2.1 yrs, height 184.1 ± 4.1 cm, and weight 81.0 ± 12.5 kg) participated in the study. Four muscle biopsies were obtained from the dominant and non-dominant deltoid and vastus lateralis muscles during the in-season period. The MHC composition was determined using SDS-PAGE. No significant difference was found between the dominant and non-dominant muscles; Deltoid muscle: MHC I [(95%CI = -1.22, 0.33), P = 0.228], MHC ΙΙa [(95%CI = -0.32, 1.59), P = 0.168] and MHC IIx [(95%CI = -1.49, 1.10), P = 0.749]; Vastus lateralis muscle: MHC I [(95%CI = -0.38, 0.63), P = 0.586], MHC ΙΙa [(95%CI = -0.50, 0.65), P = 0.783] and MHC IIx [(95%CI = -1.08, 0.42), P = 0.355]. The findings of the present study indicate that the greater use of the dominant limbs for throwing actions and body movements in handball do not lead to altered MHC isoform composition compared to the non-dominant limbs.

Sex- and age-specific associations between cardiorespiratory fitness, CVD morbidity and all-cause mortality in 266.109 adults.

The aim was to investigate sex- and age-specific associations between cardiorespiratory fitness, all-cause and cause-specific mortality, and cardiovascular disease (CVD) morbidity. 266.109 participants (47% women, 18-74 years) free from CVD, participating in occupational health service screenings in 1995-2015 were included. CRF was assessed as estimated maximal oxygen consumption (estVO2max) using a submaximal cycle test. Incident cases of first-time CVD event and death from any cause were ascertained through national registers. There were 4244 CVD events and 2750 cases of all-cause mortality during mean 7.6 years follow-up. Male gender, higher age and lower estVO2max were associated with higher all-cause mortality and CVD morbidity incidence rates. Risk reductions with increasing estVO2max were present in all age-groups of men and women. No obvious levelling off in risk was identified in the total cohort. However, women and older age-groups showed no further reduction in higher aggregated estVO2max levels. CVD specific mortality was more associated with estVO2max compared to tumor specific mortality. The risk for all-cause mortality and CVD morbidity decreased by 2.3% and 2.6% per increase in 1 ml·min-1·kg-1 with no significant sex-differences but more pronounced in the three lower estVO2max categories for all-cause mortality (9.1%, 3.8% and 3.3%, respectively). High compared to lower levels of estVO2max was not related to a significantly elevated mortality or morbidity. In this large cohort study, CVD morbidity and all-cause mortality were inversely related to estVO2max in both men and women of all age-groups. Increasing cardiorespiratory fitness is a clear public health priority.

Decline in cardiorespiratory fitness in the Swedish working force between 1995 and 2017.

BACKGROUND: Long-term trend analyses of cardiorespiratory fitness (VO2 max) in the general population are limited. OBJECTIVES: To describe trends in VO2 max from 1995 to 2017 in the Swedish working force and to study developments across categories of sex, age, education, and geographic regions. METHODS: A total of 354 277 participants (44% women, 18-74 years) who participated in a nationwide occupational health service screening between 1995 and 2017 were included. Changes in standardized mean values of absolute (L/min) and relative (mL/min/kg) VO2 max, and the proportion with low (<32) relative VO2 max are reported. VO2 max was estimated using a submaximal cycle test. RESULTS: Absolute VO2 max decreased by -6.7% (-0.19 L/min) in the total population. Relative VO2 max decreased by -10.8% (-4.2 mL/min/kg) with approximately one-third explained by a simultaneous increase in body mass. Decreases in absolute fitness were more pronounced in men vs women (8.7% vs 5.3%), in younger vs older (6.5% vs 2.3%), in short (11.4%) vs long (4.5%) education, and in rural vs urban regions (6.5% vs 3.5%), all P < 0.001. The proportions with low VO2 max increased from 27% to 46% (P < 0.001). CONCLUSION: Between 1995 and 2017, there was a steady and pronounced decline in mean cardiorespiratory fitness in Swedish adults. Male gender, young age, short education, and living in a rural area were predictive of greater reductions. The proportion with low cardiorespiratory fitness almost doubled. Given the strong associations between cardiorespiratory fitness and multiple morbidities and mortality, preventing further decreases is a clear public health priority, especially for vulnerable groups.

Correlation of Fiber-Type Composition and Sprint Performance in Youth Soccer Players.

Metaxas, T, Mandroukas, A, Michailidis, Y, Koutlianos, N, Christoulas, K, and Ekblom, B. Correlation of fiber-type composition and sprint performance in youth soccer players. J Strength Cond Res 33(10): 2629-2634, 2019-The aim of this study was to examine the correlation between muscle fiber type and sprint performance in elite young soccer players of different age groups of the same team. Twenty-eight young players participated in this study (group U15, n = 8; group U13, n = 9; and group U11, n = 11). Anthropometric assessments, acceleration (10 m), and Bangsbo modified sprint test (30 m) were performed. Muscle biopsies were obtained from the vastus lateralis, and after that, fiber-type composition was determined by immunohistochemistry. No significant correlations were found between the sprint test and muscle fiber distribution for the groups U13 and U11 (p > 0.05). Also, no correlations were found between cross-sectional areas in the types of fibers with the sprint test in all groups (p > 0.05). A positive correlation was found between type I fibers and the performance in the acceleration test (10 m) (r = 0.77, p < 0.05) was found only in group U15 and a negative correlation between type IIA fibers and the performance in the acceleration test (10 m) (r = -0.89, p < 0.05). The correlations were observed only in group U15, which may indicate that the duration and the intensity of the soccer systematic training can affect the plasticity of the muscle fibers. Specific soccer training in youth is one of the factors that can affect fiber-type plasticity. The specific training programs and status of U15 are more intensive, and the exercises are oriented more to improve physical fitness.

Sex and maturity status affected the validity of a submaximal cycle test in adolescents.

AIM: This study assessed the validity and reliability of the Ekblom-Bak (EB) submaximal cycle test in adolescents and identified any sex- or maturity-related factors for prediction errors. METHODS: We recruited 50 healthy subjects through a public announcement in Stockholm, Sweden, in 2016. The 27 boys and 23 girls were aged 10-15 years and in Tanner stages I-IV. They performed an EB test and incremental treadmill running test for direct measurement of maximal oxygen uptake (VO2 max). RESULTS: The estimation error of VO2 max was 0.09 L/min. The correlation (r) was 0.86, and the standard error of the estimate (SEE) was 0.29 L/min. The largest overestimation was seen in prepubertal boys (0.49 L/min). The best precision of the EB test was achieved when boys in Tanner stages I and II were re-calculated using the prediction equation developed for adult women. This yielded a mean difference of -0.05 L/min, r = 0.92 and SEE 0.23 L/min, in the entire sample. The prediction error was lowered in boys, but not girls, with increasing pubertal maturity. CONCLUSION: The EB test was reasonably valid in adolescents, seemed to be related to sex and maturity status, and our findings support its use.

Activation of mTORC1 by leucine is potentiated by branched-chain amino acids and even more so by essential amino acids following resistance exercise.

Protein synthesis is stimulated by resistance exercise and intake of amino acids, in particular leucine. Moreover, activation of mammalian target of rapamycin complex 1 (mTORC1) signaling by leucine is potentiated by the presence of other essential amino acids (EAA). However, the contribution of the branched-chain amino acids (BCAA) to this effect is yet unknown. Here we compare the stimulatory role of leucine, BCAA, and EAA ingestion on anabolic signaling following exercise. Accordingly, eight trained volunteers completed four sessions of resistance exercise during which they ingested either placebo, leucine, BCAA, or EAA (including the BCAA) in random order. Muscle biopsies were taken at rest, immediately after exercise, and following 90 and 180 min of recovery. Following 90 min of recovery the activity of S6 kinase 1 (S6K1) was greater than at rest in all four trials (Placebo

Control of human energy expenditure by cytochrome c oxidase subunit IV-2.

Resting metabolic rate (RMR) in humans shows pronounced individual variations, but the underlying molecular mechanism remains elusive. Cytochrome c oxidase (COX) plays a key role in control of metabolic rate, and recent studies of the subunit 4 isoform 2 (COX IV-2) indicate involvement in the cellular response to hypoxia and oxidative stress. We evaluated whether the COX subunit IV isoform composition may explain the pronounced individual variations in resting metabolic rate (RMR). RMR was determined in healthy humans by indirect calorimetry and correlated to levels of COX IV-2 and COX IV-1 in vastus lateralis. Overexpression and knock down of the COX IV isoforms were performed in primary myotubes followed by evaluation of the cell respiration and production of reactive oxygen species. Here we show that COX IV-2 protein is constitutively expressed in human skeletal muscle and strongly correlated to RMR. Primary human myotubes overexpressing COX IV-2 displayed markedly (>60%) lower respiration, reduced (>50%) cellular H2O2 production, higher resistance toward both oxidative stress, and severe hypoxia compared with control cells. These results suggest an important role of isoform COX IV-2 in the control of energy expenditure, hypoxic tolerance, and mitochondrial ROS homeostasis in humans.

Intake of branched-chain or essential amino acids attenuates the elevation in muscle levels of PGC-1α4 mRNA caused by resistance exercise.

The transcriptional coactivator peroxisome proliferator-activated receptor-γ coactivator (PGC)-1α is recognized as the master regulator of mitochondrial biogenesis. However, recently a novel isoform, PGC-1α4, that specifically regulates muscle hypertrophy was discovered. Because stimulation of mechanistic target of rapamycin complex 1 (mTORC1) activity is tightly coupled to hypertrophy, we hypothesized that activation of this pathway would upregulate PGC-1α4. Eight male subjects performed heavy resistance exercise (10 × 8-12 repetitions at ∼75% of 1 repetition maximum in leg press) on four different occasions, ingesting in random order a solution containing essential amino acids (EAA), branched-chain amino acids (BCAA), leucine, or flavored water (placebo) during and after the exercise. Biopsies were taken from the vastus lateralis muscle before and immediately after exercise, as well as following 90 and 180 min of recovery. Signaling through mTORC1, as reflected in p70S6 kinase phosphorylation, was stimulated to a greater extent by the EAA and BCAA than the leucine or placebo supplements. Unexpectedly, intake of EAA or BCAA attenuated the stimulatory effect of exercise on PGC-1α4 expression by ∼50% (from a 10- to 5-fold increase with BCAA and EAA, P < 0.05) 3 h after exercise, whereas intake of leucine alone did not reduce this response. The 60% increase (P < 0.05) in the level of PGC-1α1 mRNA 90 min after exercise was uninfluenced by amino acid intake. Muscle glycogen levels were reduced and AMP-activated protein kinase α2 activity and phosphorylation of p38 mitogen-activated protein kinase enhanced to the same extent with all four supplements. In conclusion, induction of PGC-1α4 does not appear to regulate the nutritional (BCAA or EAA)-mediated activation of mTORC1 in human muscle.

Leucine does not affect mechanistic target of rapamycin complex 1 assembly but is required for maximal ribosomal protein s6 kinase 1 activity in human skeletal muscle following resistance exercise.

We examined how the stimulatory effect of leucine on the mechanistic target of rapamycin complex 1 (mTORC1) pathway is affected by the presence of the remaining essential amino acids (EAAs). Nine male subjects performed resistance exercise on 4 occasions and were randomly supplied EAAs with leucine, EAAs without leucine (EAA-Leu), leucine alone, or flavored water (placebo; control). Muscle biopsies were taken from the vastus lateralis before and 60 and 90 min after exercise. Biopsies were analyzed for protein phosphorylation, kinase activity, protein-protein interactions, amino acid concentrations, and tracer incorporation. Leucine alone stimulated ribosomal protein s6 kinase 1 (S6K1) phosphorylation ∼280% more than placebo and EAA-Leu after exercise. Moreover, this response was enhanced by 60-75% after intake of EAAs compared with that of leucine alone (P < 0.05). Kinase activity of S6K1 reflected that of S6K1 phosphorylation; 60 min after exercise, the activity was elevated 3.3- and 4.2-fold with intake of leucine alone and with EAAs, respectively (P < 0.05). The interaction between mammalian target of rapamycin and regulatory-associated protein of mammalian target of rapamycin was unaltered in response to both resistance exercise and amino acid provision. Leucine alone stimulates mTORC1 signaling, although this response is enhanced by other EAAs and does not appear to be caused by alterations in mTORC1 assembly.