Associations between workload, myosin isoforms and performance on professional male basketball. A 4 seasons follow up

The aim of this study was to determine associations between workload, myosin isoforms, and performance in professional basketball, by following the progress of a professional basketball team over four consecutive seasons. Thirty male professional basketball players (age, 27.6 ± 4.1 years;height, 200.1 ± 9.4 cm;weight, 98.5 ± 12.6 kg) from an elite professional basketball team participated in this retrospective observational study. To analyze muscle response and which types of fiber were most involved, fast and myosin in serum were evaluated from three blood samples taken during the season, using enzyme-linked immunosorbent assay (ELISA). Parameters recorded were: exposure time,. Slow and fast myosins for muscle responses. Competitions won, ranking, and mean points scored for performance. Average values per season analysed were 280.1 ± 58 h of exposure to practice,1440.58±533.46µlmol/L of fast and 1178.75±427.75 µmol/L of slow myosin. Performance, assessed as team ranking was 6879.5 ± 985.37 u.a. per season and 90.72±2.79 u.a. per game, winning 7 competitions. Large negative relationships could be observed between slow myosins and exposure time (rho=−0.63;p=.02); There were possible associations between slow myosins and player mean performance per game (R2=0.98;p<.01) and team performance outcomes achieved (R2=0.83;p = 01) during these four seasons. Higher slow serum myosin values could be related to higher exposure time, and lower slow serum myosin values could be associated with better player and team performance. (AU)

Assessment of muscle fiber adaptation in footballers using a new ELISA assay of myosin isoforms.

BACKGROUND: To measure the impact of training models on injury incidence, data of health and performance were integrated to study fiber adaptation during a competitive season. We studied football players over a season, analyzing hours of exposure to sport by serum changes in fast and slow myosin, creatine kinase and lactate dehydrogenase. METHODS: A new assay was developed to measure the myosin isoforms in 49 non-sporting volunteers and in 27 professional football players. RESULTS: Myosin isoforms in volunteers with mean ages of 30±8 were 1553 µg/L fast and 1284 µg/L slow; in the group with of 56±7 were 1426 µg/L fast and 1046 µg/L slow. Slow myosin was significantly lower in older subjects (-18%). Samples from the players in preseason had lower mean scores for fast myosin (1123 µg/L) and higher for slow myosin (2072 µg/L) than reference volunteers. During the season, myosins reached the maximum with the maximum load (1537 µg/L fast, 2195 µg/L slow but decreased and adapted to the high level of demand (425 µg/L fast, 1342 µg/L slow). CK and LDH were maximal at the pre-season (227 U/L, 333 U/L) while myosin levels were maximal at the beginning of season (1537 µg/L, 2195 µg/L). CONCLUSIONS: Measuring serum myosin isoforms we identify the type and amount of damage caused by training and matches, making it a new control tool capable of advising training towards a minimum of blood slow myosin but controlling the fast fiber participating and be able to improve the performance of the players.

Fibre-type-specific and Mitochondrial Biomarkers of Muscle Damage after Mountain Races.

Consequences of running mountain races on muscle damage were investigated by analysing serum muscle enzymes and fibre-type-specific sarcomere proteins. We studied 10 trained amateur and 6 highly trained runners who ran a 35 km and 55 km mountain trail race (MTR), respectively. Levels of creatine kinase (CK), CK-MB isoform (CK-MB), sarcomeric mitochondrial CK (sMtCK), transaminases (AST and ALT), cardiac troponin I (cTnI) and fast (FM) and slow myosin (SM) isoforms, were assessed before, 1 h, 24 h and 48 h after the beginning of MTR. Significant SM increases were found at 24 h in the 55 km group. Levels of CK, CK-MB, AST and cTnI were significantly elevated in both groups following MTR, but in the 55 km group they tended to stabilize in at 48 h. Using pooled data, time-independent serum peaks of SM and CK-MB were significantly correlated. Moreover, concentration of sMtCK was significantly elevated at 1 and 24 h after the race in the 35 km group. Although training volume could confer protection on the mitochondria, the increase in serum CK-MB and SM in the 55 km group might be related to damage to the contractile apparatus type I fibres. Competing in long-distance MTRs might be related to deeper type I muscle fibre damage, even in highly trained individuals.

Early Functional and Morphological Muscle Adaptations During Short-Term Inertial-Squat Training.

Purpose: To assess early changes in muscle function and hypertrophy, measured as increases in muscle cross-sectional areas (CSAs) and total volume, over a 4 weeks inertial resistance training (RT) program. Methods: Ten young RT-naive volunteers (age 23.4 ± 4.1 years) underwent 10 training sessions (2-3 per week) consisting of five sets of 10 flywheel squats (moment of inertia 900 kg⋅cm2). Magnetic resonance imaging (MRI) scans of both thighs were performed before (PRE), and after 2 (IN) and 4 (POST) weeks of training to compute individual muscle volumes and regional CSAs. Scans were performed after ≥96 h of recovery after training sessions, to avoid any influence of acute muscle swelling. PRE and POST regional muscle activation was assessed using muscle functional MRI (mfMRI) scans. Concentric (CON) and eccentric (ECC) squat force and power, as well as maximal voluntary isometric contraction force (MVIC) of knee extensors and flexors, were measured in every training session. Results: Significant quadriceps hypertrophy was detected during (IN: 5.5% ± 1.9%) and after (POST: 8.6% ± 3.6%) the training program. Increases in squat force (CON: 32% ± 15%, ECC: 31 ± 15%) and power (CON: 51% ± 30%, ECC: 48% ± 27%) were observed over the training program. Knee extensor MVIC significantly increased 28% ± 17% after training, but no changes were seen in knee flexor MVIC. No correlation was found between regional muscular activation in the first session and the % of increase in regional CSAs (r = -0.043, P = 0.164). Conclusion: This study reports the earliest onset of whole-muscle hypertrophy documented to date. The process initiates early and continues in response to RT, contributing to initial increases in force. The results call into question the reliability of mfMRI as a tool for predicting the potential hypertrophic effects of a given strengthening exercise.

Time Course and Association of Functional and Biochemical Markers in Severe Semitendinosus Damage Following Intensive Eccentric Leg Curls: Differences between and within Subjects.

Purpose: To investigate the extent and evolution of hamstring muscle damage caused by an intensive bout of eccentric leg curls (ELCs) by (1) assessing the time course and association of different indirect markers of muscle damage such as changes in the force-generating capacity (FGC), functional magnetic resonance (fMRI), and serum muscle enzyme levels and (2) analyzing differences in the degree of hamstring muscle damage between and within subjects (limb-to-limb comparison). Methods: Thirteen male participants performed six sets of 10 repetitions of an ELC with each leg. Before and at regular intervals over 7 days after the exercise, FGC was measured with maximal isometric voluntary contraction (MVC). Serum enzyme levels, fMRI transverse relaxation time (T2) and perceived muscle soreness were also assessed and compared against the FGC. Results: Two groups of subjects were identified according to the extent of hamstring muscle damage based on decreased FGC and increased serum enzyme levels: high responders (n = 10, severe muscle damage) and moderate responders (n = 3, moderate muscle damage). In the high responders, fMRI T2 analysis revealed that the semitendinosus (ST) muscle suffered severe damage in the three regions measured (proximal, middle, and distal). The biceps femoris short head (BFsh) muscle was also damaged and there were significant differences in the FGC within subjects in the high responders. Conclusion: FGC and serum enzyme levels measured in 10 of the subjects from the sample were consistent with severe muscle damage. However, the results showed a wide range of peak MVC reductions, reflecting different degrees of damage between subjects (high and moderate responders). fMRI analysis confirmed that the ST was the hamstring muscle most damaged by ELCs, with uniform T2 changes across all the measured sections of this muscle. During intensive ELCs, the ST muscle could suffer an anomalous recruitment pattern due to fatigue and damage, placing an excessive load on the BFsh and causing it to perform a synergistic compensation that leads to structural damage. Finally, T2 and MVC values did not correlate for the leg with the smaller FGC decrease in the hamstring muscles, suggesting that long-lasting increases in T2 signals after FGC markers have returned to baseline values might indicate an adaptive process rather than damage.

Skeletal muscle signaling, metabolism, and performance during sprint exercise in severe acute hypoxia after the ingestion of antioxidants.

The aim of this study was to determine if reactive oxygen species (ROS) could play a role in blunting Thr172-AMP-activated protein kinase (AMPK)-α phosphorylation in human skeletal muscle after sprint exercise in hypoxia and to elucidate the potential signaling mechanisms responsible for this response. Nine volunteers performed a single 30-s sprint (Wingate test) in two occasions while breathing hypoxic gas ([Formula: see text] = 75 mmHg): one after the ingestion of placebo and another following the intake of antioxidants (α-lipoic acid, vitamin C, and vitamin E), with a randomized double-blind design. Vastus lateralis muscle biopsies were obtained before, immediately after, and 30- and 120-min postsprint. Compared with the control condition, the ingestion of antioxidants resulted in lower plasma carbonylated proteins, attenuated elevation of the AMP-to-ATP molar ratio, and reduced glycolytic rate (P < 0.05) without significant effects on performance or V̇o2 The ingestion of antioxidants did not alter the basal muscle signaling. Thr172-AMPKα and Thr184/187-transforming growth factor-ß-activated kinase 1 (TAK1) phosphorylation were not increased after the sprint regardless of the ingestion of antioxidants. Thr286-CaMKII phosphorylation was increased after the sprint, but this response was blunted by the antioxidants. Ser485-AMPKα1/Ser491-AMPKα2 phosphorylation increased immediately after the sprints coincident with increased Akt phosphorylation. In summary, antioxidants attenuate the glycolytic response to sprint exercise in severe acute hypoxia and modify the muscle signaling response to exercise. Ser485-AMPKα1/Ser491-AMPKα2 phosphorylation, a known mechanism of Thr172-AMPKα phosphorylation inhibition, is increased immediately after sprint exercise in hypoxia, probably by a mechanism independent of ROS.NEW & NOTEWORTHY The glycolytic rate is increased during sprint exercise in severe acute hypoxia. This study showed that the ingestion of antioxidants before sprint exercise in severe acute hypoxia reduced the glycolytic rate and attenuated the increases of the AMP-to-ATP and the reduction of the NAD+-to-NADH.H+ ratios. This resulted in a modified muscle signaling response with a blunted Thr286-CaMKII but similar AMP-activated protein kinase phosphorylation responses in the sprints preceded by the ingestion of antioxidants.

Metabolic adaptations in skeletal muscle after 84 days of bed rest with and without concurrent flywheel resistance exercise.

As metabolic changes in human skeletal muscle after long-term (simulated) spaceflight are not well understood, this study examined the effects of long-term microgravity, with and without concurrent resistance exercise, on skeletal muscle oxidative and glycolytic capacity. Twenty-one men were subjected to 84 days head-down tilt bed rest with (BRE; n = 9) or without (BR; n = 12) concurrent flywheel resistance exercise. Activity and gene expression of glycogen synthase, glycogen phosphorylase (GPh), hexokinase, phosphofructokinase-1 (PFK-1), and citrate synthase (CS), as well as gene expression of succinate dehydrogenase (SDH), vascular endothelial growth factor (VEFG), peroxisome proliferator-activated receptor gamma coactivator-1 (PGC-1α), and myostatin, were analyzed in samples from m. vastus lateralis collected before and after bed rest. Activity and gene expression of enzymes controlling oxidative metabolism (CS, SDH) decreased in BR but were partially maintained in BRE. Activity of enzymes regulating anaerobic glycolysis (GPh, PFK-1) was unchanged in BR. Resistance exercise increased the activity of GPh. PGC-1α and VEGF expression decreased in both BR and BRE. Myostatin increased in BR but decreased in BRE after bed rest. The analyses of these unique samples indicate that long-term microgravity induces marked alterations in the oxidative, but not the glycolytic, energy system. The proposed flywheel resistance exercise was effective in counteracting some of the metabolic alterations triggered by 84-day bed rest. Given the disparity between gene expression vs. enzyme activity in several key metabolic markers, posttranscriptional mechanisms should be explored to fully evaluate metabolic adaptations to long-term microgravity with/without exercise countermeasures in human skeletal muscle.

Sarcomere Disruptions of Slow Fiber Resulting From Mountain Ultramarathon.

OBJECTIVE: To investigate changes after a mountain ultramarathon (MUM) in the serum concentration of fast (FM) and slow (SM) myosin isoforms, which are fiber-type-specific sarcomere proteins. The changes were compared against creatine kinase (CK), a widely used fiber-sarcolemma-damage biomarker, and cardiac troponin I (cTnI), a widely used cardiac biomarker. METHODS: Observational comparison of response in a single group of 8 endurance-trained amateur athletes. Time-related changes in serum levels of CK, cTnI, SM, and FM from competitors were analyzed before, 1 h after the MUM, and 24 and 48 h after the start of the MUM by 1-way ANOVA for repeated measures or Friedman and Wilcoxon tests. Pearson correlation coefficient was employed to examine associations between variables. RESULTS: While SM was significantly (P = .009) increased in serum 24 h after the beginning of the MUM, FM and cTnI did not change significantly. Serum CK activity peak was observed 1 h after the MUM (P = .002). Moreover, serum peaks of CK and SM were highly correlated (r = .884, P = .004). CONCLUSIONS: Since there is evidence of muscle damage after prolonged mountain running, the increase in SM serum concentration after a MUM could be indirect evidence of slow- (type I) fiber-specific sarcomere disruptions.

Rendimiento académico e inserción laboral de los titulados en medicina por la Universitat de Barcelona. Un estudio longitudinal / Academic achievement and work insertion of Barcelona University medical graduates. A longitudinal study

Introducción: El proceso de Bolonia ha provocado cambios en la estructura universitaria dando protagonismo a los conceptos de competencia y calidad. Esta situación planteó el reto de querer mostrar la contribución de la Universitat de Barcelona (UB) en la formación de médicos para la sociedad catalana y la distribución de estos titulados en relación a todo el colectivo de profesionales colegiados en la provincia de Barcelona. Sujetos y métodos: Estudio longitudinal de cuatro promociones de alumnos de la Facultad de Medicina de la UB, que ingresaron entre 1994 y 2001. Para cada cohorte se calcularon estadísticas de rendimiento académico, de formación especializada y de colegiación después de consultar diferentes bases de datos y utilizando el programa informático R. Resultados: El 85-96% de los alumnos que ingresaron en la Facultad de Medicina obtuvieron el título de licenciado en un plazo inferior a siete años. Del total de licenciados, un 83% constaba registrado en el colegio oficial de médicos de la provincia (COMB). Comparadas con la población colegiada de profesionales, estas promociones destacan por una tasa de feminización mayor (tres de cada cuatro) y tasas de extranjería prácticamente nulas. Conclusiones: Los titulados en medicina por la UB demostraron un alto rendimiento de estudio y se insertaron a la profesión en su entorno geográfico

Introduction: The Bologna process has brought about changes in the structure of the Spanish University giving prominence to the concepts of competence and quality. This posed the challenge of showing the contribution of the University of Barcelona (UB) in the training of doctors to Catalan society, and the distribution of these graduates in relation to the entire group of professional associations in the province of Barcelona. Subjects and methods: Longitudinal study of four classes of freshman students of the Faculty of Medicine, UB, who were admitted between 1994 and 2001. For each cohort academic performance, specialized training and licensing statistics were obtained after looking up different databases and using the R software. Results: Between 85% and 96% of students who entered medical school obtained a degree in Medicine in a period shorter than seven years. Of all graduates comprised 83% recorded in medical professional association of the province (COMB). In comparison with the total professional members, these classes highlighted by a greater proportion of women (3 out of 4) and virtually no immigration rates. Conclusions: Graduates in Medicine from the UB demonstrated high performance of their studies and inserted into the profession in its geographical area

Circulating miR-192 and miR-193b are markers of prediabetes and are modulated by an exercise intervention.

CONTEXT: Diabetes is frequently diagnosed late, when the development of complications is almost inevitable, decreasing the quality of life of patients. However, early detection of affected individuals would allow the implementation of timely and effective therapies. OBJECTIVE: Here we set to describe the profile of circulating microRNAs (miRNAs) in prediabetic patients with the intention of identifying novel diagnostic and therapeutic tools. DESIGN: We used real-time RT-PCR to measure the abundance of 176 miRNAs in serum of a cohort of 92 control and prediabetic individuals with either impaired fasting glucose or impaired glucose tolerance, as well as newly diagnosed diabetic patients. We validated the results in a second cohort of control and prediabetic subjects undergoing a therapeutic exercise intervention, as well as in a mouse model of glucose intolerance. RESULTS: We identified two miRNAs, miR-192 and miR-193b, whose abundance is significantly increased in the prediabetic state but not in diabetic patients. Strikingly, these miRNAs are also increased in plasma of glucose-intolerant mice. Moreover, circulating levels of miR-192 and miR-193b return to baseline in both prediabetic humans and glucose-intolerant mice undergoing a therapeutic intervention consisting in chronic exercise, which succeeded in normalizing metabolic parameters. CONCLUSIONS: Our data show that the pattern of circulating miRNAs is modified by defects in glucose metabolism in a similar manner in mice and humans. This circulating miRNA signature for prediabetes could be used as a new diagnostic tool, as well as to monitor response to intervention.

Flywheel resistance exercise to maintain muscle oxidative potential during unloading.

BACKGROUND: As spaceflight compromises skeletal muscle oxidative and aerobic work capacity, this study assessed the efficacy of resistance exercise (RE) to counteract muscle metabolic perturbations induced by 5 wk unilateral lower limb unloading (UL). METHODS: There were 21 men and women (30-56 yr) who were randomly assigned to either UL with (Group, Grp; UL+RE; N = 10) or without (Grp UL; N = 11) concurrent RE. Iso-inertial RE comprised four sets of seven maximal coupled concentric-eccentric knee extensions executed 2-3 times per week. Percutaneous biopsies were obtained from m. vastus lateralis before and after either intervention. Levels of mRNA expression of factors regulating skeletal muscle oxidative capacity i.e., peroxisome proliferator-activated receptor gamma coactivator-1 (PGC-1alpha) and vascular endothelial growth factor (VEGF), and glycolytic capacity, i.e., phosphofructokinase (PFK), glycogen phosphorylase and synthase, hexokinase, and phosphorylase kinase alpha1, were subsequently analyzed. RESULTS: Grp UL showed decreased (36%) PGC-1alpha expression, increased (1.5-fold) PFK expression, and a trend toward decreased VEGF post-intervention. Grp UL+RE showed no changes. DISCUSSION: These results suggest that 5 wk unloading reduces skeletal muscle oxidative capacity and increases glycolytic enzyme activity. More importantly, only 12 bouts of high-force, low-volume resistance exercise attenuated these responses. Thus, the current resistance exercise paradigm emphasizing eccentric overload effectively counteracts unwarranted metabolic alterations induced by 5 wk unloading and may, therefore, aid in maintaining skeletal muscle integrity and endurance, and hence astronaut health and fitness during spaceflight.

Blood ammonia and lactate as markers of muscle metabolites during leg press exercise.

To examine whether blood lactate and ammonia concentrations can be used to estimate the functional state of the muscle contractile machinery with regard to muscle lactate and adenosine triphosphate (ATP) levels during leg press exercise. Thirteen men (age, 34 ± 5 years; 1 repetition maximum leg press strength 199 ± 33 kg) performed either 5 sets of 10 repetitions to failure (5×10RF), or 10 sets of 5 repetitions not to failure (10×5RNF) with the same initial load (10RM) and interset rests (2 minutes) on 2 separate sessions in random order. Capillary blood samples were obtained before and during exercise and recovery. Six subjects underwent vastus lateralis muscle biopsies at rest, before the first set and after the final exercise set. The 5×10RF resulted in a significant and marked decrease in power output (37%), muscle ATP content (24%), and high levels of muscle lactate (25.0 ± 8.1 mmol·kg wet weight), blood lactate (10.3 ± 2.6 mmol·L), and blood ammonia (91.6 ± 40.5 µmol·L). During 10×5RNF no or minimal changes were observed. Significant correlations were found between: (a) blood ammonia and muscle ATP (r = -0.75), (b) changes in peak power output and blood ammonia (r = -0.87) and blood lactate (r = -0.84), and (c) blood and muscle lactate (r = 0.90). Blood lactate and ammonia concentrations can be used as extracellular markers for muscle lactate and ATP contents, respectively. The decline in mechanical power output can be used to indirectly estimate blood ammonia and lactate during leg press exercise.

Critical role for free radicals on sprint exercise-induced CaMKII and AMPKα phosphorylation in human skeletal muscle.

The extremely high energy demand elicited by sprint exercise is satisfied by an increase in O2 consumption combined with a high glycolytic rate, leading to a marked lactate accumulation, increased AMP-to-ATP ratio, and reduced NAD(+)/NADH.H(+) and muscle pH, which are accompanied by marked Thr(172) AMP-activated protein kinase (AMPK)-α phosphorylation during the recovery period by a mechanism not fully understood. To determine the role played by reactive nitrogen and oxygen species (RNOS) on Thr(172)-AMPKα phosphorylation in response to cycling sprint exercise, nine voluntary participants performed a single 30-s sprint (Wingate test) on two occasions: one 2 h after the ingestion of placebo and another after the intake of antioxidants (α-lipoic acid, vitamin C, and vitamin E) in a double-blind design. Vastus lateralis muscle biopsies were obtained before, immediately postsprint, and 30 and 120 min postsprint. Performance and muscle metabolism were similar during both sprints. The NAD(+)-to-NADH.H(+) ratio was similarly reduced (84%) and the AMP-to-ATP ratio was similarly increased (×21-fold) immediately after the sprints. Thr(286) Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) and Thr(172)-AMPKα phosphorylations were increased after the control sprint (with placebo) but not when the sprints were preceded by the ingestion of antioxidants. Ser(485)-AMPKα1/Ser(491)-AMPKα2 phosphorylation, a known inhibitory mechanism of Thr(172)-AMPKα phosphorylation, was increased only with antioxidant ingestion. In conclusion, RNOS play a crucial role in AMPK-mediated signaling after sprint exercise in human skeletal muscle. Antioxidant ingestion 2 h before sprint exercise abrogates the Thr(172)-AMPKα phosphorylation response observed after the ingestion of placebo by reducing CaMKII and increasing Ser(485)-AMPKα1/Ser(491)-AMPKα2 phosphorylation. Sprint performance, muscle metabolism, and AMP-to-ATP and NAD(+)-to-NADH.H(+) ratios are not affected by the acute ingestion of antioxidants.

Hexokinase 2, glycogen synthase and phosphorylase play a key role in muscle glycogen supercompensation.

BACKGROUND: Glycogen-depleting exercise can lead to supercompensation of muscle glycogen stores, but the biochemical mechanisms of this phenomenon are still not completely understood. METHODS: Using chronic low-frequency stimulation (CLFS) as an exercise model, the tibialis anterior muscle of rabbits was stimulated for either 1 or 24 hours, inducing a reduction in glycogen of 90% and 50% respectively. Glycogen recovery was subsequently monitored during 24 hours of rest. RESULTS: In muscles stimulated for 1 hour, glycogen recovered basal levels during the rest period. However, in those stimulated for 24 hours, glycogen was supercompensated and its levels remained 50% higher than basal levels after 6 hours of rest, although the newly synthesized glycogen had fewer branches. This increase in glycogen correlated with an increase in hexokinase-2 expression and activity, a reduction in the glycogen phosphorylase activity ratio and an increase in the glycogen synthase activity ratio, due to dephosphorylation of site 3a, even in the presence of elevated glycogen stores. During supercompensation there was also an increase in 5'-AMP-activated protein kinase phosphorylation, correlating with a stable reduction in ATP and total purine nucleotide levels. CONCLUSIONS: Glycogen supercompensation requires a coordinated chain of events at two levels in the context of decreased cell energy balance: First, an increase in the glucose phosphorylation capacity of the muscle and secondly, control of the enzymes directly involved in the synthesis and degradation of the glycogen molecule. However, supercompensated glycogen has fewer branches.

Energy metabolism during repeated sets of leg press exercise leading to failure or not.

This investigation examined the influence of the number of repetitions per set on power output and muscle metabolism during leg press exercise. Six trained men (age 34 ± 6 yr) randomly performed either 5 sets of 10 repetitions (10REP), or 10 sets of 5 repetitions (5REP) of bilateral leg press exercise, with the same initial load and rest intervals between sets. Muscle biopsies (vastus lateralis) were taken before the first set, and after the first and the final sets. Compared with 5REP, 10REP resulted in a markedly greater decrease (P<0.05) of the power output, muscle PCr and ATP content, and markedly higher (P<0.05) levels of muscle lactate and IMP. Significant correlations (P<0.01) were observed between changes in muscle PCr and muscle lactate (R(2) = 0.46), between changes in muscle PCr and IMP (R(2) = 0.44) as well as between changes in power output and changes in muscle ATP (R(2) = 0.59) and lactate (R(2) = 0.64) levels. Reducing the number of repetitions per set by 50% causes a lower disruption to the energy balance in the muscle. The correlations suggest that the changes in PCr and muscle lactate mainly occur simultaneously during exercise, whereas IMP only accumulates when PCr levels are low. The decrease in ATP stores may contribute to fatigue.

Upregulation of heart PFK-2/FBPase-2 isozyme in skeletal muscle after persistent contraction.

Fructose-2,6-bisphosphate (Fru-2,6-P(2)) is the most potent allosteric activator of liver 6-phosphofructo-1-kinase enzyme, which is crucial for glycolysis. It is present in skeletal muscle but its importance is controversial as a regulator of muscle glycolysis. This study aims to determine the role of Fru-2,6-P(2) in the control of muscle glycolysis during contraction. Muscle contraction was produced by chronic low-frequency stimulation of rabbit tibialis anterior for 24 h, followed by a rest period of 48 h. To determine muscle glycolysis adaptation, we applied a short functional electrostimulation test using the same system of low-frequency stimulation for 1, 3, and 10 s. The variation in concentration of lactate and pyruvate was used to calculate the flux along the glycolysis pathway and the Fru-1,6-P(2)/Fru-6-P ratio permitted to analyze the 6-phosphofructo-1-kinase activation. Fru-2,6-P(2) levels increased over the 24 h of stimulation and remained elevated after the rest period, this being the only metabolite that kept the changes produced by chronic low-frequency stimulation during the rest. During the short functional electrostimulation test, the glycolytic pathway in stimulated and rested muscle was more active than in control muscle, which coincided with higher kinase activity of the 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2/FBPase-2) enzyme. Furthermore, we found a decrease in muscle, liver, and ubiquitous PFK-2/FBPase-2 isoform expression and an increase in heart isoform expression. For the first time, we demonstrate that a persistent increase in Fru-2,6-P(2) produced by a change in PFK-2/FBPase-2 isoform expression may play an important role in the regulation of muscle glycolysis during the first moments of exercise.

Anaerobic energy expenditure and mechanical efficiency during exhaustive leg press exercise.

Information about anaerobic energy production and mechanical efficiency that occurs over time during short-lasting maximal exercise is scarce and controversial. Bilateral leg press is an interesting muscle contraction model to estimate anaerobic energy production and mechanical efficiency during maximal exercise because it largely differs from the models used until now. This study examined the changes in muscle metabolite concentration and power output production during the first and the second half of a set of 10 repetitions to failure (10RM) of bilateral leg press exercise. On two separate days, muscle biopsies were obtained from vastus lateralis prior and immediately after a set of 5 or a set of 10 repetitions. During the second set of 5 repetitions, mean power production decreased by 19% and the average ATP utilisation accounted for by phosphagen decreased from 54% to 19%, whereas ATP utilisation from anaerobic glycolysis increased from 46 to 81%. Changes in contraction time and power output were correlated to the changes in muscle Phosphocreatine (PCr; r = -0.76; P<0.01) and lactate (r = -0.91; P<0.01), respectively, and were accompanied by parallel decreases (P<0.01-0.05) in muscle energy charge (0.6%), muscle ATP/ADP (8%) and ATP/AMP (19%) ratios, as well as by increases in ADP content (7%). The estimated average rate of ATP utilisation from anaerobic sources during the final 5 repetitions fell to 83% whereas total anaerobic ATP production increased by 9% due to a 30% longer average duration of exercise (18.4 ± 4.0 vs 14.2 ± 2.1 s). These data indicate that during a set of 10RM of bilateral leg press exercise there is a decrease in power output which is associated with a decrease in the contribution of PCr and/or an increase in muscle lactate. The higher energy cost per repetition during the second 5 repetitions is suggestive of decreased mechanical efficiency.

Effects of dietary cis and trans unsaturated and saturated fatty acids on the glucose metabolites and enzymes of rats.

The aim of the present study was to examine whether the level of dietary cis fatty acid (cFA), or the isomers (trans or cis) and/or the saturation of the fatty acids at high dietary fat levels altered the intracellular glucose metabolites and certain regulatory enzyme activities in the skeletal muscle and liver of rats. The animals were fed for 30 d on either a recommended control diet (7 % cFA, w/w) or a high-fat diet (20 % fatty acids, w/w). The high-fat diet was enriched with either cFA, trans fatty acid (tFA), a moderate proportion of saturated fatty acid (MSFA), or a high proportion of saturated fatty acid (HSFA). The most striking findings were observed in the gastrocnemius muscle with a HSFA diet. There was a significant increase in glucose-6-phosphate (306 %), glucose-1-phosphate (245 %), fructose-6-phosphate (400 %), fructose-1,6-bisphosphate (86 %), glyceraldehyde-3-phosphate (38 %), pyruvate (341 %), lactate (325 %), citrate (79 %) and the bisphosphorylated sugars as compared with the cFA diet. These changes were paralleled by an increase in muscle triacylglycerol content (49 %) and a decrease in glucose (39 %). In addition, the amount of cFA and the other types of fatty acid (i.e. tFA and MSFA) led to no great differences in glucose metabolism as compared with the respective control group. These data support the hypothesis that glucose changes induced by a HSFA diet are a multifaceted abnormality. Glucose and lactate transport and intracellular glucose metabolism could be the key biochemical defects involved in this detrimental effect on glucose metabolism.

Metabolic effects of ethanol on primary cell cultures of rat skeletal muscle.

Individuals who have consumed alcohol chronically accumulate glycogen in their skeletal muscles. Changes in the energy balance caused by alcohol consumption might lead to alcoholic myopathy. Experimental models used in the past, such as with skeletal muscle biopsy samples of alcohol-dependent individuals or in animal models, do not distinguish between direct effects and indirect effects (i.e., alterations to the nervous or endocrine system) of alcohol. In the current study, we evaluated the direct effect of ethanol on skeletal muscle glycogen concentrations and related glycolytic pathways. We measured the changes in metabolite concentrations and enzyme activities of carbohydrate metabolism in primary cell cultures of rat skeletal muscle exposed to ethanol for two periods. The concentrations of glycolytic metabolites and the activities of several enzymes that regulate glucose and glycogen metabolism were measured. After a short exposure to ethanol (6 h), glucose metabolism slowed. After 48 h of exposure, glycogen accumulation was observed.

Phosphorylation-dependent translocation of glycogen synthase to a novel structure during glycogen resynthesis.

Glycogen metabolism has been the subject of extensive research, but the mechanisms by which it is regulated are still not fully understood. It is well accepted that the rate-limiting enzymes in glycogenesis and glycogenolysis are glycogen synthase (GS) and glycogen phosphorylase (GPh), respectively. Both enzymes are regulated by reversible phosphorylation and by allosteric effectors. However, evidence in the literature indicates that changes in muscle GS and GPh intracellular distribution may constitute a new regulatory mechanism of glycogen metabolism. Already in the 1960s, it was proposed that glycogen was present in dynamic cellular organelles that were termed glycosomas but no such cellular entities have ever been demonstrated. The aim of this study was to characterize muscle GS and GPh intracellular distribution and to identify possible translocation processes of both enzymes. Using in situ stimulation of rabbit tibialis anterior muscle, we show GS and GPh intracellular redistribution at the beginning of glycogen resynthesis after contraction-induced glycogen depletion. We identify a new "player," a new intracellular compartment involved in skeletal muscle glycogen metabolism. They are spherical structures that were not present in basal muscle, and we present evidence that indicate that they are products of actin cytoskeleton remodeling. Furthermore, for the first time, we show a phosphorylation-dependent intracellular distribution of GS. Here, we present evidence of a new regulatory mechanism of skeletal muscle glycogen metabolism based on glycogen enzyme intracellular compartmentalization.