Subcellular fractionation of frozen skeletal muscle samples.

Cell fractionation can be used to determine the localization and trafficking of proteins between cellular compartments such as the cytosol, mitochondria, and nuclei. Subcellular fractionation is usually performed immediately after tissue dissection because freezing may fragment cell membranes and induce organellar cross-contamination. Mitochondria are especially sensitive to freezing/thawing and mechanical homogenization. We proposed a protocol to improve the retention of soluble proteins in the mitochondrial fraction obtained from small amounts of frozen skeletal muscle. Fifty milligrams of the red portion of gastrocnemius muscle from Wistar rats were immediately processed or frozen in liquid nitrogen and stored at -80 °C for further processing. We compared the enrichment of subcellular fractions from frozen/fresh samples obtained with the modified protocol with those obtained by standard fractionation. Western blot analyses of marker proteins for cytosolic (alpha-tubulin), mitochondrial (VDAC1), and nuclear (histone-H3) fractions indicated that all of the procedures resulted in enriched subcellular fractions with minimal organellar cross-contamination. Notably, the activity of the soluble protein citrate synthase was higher in the mitochondrial fractions obtained with the modified protocol from frozen/fresh samples compared with the standard protocol. Therefore, our protocol improved the retention of soluble proteins in the mitochondrial fraction and may be suitable for subcellular fractionation of small amounts of frozen skeletal muscle samples.

Time course of muscle damage and inflammatory responses to resistance training with eccentric overload in trained individuals.

The purpose of this study was to observe the time course of muscle damage and inflammatory responses following an eccentric overload resistance-training (EO) program. 3 females (23.8 ± 2.6 years; 70.9 ± 12.7 kg; 1.6 ± 0.08 m) and 5 males (23.8 ± 2.6 years; 75.1 ± 11.2 kg; 1.8 ± 0.1 m) underwent thirteen training sessions (4 × 8-10 eccentric-only repetitions-80% of eccentric 1RM, one-minute rest, 2x week(-1), during 7 weeks, for three exercises). Blood samples were collected prior to (Pre) and after two (P2), seven (P7), nine (P9), eleven (P11), and thirteen (P13) sessions, always 96 hours after last session. The reference change values (RCV) analysis was employed for comparing the responses, and the percentual differences between the serial results were calculated for each subject and compared with RCV(95%). Four subjects presented significant changes for creatine kinase at P2, and another two at P13; six for C-reactive protein at P2, and three at P11; two for neutrophils at P2, P4, and P13, respectively; and only one for white blood cells at P2, P4, P7, and P9, for lymphocyte at P7, P9, and P13, and for platelet at P4. We conclude that EO induced high magnitude of muscle damage and inflammatory responses in the initial phase of the program with subsequent attenuation.

Proteomic profiling of skeletal muscle in an animal model of overtraining.

Excessive training (i.e. overtraining, OT) may result in underperformance, which can be characterized by the time needed to re-establish performance (i.e. functional overreaching (FOR), nonfunctional overreaching, OT syndrome). The present study is an initial screening for proteins presenting altered abundance in the red (RG) and white (WG) portions of the gastrocnemius muscle from rats submitted to an OT protocol that induced FOR. In the RG, compared to the nontrained control, FOR demonstrated an increased abundance of proteins normally related to adaptation to endurance training (e.g. proteins of oxidative phosphorylation complexes, proteins related to lipid metabolism, antioxidants, and chaperones). In the WG, spots identified as mitochondrial aconitase and a component of the succinate dehydrogenase complex were downregulated in FOR, as were proteins related to myofibril stabilization; these latter were upregulated in the RG. This initial study shows that skeletal muscles with different fiber-type compositions respond differently to an OT period. Also, it is likely that actin-interacting proteins have an important role in muscle adaptation to endurance exercise.

Reproducibility of an incremental treadmill VO(2)max test with gas exchange analysis for runners.

The evaluation of performance through the application of adequate physical tests during a sportive season may be a useful tool to evaluate training adaptations and determine training intensities. For runners, treadmill incremental VO(2)max tests with gas exchange analysis have been widely used to determine maximal and submaximal parameters such as the ventilatory threshold (VT) and respiratory compensation point (RCP) running speed. However, these tests often differ in methodological characteristics (e.g., stage duration, grade, and speed increment size), and few studies have examined the reproducibility of their protocol. Therefore, the aim of this study was to verify the reproducibility and determine the running speeds related to maximal and submaximal parameters of a specific incremental maximum effort treadmill protocol for amateur runners. Eleven amateur male runners underwent 4 repetitions of the protocol (25-second stages, each increasing by 0.3 km·h in running speed while the treadmill grade remained fixed at 1%) after 3 minutes of warm-up at 8-8.5 km·h. We found no significant differences in any of the analyzed parameters, including VT, RCP, and VO(2)max during the 4 repetitions (p > 0.05). Further, the results related to running speed showed high within-subject reproducibility (coefficient of variation < 5.2%). The typical error (TE) values for running speed related to VT (TE = 0.62 km·h), RCP (TE = 0.35 km·h), and VO(2)max (TE = 0.43 km·h) indicated high sensitivity and reproducibility of this protocol. We conclude that this VO(2)max protocol facilitates a clear determination of the running speeds related to VT, RCP, and VO(2)max and has the potential to enable the evaluation of small training effects on maximal and submaximal parameters.

Reference change values of blood analytes from physically active subjects.

The analysis of blood constituents allows the detection of various physiological or pathological states when their values are increased or decreased in relation to a well-defined reference group or to themselves if monitored longitudinally. In the latter case, it is important to know the reference change value (RCV) or critical difference, which defines the percentage change that should be exceeded-given the analytical and biological variations inherent to a particular test, in that there is a significant difference between the two consecutive measurements. Our objective was to calculate the biological variation, analytical variation and RCV of the biochemical and hematological parameters in subjects undergoing 4 months of regular aerobic training. Blood samples (10 mL) were collected monthly from 56 male subjects (17-19 years old). Creatine kinase and aspartate aminotransferase activities, total cholesterol, triglycerides, uric acid, C-reactive protein, alpha-1-acid glycoprotein, creatinine and urea concentrations were measured in sera using an Autolab Boehringer analyzer. Hemogram were obtained from total blood using KX-21 N SYSMEX equipment. The RCV values for leukocytes and all biochemical analytes were elevated compared to the literature values of sedentary subjects. On the other hand, the RCV values for red blood cell count were slightly lower in physically active than in sedentary individuals. Knowledge of analyte RCV values within physically active subjects should improve the sensitivity/specificity of the hematological and biochemical alterations induced by training or the use of recombinant form of erythropoietin through blood parameter analysis, particularly in cases of longitudinal monitoring.

The upper values of plasma creatine kinase of professional soccer players during the Brazilian National Championship.

The current schedule of the Brazilian Soccer Championship may not give players enough recovery time between games. This could increase the chances of muscle damage and impaired performance. We hypothesized that plasma creatine kinase (CK) activity could be a reliable indirect marker of muscle overload in soccer players, so we sought to identify the reference values for upper limits of CK activity during a real-life elite competition. This study analyzed changes in plasma CK activity in 128 professional soccer players at different times during the Brazilian Championship. The upper limits of the 97.5th and 90th percentiles determined for CK activity were 1.338U/L and 975U/L, respectively, markedly higher than values previously reported in the literature. We also evaluated a team monthly throughout the Championship. The upper limit of the 90th percentile, 975U/L, was taken as the decision limit. Six players showing plasma CK values higher than this were asked to decrease their training for 1 week. These players presented lower CK values afterwards. Only one player with a CK value higher than the decision limit (1800U/L 1 day before a game) played on the field and was unfortunately injured during the game. The CK activity in all the other players showed a significant decrease over the course of the Championship, and the values became more homogeneous at the end. The results presented here suggest that plasma CK upper limit values can be used as a practical alternative for early detection of muscle overload in competing soccer players.

Is lactate production related to muscular fatigue? A pedagogical proposition using empirical facts.

The cause-effect relationship between lactic acid, acidosis, and muscle fatigue has been established in the literature. However, current experiments contradict this premise. Here, we describe an experiment developed by first-year university students planned to answer the following questions: 1) Which metabolic pathways of energy metabolism are responsible for meeting the high ATP demand during high-intensity intermittent exercise? 2) Which metabolic pathways are active during the pause, and how do they influence phosphocreatine synthesis? and 3) Is lactate production related to muscular fatigue? Along with these questions, students received a list of materials available for the experiment. In the classroom, they proposed two protocols of eight 30-m sprints at maximum speed, one protocol with pauses of 120 s and the other protocol with pauses of 20 s between sprints. Their performances were analyzed through the velocity registered by photocells. Blood lactate was analyzed before the first sprint and after the eighth sprint. Blood uric acid was analyzed before exercise and 15 and 60 min after exercises. When discussing the data, students concluded that phosphocreatine restoration is time dependent, and this fact influenced the steady level of performance in the protocol with pauses of 120 s compared with the performance decrease noted in the protocol with pauses of 20 s. As the blood lactate levels showed similar absolute increases after both exercises, the students concluded that lactate production is not related to the performance decrement. This activity allows students to integrate the understanding of muscular energy pathways and to reconsider a controversial concept with facts that challenge the universality of the hypothesis relating lactate production to muscular fatigue.

Mountain Ultramarathon Induces Early Increases of Muscle Damage, Inflammation, and Risk for Acute Renal Injury.

Purpose: This study aimed to investigate changes in muscle damage during the course of a 217-km mountain ultramarathon (MUM). In an integrative perspective, inflammatory response and renal function were also studied. Methods: Six male ultra-runners were tested four times: pre-race, at 84 km, at 177 km, and immediately after the race. Blood samples were analyzed for serum muscle enzymes, acute-phase protein, cortisol, and renal function biomarkers. Results: Serum creatine kinase (CK), lactate dehydrogenase (LDH), and aspartate aminotransferase (AST) increased significantly throughout the race (P < 0.001, P < 0.001; P = 0.002, respectively), and effect size (ES) denoted a large magnitude of muscle damage. These enzymes increased from pre-race (132 ± 18, 371 ± 66, and 28 ± 3 U/L, respectively) to 84 km (30, 1.8, and 3.9-fold, respectively); further increased from 84 to 177 km (4.6, 2.9, and 6.1-fold, respectively), followed by a stable phase until the finish line. Regarding the inflammatory response, significant differences were found for C-reactive protein (CRP) (P < 0.001) and cortisol (P < 0.001). CRP increased from pre-race (0.9 ± 0.3 mg/L) to 177 km (243-fold), cortisol increased from pre-race (257 ± 30 mmol/L) to the 84 km (2.9-fold), and both remained augmented until the finish line. Significant changes were observed for creatinine (P = 0.03), urea (P = 0.001), and glomerular filtration rate (GFR) (P < 0.001), and ES confirmed a moderate magnitude of changes in renal function biomarkers. Creatinine and urea increased, and GFR decreased from pre-race (1.00 ± 0.03 mg/dL, 33 ± 6 mg/dL, and 89 ± 5 ml/min/1.73 m2, respectively) to 84 km (1.3, 3.5, and 0.7-fold, respectively), followed by a plateau phase until the finish line. Conclusion: This study shows evidence that muscle damage biomarkers presented early peak levels and they were followed by a plateau phase during the last segment of a 217-km MUM. The acute-phase response had a similar change of muscle damage. In addition, our data showed that our volunteers meet the risk criteria for acute kidney injury from 84 km until they finished the race, without demonstrating any clinical symptomatology.

Hematological and biochemical markers of iron status in a male, young, physically active population.

The aim of this study was to establish reference intervals (RIs) for the hemogram and iron status biomarkers in a physically active population. The study population included male volunteers (n = 150) with an average age of 19 ± 1 years who had participated in a regular and controlled exercise program for four months. Blood samples were collected to determine hematological parameters using a Sysmex XE-5000 analyzer (Sysmex, Kobe, Japan). Iron, total iron-binding capacity (TIBC), transferrin saturation and ferritin, and high-sensitivity C-reactive protein (CRP) concentrations in serum samples were measured using commercial kits (Roche Diagnostics, GmbH, Mannheim, Germany) and a Roche/Hitachi 902 analyzer. The RIs were established using the RefVal program 4.1b. The leucocyte count, TIBC, and CRP and ferritin concentrations exhibited higher RIs compared with those in a nonphysically active population. Thirty volunteers (outliers) were removed from the reference population due to blood abnormalities. Among the outliers, 46% exhibited higher CRP concentrations and lower concentrations of iron and reticulocyte hemoglobin compared with the nonphysically active population (P < 0.001). Our results showed that it is important to establish RIs for certain laboratory parameters in a physically active population, especially for tests related to the inflammatory response and iron metabolism.

Exhaustive exercise with different rest periods changes the collagen content and MMP-2 activation on the calcaneal tendon.

Tendons adapt to different mechanical stimuli through a remodeling process involving metalloproteinases (MMPs) and collagen synthesis. The purpose of this study was to investigate the activities of MMP-2 and MMP-9 and the collagen content in tendons after exhaustive acute exercise sessions over the course of 1, 3, or 6 days, with 1-hr or 3-hr rest periods between each session. Wistar rats were grouped into control (C), trained with 1-hr (groups 1d1h, 3d1h, and 6d1h) and trained with 3-hr (groups 1d3h, 3d3h and 6d3h) groups with rest periods between the treadmill running sessions, for 1, 3, and 6 days. The analysis of MMP-2 showed a larger presence of the latent isoform in the 1d3h group and a larger presence of the active isoform in the 6d3h group compared to the control. No differences were detected for MMP-9. A lower concentration of hydroxyproline was found in the 6d3h group compared to the 6d1h group. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis showed more prominent collagen bands in the 6d3h group, which was confirmed by Western blotting for collagen type I. A higher concentration of glycosaminoglycans was observed in the 3d3h group compared to the 3d1h group, and the 6d3h group presented the highest value for non-collagenous proteins compared to other groups. In conclusion, different rest periods between exercise sessions had different effects on the composition of the calcaneal tendon because a greater activation of MMP-2 and a reduction of total collagen were observed on day 6 of exercise with 3-hr rest periods compared to 1-hr rest periods.

Interaction between overtraining and the interindividual variability may (not) trigger muscle oxidative stress and cardiomyocyte apoptosis in rats.

Severe endurance training (overtraining) may cause underperformance related to muscle oxidative stress and cardiomyocyte alterations. Currently, such relationship has not been empirically established. In this study, Wistar rats (n = 19) underwent eight weeks of daily exercise sessions followed by three overtraining weeks in which the daily frequency of exercise sessions increased. After the 11th training week, eight rats exhibited a reduction of 38% in performance (nonfunctional overreaching group (NFOR)), whereas eleven rats exhibited an increase of 18% in performance (functional overreaching group (FOR)). The red gastrocnemius of NFOR presented significantly lower citrate synthase activity compared to FOR, but similar to that of the control. The activity of mitochondrial complex IV in NFOR was lower than that of the control and FOR. This impaired mitochondrial adaptation in NFOR was associated with increased antioxidant enzyme activities and increased lipid peroxidation (in muscle and plasma) relative to FOR and control. Cardiomyocyte apoptosis was higher in NFOR. Plasma creatine kinase levels were unchanged. We observed that some rats that presented evidence of muscle oxidative stress are also subject to cardiomyocyte apoptosis under endurance overtraining. Blood lipid peroxides may be a suitable biomarker for muscle oxidative stress that is unrelated to severe muscle damage.

Reference intervals for saliva analytes collected by a standardized method in a physically active population.

OBJECTIVES: Our aims were to test a liquid-based saliva collection system for total antioxidant status (TAS), uric acid (UA), total protein concentration (TP) and salivary alpha-amylase (SAA) activity; to determine if these analytes in serum and saliva are correlated in a physically active population and to establish reference intervals for these parameters. DESIGN AND METHODS: Participants in this study were 115 physically active males (18-20 years old). Saliva samples were collected using the Saliva Collection System (Greiner Bio-One) immediately before obtaining blood. Biochemical analyses were conducted using an Autolab Boehringer analyzer. RESULTS: We found a correlation between UA and TP concentrations in serum and saliva samples. The reference intervals for TP and SAA activity in the morning were lower than in the afternoon (p<0.0001). The reference intervals for UA and TAS did not vary with the time of collection. CONCLUSION: The establishment of reference intervals for these saliva constituents increases their diagnostic utility and allows for detection of physiological or pathological states.

The effect of the maintaining the ball possession on the intensity of games

The use of small-sided games (SSG) for soccer training has increased considerably in recent years. Tactical principles can influence the intensity and the movement of soccer players during training. The aim of this study was to evaluate the influence of maintaining possession of the ball on the intensity and the time of motion characteristics of players during training in games. Eleven athletes of the U-20 category of a professional club in São Paulo, Brazil, participated in this study. To check the influence of the tactical principle, we used a game control and an experimental game in three different field dimensions and number of players. The results show that possession of the maintenance rule influences the intensity of the games (p <0.05). However, it is concluded that the rule is fundamental for training through games.

Technical and tactical soccer players' performance in conceptual small-sided games / Desempenho técnico e tático de jogadores de futebol em jogos reduzidos conceituais / Rendimiento técnico y táctico de jugadores de fútbol en juegos reducidos conceptuales

Conceptual small-sided games (CSSGs) may be interesting as a methodology for training soccer players given its connection to the unpredictability that is inherent to soccer. Our aim was investigate, through videogrammetry, if the technical and tactical principles promoted through the adoption of distinct rules from two distinct CSSGs (maintaining ball possession; and progression to the target) would actually be achieved. The study included 24 athletes assigned to 6-player teams. Our data showed that the CSSGs' organising principles create situations with differing levels of difficulty that obey the propositions of maintaining ball possession and progression to the target, i.e., CSSGs permit systematic training on technical and tactical components in order to emphasize the concepts adopted in this study in games context. Our data credit the CSSGs for teaching technical and tactical lessons that, when coupled with adequate physical conditioning, can facilitate a player's capacity to merge thoughts and events in different situations.

A utilização de jogos reduzidos conceituais (JRC) pode ser uma metodologia de treino interessante para treinar a imprevisibilidade inerente ao jogo de futebol. Nosso objetivo no presente estudo foi investigar por videogrametria os princípios técnico-táticos adotados em resposta as respectivas regras de dois jogos reduzidos conceituais distintos (manutenção de posse de bola e progressão ao alvo). Participaram do estudo 24 atletas separados em equipes de 6 jogadores. Nossos dados comprovaram que os princípios organizadores dos JRC criam situações distintas com graus de dificuldade diferentes. Ou seja, é possível sistematizar o treinamento dos componentes técnicos e táticos de forma a enfatizar os conceitos adotados neste estudo e inseridos no contexto do jogo. Nossos dados credenciam os JRC para o aprendizado de situações técnico táticas que, aliadas ao condicionamento físico adequado podem contribuir para produzir um jogador treinado para aproximar cada vez mais pensamentos e ações em situações diversificadas.

El uso de los juegos reducidos conceptuales (JRC) puede ser una interesante metodología para entrenar a la imprevisibilidad inherente en el juego de fútbol. Nuestro objetivo en este estudio fue investigar por videogrametría los principios técnico-tácticos adoptadas en respuesta a las respectivas reglas de dos JRC distintos (posesión del balón y la progresión a la meta). El estudio incluyó a 24 atletas divididos en equipos de 6 jugadores. Nuestros datos muestran que los principios de organización de los JRC crean diferentes situaciones con diferentes grados de dificultad. Es decir, es posible sistematizar el desarrollo de los componentes técnicos y tácticos de una manera a enfatizar los conceptos adoptados para este estudio en el contexto del juego. Nuestros datos acreditan JRC para el aprendizaje de situaciones técnicas tácticas que, junto con la preparación física adecuada puede ayudar a producir jugadores capacitados a unir cada vez más pensamientos y acciones en diferentes situaciones.

Understanding the glycemic index and glycemic load and their practical applications.

We have introduced the study of synthesis pathways using two experiments: 1-the determination of the glycemic index (GI) of some foods and the effects of fiber and fat on the GI; 2-the determination of blood glucose levels after the ingestion of meals with high and low glycemic loads (GL). After a practice assembly, when the foods and meals that were eaten by the students were tallied, the students were divided into groups. At the next class, three members of each group, who had fasted for 8 hr, ingested 50 g of carbohydrate in food or a meal. After ingestion, the blood glucose was measured with a portable device every 30 min for a period of 2 hr. Discussion of the data obtained in experiment 1 allowed the students to understand the mechanism of action of insulin and to understand how the GI, as presented in the literature, is determined. The students also concluded that the addition of fiber to food reduces the glycemic response even with high GI foods, and these results could be a useful strategy for diet prescription. Discussion of experiment 2 allowed the students to understand that the amount of food intake is a determining factor for the glycemic response and subsequent release of insulin. These experimental observations allowed the students to transfer theoretical knowledge to their daily lives very easily. The students approved the classes and felt encouraged to study the synthesis pathways and metabolic integration in the fed state.

Development and characterization of an overtraining animal model.

PURPOSE: Development of an endurance training-overtraining protocol for Wistar rats that includes increased workload and is characterized by analyses of performance and biomarkers. METHODS: The running protocol lasted 11 wk: 8 wk of daily exercise sessions followed by 3 wk of increasing training frequency (two, three, and four times), with decreasing recovery time between sessions (4, 3, and 2 h) to cause an imbalance between overload and recovery. The performance tests were made before training (T1) and after the 4th (T2), 8th (T3), 9th (T4), 10th (T5), and 11th (T6) training weeks. All rats showed significantly increased performance at T4, at which time eight rats, termed the trained group (Tr), were sacrificed for blood and muscle assays. After T6, two groups were distinguishable by differences in the slope (alpha) of a line fitted to the individual performances at T4, T5, and T6: nonfunctional overreaching (NFOR; alpha < -15.05 kg x m) and functional overreaching (FOR; alpha >or= -15.05 kg x m). RESULTS: Data were presented as mean +/- SD. FOR maintained the performance at T6 similar to Tr at T4 (530.6 +/- 85.3 and 487.5 +/- 61.4 kg x m, respectively). The FOR and the Tr groups showed higher muscle citrate synthase activity (approximately 40%) and plasma glutamine/glutamate ratio (Gm/Ga; 4.5 +/- 1.7 and 4.5 +/- 0.9, respectively) than the sedentary control (CO) group (2.8 +/- 0.5). The NFOR group lost the performance acquired at T4 (407.3 +/- 88.2 kg x m) after T6 (280.5 +/- 93.1 kg x m) and exhibited sustained leukocytosis. NFOR's Gm/Ga (3.1 +/- 0.2) and muscle citrate synthase activity were similar to CO values. CONCLUSIONS: The decline in performance in the NFOR group could be related to the decrease in muscle oxidative capacity. We observed a trend in the Gm/Ga and leukocytosis that is similar to what has been sometimes observed in overtrained humans. This controlled training-overtraining animal model may be useful for seeking causative mechanisms of performance decline.

Análise da potência aeróbia de futebolistas por meio de teste de campo e teste laboratorial / Analysis of aerobic power of soccer players through field test and laboratory test / Análisis de la capacidad aeróbica de futbolistas por medio de test de campo y test en laboratório

Introdução: Há métodos diretos e indiretos que são utilizados pelos clubes de futebol para avaliar, acompanhar e determinar o VO2max dos jogadores, sendo este muito importante para o rendimento e a recuperação dos atletas durante uma partida. Objetivo: Avaliar o nível de correlação entre as medidas de VO2max por meio da análise direta de gases e do teste de campo Yo-Yo Intermitente Recovery Level 1 (Yo-YoIR1). Métodos: Participaram do estudo 24 jogadores de futebol da categoria SUB-20 de um clube do estado de São Paulo, Brasil, com estatura 1,72±0,08 m e massa corporal 61,17±9,18 kg, com no mínimo cinco anos de prática na modalidade. Os atletas realizaram o teste de análise direta dos gases em esteira ergométrica e após 48 horas foi realizado o Yo-Yo IR1. Resultados: Os resultados apontaram uma correlação significante entre os testes (r=0,524; p<0,01), porém o Yo-Yo IR1 subestimou as medidas de análise direta dos gases em laboratório (44,98ml/kg/min e 48,14ml/kg/min, respectivamente). Conclusão: Os resultados apontaram uma correlação moderada entre as medidas de VO2max, dessa forma pode-se utilizar ambos os testes para análise da potência aeróbia dos jogadores de futebol, desde que seja repetido o mesmo protocolo nas avaliações subsequentes. .

Introduction: There are direct and indirect methods that are used by soccer clubs to assess, monitor and determine the VO2max of soccer players, which is very important for the performance and recovery of athletes during a match. Objective: The aim of this study was to assess the level of the correlation between measures of VO2max by direct gas analysis and by the field test Yo-Yo Intermittent Recovery Level 1 (Yo-Yo IR1). Methods: The study included 24 Brazilian male soccer players from under 20 category, of a club in the State of São Paulo, Brazil with stature of 1.72 ± 0.08 meter and body mass of 61.17 ± 9.18 kg, with at least five years of practice in the sport. The athletes performed the direct analysis of gases test on a treadmill 48 hours later the Yo-Yo IR1 was performed. Results: The results revealed a significant correlation between tests (r=0.524; p<0.01), but Yo-Yo IR1 underestimated measures of direct analysis of gases in the laboratory (44.98ml/kg/min and 48.14ml/kg/min, respectively). Conclusion: The results showed a moderate correlation between measures of VO2max, that way both tests can be used to analyze the aerobic power of soccer players, since the same protocol is repeated in subsequent evaluations. .

Introducción: Los métodos directos e indirectos son utilizados por los clubes de fútbol para evaluar, monitorear y determinar el VO2max de los jugadores, lo que es muy importante para el rendimiento y la recuperación de los atletas durante un partido. Objetivo: Evaluar el nivel de correlación entre las medidas de VO2max de análisis directo de gases y la prueba de campo Yo-Yo Intermitente Recovery Nivel 1 (Yo-YoIR1). Métodos: El estudio incluyó a 24 futbolistas categoría SUB-20 de un club en el Estado de São Paulo, Brasil, con estatura de 1,72 ± 0,08 metro y masa corporal de 61,17 ± 9,18 Kg, con al menos cinco años de práctica en la modalidad. Los atletas realizaron el análisis directo de los gases en una caminadora y después de 48 horas se realizó el Yo-Yo IR1. Resultados: Los resultados revelaron una correlación significativa entre los ensayos (r = 0,524, p < 0,01), pero el Yo-Yo IR1 ha subestimado las mediciones de laboratorio del análisis directo de gases (44,98 ml/Kg/min. y 48,14 ml/Kg/min., respectivamente). Conclusión: Los resultados indicaron una correlación moderada entre las medidas de VO2max, de esta manera se pueden utilizar ambas las pruebas para analizar la capacidad aeróbica de los jugadores de fútbol, siempre que se repita el mismo protocolo en las evaluaciones posteriores. .

Saliva as a diagnostic fluid in sports medicine: potential and limitations / Saliva como fluido diagnóstico para utilização na medicina esportiva: potencialidades e limitações

The use of saliva in the diagnosis of pathologies and/or monitoring of athletes in competitions or trainings is an attractive alternative due to the fact that samples are easily obtained and it is mostly a less invasive method in comparison with venous blood collection. The saliva is a hypotonic fluid in relation to plasma, containing compounds produced in the salivary glands (immunoglobulin A [IgA] and α-amylase) as well as compounds diffused in the plasma (water, electrolytes, proteins, metabolites and hormones). It plays a pivotal role in the protection of oral mucosa against microbes and in food digestion. Its production and composition depend on the sympathetic and parasympathetic nervous system activity, whose antagonistic action may result in different saliva volumes with distinct ionic and protein profiles. The aim of this review was to present a critical analysis of the potential and limitations of saliva as a diagnostic tool in sports medicine. Although there are studies that have deployed it to monitor athletes in training and doping, the standardization of some preanalytical variables are still required, among which the following ones are worth mentioning: the accurate choice of collection system, which allows the easy quantification of volume with adequate sample recovery; well-defined collection schedules in accordance with the circadian variations of the analyte; prevention of sample contamination with blood from oral mucosa lesions. Another key point for its application in sports is the establishment of reference intervals for analytes quantified in the saliva, collected from a population that comprises healthy individual that exercise regularly and systematically, with physical activity progression.

A utilização de saliva como alternativa para o diagnóstico de patologias e/ou monitoramento de atletas em competições ou treinos é muito atrativa devido à facilidade de obtenção da amostra e, principalmente, pela natureza menos invasiva que a coleta de sangue venoso. A saliva é um fluído hipotônico em relação ao plasma; contém compostos produzidos localmente nas glândulas salivares (imunoglobulina A [IgA] e α-amilase), além de compostos difundidos do plasma (água, eletrólitos, proteínas, metabólitos e hormônios). A saliva desempenha funções importantes na proteção da mucosa oral contra microrganismos e na digestão dos alimentos. Sua produção e sua composição são dependentes da atividade do sistema nervoso autônomo simpático e parassimpático, cuja ação antagônica pode resultar em diferentes volumes de saliva com perfis proteico e iônico distintos. O objetivo da presente revisão é apresentar uma análise crítica das potencialidades e limitações da utilização da saliva como ferramenta diagnóstica para a medicina esportiva. Embora existam estudos que a utilizam para o monitoramento de atletas em situações de exercício e doping, ainda é necessário padronizar algumas variáveis pré-analíticas, como a escolha correta do melhor sistema de coleta, que permite quantificar facilmente o volume, com boa recuperação de amostra; os horários de coleta bem definidos, de acordo com as possíveis variações circadianas do analito; e a contaminação da saliva com sangue proveniente de lesões da mucosa oral, que tem de ser evitada. Outro ponto fundamental para aplicação no esporte é o estabelecimento de valores de referência para analitos quantificados na saliva, obtidos de uma população composta de sujeitos saudáveis e exercitados de forma constante e sistematizada, com progressão de cargas de esforço.

Determination of the anaerobic threshold and maximal lactate steady state speed in equines using the lactate minimum speed protocol.

Maximal blood lactate steady state concentration (MLSS) and anaerobic threshold (AT) have been shown to accurately predict long distance events performance and training loads, as well, in human athletes. Horse endurance races can take up to 160 km and, in practice, coaches use the 4 mM blood lactate concentration, a human based fixed concentration to establish AT, to predict training loads to horse athletes, what can lead to misleading training loads. The lactate minimum speed (LMS) protocol that consists in an initial elevation in blood lactate level by a high intensity bout of exercise and then establishes an individual equilibrium between lactate production and catabolism during progressive submaximal efforts, has been proposed as a nonfixed lactate concentration, to measure individual AT and at the same time predicts MLSS for human long distance runners and basketball players as well. The purpose of this study was to determine the reliability of the LMS protocol in endurance horse athletes. Five male horses that were engaged on endurance training, for at least 1 year of regular training and competition, were used in this study. Animals were submitted to a 500 m full gallop to determine each blood lactate time to peak (LP) after these determinations, animals were submitted to a progressive 1000 m exercise, starting at 15 km h(-1) to determine LMS, and after LMS determination animals were also submitted to two 10,000 m running, first at LMS and then 10% above LMS to test MLSS accuracy. Mean LP was 8.2+/-0.7 mM at approximately 5.8+/-6.09 min, mean LMS was 20.75+/-2.06 km h(-1) and mean heart rate at LMS was 124.8+/-4.7 BPM. Blood lactate remained at rest baseline levels during 10,000 m trial at LMS, but reached a six fold significantly raise during 10% above LMS trial after 4000 and 6000 m (p<0.05) and (p<0.01) after 8000 and 10,000 m. In conclusion, our adapted LMS protocol for horse athletes proposed here seems to be a reliable method to state endurance horse athletes LT and MLSS.

Electrocatalytic determination of reduced glutathione in human erythrocytes.

The determination of reduced glutathione (GSH) in human erythrocytes using a simple, fast and sensitive method employing a glassy carbon electrode modified with cobalt tetrasulfonated phthalocyanine (CoTSPc) immobilized in poly(L: -lysine) (PLL) film was investigated. This modified electrode showed very efficient electrocatalytic activity for anodic oxidation of GSH, decreasing substantially the anodic overpotentials for 0.2 V versus Ag/AgCl. The modified electrode presented better performance in 0.1 mol l(-1) piperazine-N,N'-bis(2-ethanesulfonic acid) buffer at pH 7.4. The other experimental parameters, such as the concentration of CoTSPc and PLL in the membrane preparation, pH, type of buffer solution and applied potential, were optimized. Under optimized operational conditions, a linear response from 50 to 2,160 nmol l(-1) was obtained with a high sensitivity of 1.5 nA l nmol(-1) cm(-2). The detection limit for GSH determination was 15 nmol l(-1). The proposed sensor presented good repeatability, evaluated in terms of the relative standard deviation (1.5%) for n = 10. The modified electrode was applied for determination of GSH in erythrocyte samples and the results were in agreement with those obtained by a comparative method described in the literature The average recovery for these fortified samples was 100 +/- 1)%. Applying a paired Student's-t test to compare these methods, we could observe that, at the 95% confidence level, there was no statistical difference between the reference and the proposed methods.