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.

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.

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.

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.

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.

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.

Adequacies of skin puncture for evaluating biochemical and hematological blood parameters in athletes.

OBJECTIVE: The authors tested the effect of blood sampling (skin versus venous puncture) on some biochemical and hematological blood parameters in athletes to answer whether skin puncture could be used as a substitute for venous puncture. DESIGN: Comparative study of 2 methods of blood samples collection. SETTING: The blood was collected in the same athletes at 3 different moments of the preparatory training phase. PARTICIPANTS: Fourteen male indoor soccer players (22 +/- 1 years old) and 7 female handball players (18 +/- 1 years old) participated. MAIN OUTCOME MEASUREMENT: Blood was collected in heparin and K3EDTA by Vacutainer BD or Microvette Sarstedt system for biochemical and hematological analyses, respectively. RESULTS: There were no significant statistical differences between the 2 methods for the values of creatine kinase, urea, creatinine, lymphocytes, and platelets. The other hematological analyzes and uric acid exhibited significant higher values in skin blood, although they were all within the normal expected range. A high degree of correlation was observed between the 2 techniques for all parameters. CONCLUSIONS: Skin puncture is a reliable, easy, accurate, and less invasive sampling method for assessing hematological and some biochemical parameters in athletes, respecting that blood samples should always be obtained from the same site, especially in follow-up studies.