George Tsalis

(1)H NMR-based metabonomic investigation of the effect of two different exercise sessions on the metabolic fingerprint of human urine.

Physical exercise modifies animal metabolism profoundly. Until recently, biochemical investigations related to exercise focused on a small number of biomolecules. In the present study, we used a holistic analytical approach to investigate changes in the human urine metabolome elicited by two exercise sessions differing in the duration of the rest interval between repeated efforts. Twelve men performed three sets of two 80 m maximal runs separated by either 10 s or 1 min of rest. Analysis of pre- and postexercise urine samples by (1)H NMR spectroscopy and subsequent multivariate statistical analysis revealed alterations in the levels of 22 metabolites. Urine samples were safely classified according to exercise protocol even when applying unsupervised methods of statistical analysis. Separation of pre- from postexercise samples was mainly due to lactate, pyruvate, hypoxanthine, compounds of the Krebs cycle, amino acids, and products of branched-chain amino acid (BCAA) catabolism. Separation of the two rest intervals was mainly due to lactate, pyruvate, alanine, compounds of the Krebs cycle, and 2-oxoacids of BCAA, all of which increased more with the shorter interval. Metabonomics provides a powerful methodology to gain insight in metabolic changes induced by specific training protocols and may thus advance our knowledge of exercise biochemistry.

1H NMR Study on the Short- and Long-Term Impact of Two Training Programs of Sprint Running on the Metabolic Fingerprint of Human Serum

Metabonomics is an established strategy in the exploration of the effects of various stimuli on the metabolic fingerprint of biofluids. Here, we present an application of (1)H NMR-based metabonomics on the field of exercise biochemistry. Fourteen men were assigned to either of two training programs, which lasted 8 weeks and involved sets of 80-m maximal runs separated by either 10 s or 1 min of rest. Analysis of pre- and postexercise serum samples, both at the beginning and end of training, by (1)H NMR spectroscopy and subsequent multivariate statistical techniques revealed alterations in the levels of 18 metabolites. Validated O-PLS models could classify the samples in regard to exercise, the separation being mainly due to lactate, pyruvate, alanine, leucine, valine, isoleucine, arginine/lysine, glycoprotein acetyls, and an unidentified metabolite resonating at 8.17 ppm. Samples were also classified safely with respect to training, the separation being mainly due to lactate, pyruvate, methylguanidine, citrate, glucose, valine, taurine, trimethylamine N-oxide, choline-containing compounds, histidines, acetoacetate/acetone, glycoprotein acetyls, and lipids. Samples could not be classified according to the duration of the rest interval between sprints. Our findings underline the power of metabonomics to offer new insights into the short- and long-term impact of exercise on metabolism.

The double burden of obesity and iron deficiency on children and adolescents in Greece: the Healthy Growth Study.

BACKGROUND Some small cohort studies have noted that obesity co-exists with lower serum iron levels. The present study aimed to examine the association between being overweight and iron deficiency (ID) in a large cohort of Greek children and adolescents. METHODS A representative sample of 2492 primary schoolchildren aged 9-13 years old was examined. Anthropometric, biochemical, clinical, dietary intake and physical activity data were collected. RESULTS The prevalence of ID and iron deficiency anaemia (IDA) was higher in obese boys and girls compared to their normal-weight peers (P < 0.05). Serum ferritin was higher in obese compared to normal-weight boys (P = 0.024) and higher in obese compared to normal-weight and overweight girls (P = 0.001). By contrast, a negative association was found between transferrin saturation and adiposity in both boys and girls (P = 0.001 and P = 0.005). Furthermore, obese girls had significantly higher fibre intake than normal-weight girls (P = 0.048) and also overweight and obese boys and girls recorded significantly fewer pedometer steps than their normal-weight peers (P < 0.001). Finally, obesity more than doubled the likelihood of ID in both boys (odds ratio = 2.83; 95% confidence inteval = 1.65-4.85) and girls (odds ratio = 2.03; 95% confidence interval = 1.08-3.81) after controlling for certain lifestyle and clinical indices as potential confounders. CONCLUSIONS The present study shows that obese children and adolescents were at greater risk for ID and IDA than their normal-weight peers. Low grade inflammation induced by excessive adiposity may be a reason for the observed low iron levels. This is also strengthened by the elevated serum ferritin levels, comprising an acute phase protein that is plausibly increased in inflammation.

Muscle metabolism and performance improvement after two training programmes of sprint running differing in rest interval duration

Abstract Repeated-sprint training often involves short sprints separated by inadequate recovery intervals. The effects of interval duration on metabolic and performance parameters are unclear. We compared the effects of two training programmes, differing in rest interval duration, on muscle (vastus lateralis) metabolism and sprint performance. Sixteen men trained three times a week for 8 weeks, each training session comprising 2–3 sets of two 80-m sprints. Sprints were separated by 10 s (n = 8) or 1 min (n = 8). Both training programmes improved performance in the 100-, 200-, and 300-m sprints, but the improvement was greater in the 10-s group during the final 100 m of the 200- and 300-m runs. Independent of interval duration, training mitigated the drop of muscle ATP after two 80-m sprints. The drop in phosphocreatine and the increases in glucose-6-phosphate and fructose-6-phosphate after two 80-m sprints were greater in the 10-s group. In conclusion, training with a limited number of repeated short sprints (≤10 s) may be more effective in improving speed maintenance in 200- and 300-m runs when performed with a 1:1 rather than a 1:6 exercise-to-rest ratio. This may be due to a greater activation of glycolysis caused, in part, by the limited resynthesis of phosphocreatine during the very short rest interval.

Redox, iron, and nutritional status of children during swimming training

Effects of exercise training on important determinants of childrens long-term health, such as redox and iron status, have not been adequately investigated. The aim of the present study was to examine changes in markers of the redox, iron and nutritional status of boy and girl swimmers during a prolonged period of training. 11 boys and 13 girls, aged 10-11 years, were members of a swimming club. They were assessed at the beginning of the training season, at 13 weeks and at 23 weeks through blood sampling and recording of the diet. Reduced glutathione increased at 13 and 23 weeks, whereas oxidised glutathione decreased at 13 weeks, resulting in an increase of the reduced/oxidised glutathione ratio at 13 and 23 weeks. Total antioxidant capacity, catalase, thiobarbituric acid-reactive substances, hemoglobin, transferrin saturation and ferritin did not change significantly. Carbohydrate intake was below 50% of energy and fat intake was above 40% of energy. Intakes of saturated fatty acids and cholesterol were excessive. Iron intake was adequate but intakes of folate, vitamin E, calcium and magnesium did not meet the recommended daily allowances. No significant differences were found between sexes in any of the parameters measured. In conclusion, child swimmers improved the redox status of glutathione during training, although the intake of antioxidant nutrients did not change. The iron status was not impaired by training. Suboptimal intake of several nutrients suggests the need for nutritional monitoring and education of children athletes.

Monitoring the Response of the Human Urinary Metabolome to Brief Maximal Exercise by a Combination of RP-UPLC-MS and 1H NMR Spectroscopy

The delineation of exercise biochemistry by utilizing metabolic fingerprinting has become an established strategy. We present a combined RP-UPLC-MS and (1)H NMR strategy, supplemented by photometric assays, to monitor the response of the human urinary metabolome to short maximal exercise. Seventeen male volunteers performed two identical sprint sessions on separate days, consisting of three 80 m maximal runs. Using univariate and multivariate analyses, we followed the fluctuation of 37 metabolites at 1, 1.5, and 2 h postexercise. 2-Hydroxyisovalerate, 2-hydroxybutyrate, 2-oxoisocaproate, 3-methyl-2-oxovalerate, 3-hydroxyisobutyrate, 2-oxoisovalerate, 3-hydroxybutyrate, 2-hydroxyisobutyrate, alanine, pyruvate, and fumarate increased 1 h postexercise and then returned toward baseline. Lactate and acetate were higher than baseline at 1 and 1.5 h. Hypoxanthine and inosine remained above baseline throughout the postexercise period. Urate decreased at 1 h and increased at 1.5 h before returning to baseline. Valine, isoleucine, succinate, citrate, trimethylamine, trimethylamine N-oxide, tyrosine, and formate decreased at 1 h and/or 1.5 h postexercise and then returned to baseline. Creatinine gradually decreased over the sampling period. Glycine, 4-aminohippurate, and hippurate remained below baseline throughout the postexercise period. Our findings show that even one-half minute of maximal exercise elicited major perturbations in human metabolism, several of which persisted for at least 2 h.

Effects of endurance and high-intensity swimming exercise on the redox status of adolescent male and female swimmers

Abstract Throughout adolescence, swimmers begin to carry out demanding endurance and high-intensity training sets, the effect of which on redox status is largely unknown. The aim of the present study was to investigate the effects of 2000-m continuous swimming and 6 × 50-m maximal swimming on the redox status of adolescent swimmers. Fifteen male and 15 female swimmers, aged 14–18 years, provided blood samples before, immediately after, 1 h after, and 24 h after each exercise for the determination of redox status parameters. Oxidative damage was short-lived and manifest as increases in 8-hydroxy-2΄-deoxyguanosine (8-OHdG) 1 h after high-intensity exercise (39%, P < 0.001) and in malondialdehyde immediately after both exercises (65%, P < 0.001). Alterations in antioxidant parameters were sustained during recovery: reduced glutathione decreased 24 h post-exercise (11%, P = 0.001), uric acid increased gradually after high-intensity exercise (29%, P < 0.001) and bilirubin peaked 24 h post-exercise (29%, P < 0.001). Males had higher 8-OHdG (49%, P = 0.001) and uric acid (29%, P < 0.001) concentrations than females. However, females showed higher values of malondialdehyde than males immediately post-exercise (30%, P = 0.039), despite lower pre-exercise values. In conclusion, both endurance and high-intensity exercise perturbed the redox balance without inducing prolonged oxidative damage in trained adolescent male and female swimmers. These swimming training trials were not found to be detrimental to the redox homeostasis of adolescents.

Biochemical evaluation of running workouts used in training for the 400-m sprint.

Saraslanidis, PJ, Manetzis, CG, Tsalis, GA, Zafeiridis, AS, Mougios, VG, and Kellis, SE. Biochemical evaluation of running workouts used in training for the 400-m sprint. J Strength Cond Res 23(8): 2266-2271, 2009-A 400-m runner relies primarily on the lactate system for energy production. Although several running distances and schemes are used in training for this event, it is not clear which one(s) causes maximal activation of the lactate system so as to optimize adaptations of the lactic capacity. This study examined the effect of 4 running workouts differing in distance (300 vs. 400 m) and mode of execution (continuous/single vs. intermittent) on stimulation of the lactate system and biochemical markers of metabolism and muscle damage. Twelve young men performed 4 runs at maximal effort: 300, 3 × 100, 400, and 2 × 200 m. Blood was drawn before and after exercise for the measurement of lactate, glucose, creatinine, and creatine kinase (CK). Average speed was higher (p < 0.001) in the 300- vs. 400-m tests (7.52 ± 0.50 vs. 7.08 ± 0.59 m·s−1) and in the intermittent vs. continuous tests (7.45 ± 0.50 vs. 7.15 ± 0.58 m·s−1). Lactate was higher in the intermittent vs. continuous tests (16.3 ± 2.2 vs. 15.0 ± 2.0 mmol·L−1, p < 0.05). Serum glucose, creatinine, and CK increased after exercise (p ≤ 0.001), and glucose was higher in the 400- vs. 300-m trials (5.76 ± 0.46 vs. 5.33 ± 0.30 mmol·L−1, p = 0.032). In conclusion, although all 4 running regimens greatly stimulated the lactate system, it appears that the intermittent workouts are superior compared with continuous ones of the same total distance in increasing the ability for energy production via the lactate system. Thus, intermittent workouts can be successfully used by 400-m athletes to develop specific (speed) endurance and should be considered to precede the continuous runs of racing distance within a macrocycle.

Reliability of urine lactate as a novel biomarker of lactate production capacity in maximal swimming

Abstract Context: Postexercise urine lactate may be a novel biomarker of lactate production capacity during exercise. Objective: To evaluate the reliability and utility of the urine lactate concentration after maximal swimming trials between different training protocols (6 × 50 m and 3 × 100 m) and training states (active and nonactive swimmers). Materials and methods: Lactate and creatinine were determined by spectrophotometry in blood and urine. Results: Blood and urine lactate concentrations were correlated in-between training protocols and in participants of different training states. The reliability of the urine lactate concentration was moderate for one of the training protocols and good or moderate for the two training states. Additionally, it was lower than that of the blood lactate concentration, and did not improve after normalizing to the urine creatinine concentration. Discussion and conclusion: Although promising as a biomarker of lactate production capacity, urine lactate requires further research to improve its reliability.

Exercise-induced oxidatively damaged DNA in humans: evaluation in plasma or urine?

Abstract Physical exercise can induce oxidative damage in humans. 8-Hydroxy-2′-deoxyguanosine (8-OHdG) is a widely known biomarker of DNA oxidation, which can be determined in blood and urine. The aim of the present study was to compare these two biological fluids in terms of which is more suitable for the estimation of the oxidative damage of DNA by measuring the concentration of 8-OHdG one hour after maximal exercise by enzyme immunoassay. The concentration of 8-OHdG increased with exercise only in plasma (p < 0.001), and values differed between exercise tests in both plasma and urine (p < 0.05). In conclusion, plasma appears to be more sensitive to exercise-induced 8-OHdG changes than urine and, hence, a more appropriate medium for assessing oxidative damage of DNA, although the poor repeatability of the measurement needs to be addressed in future studies