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Flück, M.1,2, Amami, M.2, van Ginkel, S.2, Dela, F.3, Niederseer, D.3, Niebauer, J.4, Müller, E.4,5 1Balgrist University Hospital (CH), 2Manchester Metropolitan University (UK), 3Xlab University of Copenhagen (De), 4Institute of Sports Medicine of the State Salzburg, 5University of Salzburg (Au) 1 The Institute for Biomedical Research into Human Movement and Health, Manchester Metropolitan University, UK 2Xlab, Department of Biomedical Sciences, Center for Healthy Aging, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark 3 Institute of Sports Medicine of the State of Salzburg, 4 Department of Sport Science and Kinesiology, University of Salzburg, Salzburg, Austria Downhill skiing of the elderly increases maximal oxygen uptake (VO2max) and moderately carbohydrate but not lipid handling (Dela et al 2011). The contribution of skeletal muscle, as a main determinant of activity-induced metabolic adjustments, to improved aerobic fitness with skiing is not understood. We hypothesized that adjustments in aerobic capacity in the main knee extensor muscle, m. vastus lateralis, but not capillary-mediated glucose uptake would explain the improved VO2max with skiing. Methods 28 healthy elderly subjects (age 67.5 ±2.9 years) entered a supervised 3-months downhill ski-training program including an average of 28.5 days of guided skiing. Anthropometric measures were carried out before and after training, including the measure of VO2max, glucose and lipid handling and muscle thickness. Biopsies of vastus lateralis muscle were analysed from 19 subjects (10 males, 9 females) whom completed the training. Cryosections were prepared and capillary density was determined using lectin-based morphometry. Total protein was extracted and subjected to the analysis of the concentration of markers of respiratory chain components (NDUFA9, SDHA, UQCRC1,

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ATP5A1) and the mediator of contraction induced glucose transport, GLUT-4, using SDS-PAGE based immunoblotting with commercially available antibodies. Statistical significance of changes was assessed with a one tailed paired T-test at a p-value of 5%. Results VO2max was increased by 7%, and the HOMA2 index for insulin resistance, but not plasma glucose (p=0.23), decreased by 21% with ski training.

A trend for an increase in the concentration of respiratory chain complexes III (UQCRC1; +60%) and V (ATP5A1, +32%) and capillary density (+12%) was noted. GLUT-4 protein (p=0.49) were not altered. None of measured metabolic proteins correlated with VO2max at either stage of training. Discussion The observations indicate that recreational downhill ski training has a borderline effect on muscle’s aerobic capacity but leaves glucose uptake capacity unaffected. The disconnection between changes in local aerobic capacity and glucose uptake after downhill skiing remains to be explored. References Dela F, Niederseer D, Patsch W, Pirich C, Müller E, Niebauer J. Glucose homeostasis and cardiovascular disease biomarkers in older alpine skiers. Scand J Med Sci Sports 21 S1: 56-61, 2011.

08:30 - 10:00 Oral presentations OP-PM04 Biochemistry [BC] 1 Biochemistry


Hatanaka, E.1, Cury-Boaventura, M.F.1, Gorjão, R.1, de Moura, N.R.1, Santos, V.C.1, Bortolon, J.R.1, Murata, G.M.1,2, Borges, L.S.1, Momesso, C.S.1, Curi, R.2, Pithon-Curi, T.C.1 1ICAFE, Cruzeiro do Sul University (São Paulo, Brazil), 2ICB, University of São Paulo (São Paulo, Brazil) Introduction. After intense exercise, the athlete’s immune response is characterized by suppressed neutrophil function and increased lymphocyte death, resulting in immunosuppression [1,2]. The purpose of this study was to measure necrosis and apoptosis of lymphocytes (DNA fragmentation, surface expression of CD95 and phosphatidylserine) and ROS release in athletes prior to and after a competitive futsal match. Methods. Blood samples were taken from 16 futsal players (19±1 yrs, 68±6 kg, 51±2.6 ml•kg-1•min-1) before and immediately after a futsal match. Lymphocytes were isolated from the players’ peripheral blood using Histopaque®. Lymphocyte necrosis, expression of CD95 and phosphatidylserine, DNA fragmentation and ROS production in lymphocytes were assessed using a flow cytometer. Results. The futsal match induced lymphocytosis and apoptotic death of lymphocytes, as indicated by phosphatidylserine externalization (9.5-fold), CD95 expression (1.5-fold), and DNA fragmentation (1.1-fold). In addition, after the futsal match, lymphocytes spontaneously released higher amounts of ROS (3.6-fold). Discussion. Our results demonstrate that playing futsal induces lymphocyte apoptosis.

During a futsal match, lymphocytes are exposed to pro-apoptotic signals that include augmented levels of inflammatory cytokines and ROS, which elicit changes at the molecular level, leaving lymphocytes more susceptible to both intrinsic and extrinsic pathways for apoptosis. In the extrinsic pathway for apoptosis, the activation of death receptors such as CD95 causes recruitment and oligomerization of the adapter molecule FAS-associated protein with the death domain within the death-inducing signaling complex. Also during apoptosis, membrane asymmetry is lost and phosphatidylserine translocates to the external leaflet of the cell surface, and the cells undergo DNA fragmentation. References. [1] de Moura NR., Cury-Boaventura MF., Santos VC., Levada-Pires AC., Bortolon J., Fiamoncini J., Pithon-Curi TC., Curi R., Hatanaka E. (2012). J Strength Cond Res, 26(9):2507-2514. [2] Walsh NP., Gleeson M., Shephard RJ., Gleeson M., Woods JA., Bishop NC., Fleshner M., Green C., Pedersen BK., Hoffman-Goetz L., Rogers CJ., Northoff H., Abbasi A., Simon P. (2011). Exerc Immunol Rev, 17:6-63.


Lombardi, G.1, Lanteri, P.1, Corsetti, R.2, Grasso, D.1, Di Bernardo, C.1, Colombini, A.1, Graziani, R.3, Banfi, G.1,4 I.R.C.C.S. Istituto Ortopedico Galeazzi 1: IRCCS Istituto Ortopedico Galeazzi (Milano, Italy), 2: Liquigas-Cannondale (Pordenone, Italy), 3: CEDAL (Gallarate, Italy), 4: University of Milano (Milano, Italy) Introduction Cycling stage races are strenuous performances affecting the homeostasis of many tissues, such as bone (Lombardi et al., 2012; Lombardi et al., IN PRESS) and muscles (Colombini et al., 2012). Although explored, the hormonal changes supporting this response is not fully elucidated. Serially measuring the hormonal levels allows a more accurate profiling; this opportunity could be reached with the use of saliva, since salivary steroid hormones are known to reflect type, duration and intensity of exercise (Gatti and De Palo, 2011). Aim of the study was to determine the temporal changes and the reciprocal relationship of salivary steroid hormones, bone-muscular markers and the metabolic effort, during a cycling stage race. Methods Nine pro-cyclists participating in 2012 Giro d’Italia were recruited; daily anthropometrical features and power output/energy expenditure were recorded. Diet was kept constant. Saliva was collected at days -1,4,8, 12, 14, 19, 23; blood and urine were collected at days -1, 12 and 23. Salivary cortisol, DHEA, testosterone and estradiol, serum LDH, AST and CK activities and urinary calcium and phosphorous were measured. Plasma volume changes were determined. Results While cortisol remained constant, testosterone significantly decreased at day 4, estradiol and DHEA increased in the first part of the race to then returned to basal levels. Salivary hormones levels were not correlated with plasma volume shifts. LDH, CK, and AST increased over the race as well as the urinary excretion of calcium an phosphorous. Correlations were found between DHEA and estradiol and the indexes of metabolic effort and the bone-muscular markers. Discussion The present findings on muscular and bone activations are consistent and confirmed those previously published (Lombardi et al., 2012; Lombardi et al., IN PRESS; Colombini et al., 2012). The known co-activation of bone and muscle (Zofkova, 2008) is related to the power output and the energy expenditure and seems to be sustained by the changes in DHEA and estradiol. Saliva represents a useful tool to allow a serial monitoring in the athlete’s hormonal asset even during competitions. We confirm the presence of an hormonal co-regulation of bone and muscle metabolic activities driven by the metabolic effort. References Lombardi G, Lanteri P, Graziani R, et al. (2012). PLoS ONE,7,e42077 Lombardi G, Corsetti R, Lanteri P, et al. Scand J Med Sci Sports IN PRESS Colombini A, Corsetti R, Machado M, et al. (2012) Clin J Sport Med,22,408-413 Gatti R & De Palo EF (2011). Scand J Med Sci Sports,21,157-169 Zofkova I (2008). Physiol Res,57(S1),59-69

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Jamurtas, A., Liakos, Y., Theodorou, A.A., Zalavras, A., Deli, C.K., Fatouros, I.G., Koutedakis, Y.

University of Thessaly Introduction Glucose 6 phosphate dehydrogenase deficiency (G6PD) is the rate limiting step in the pentose phosphate pathway that results in the formation of NADPH which is the driving power for the recycling of glutathione. N-acetylcysteine (NAC) has been used to prevent the decline of glutathione under oxidative stress conditions (Vats et all., 2008) and hypothetically could be efficient under conditions where glutathione levels are reduced such as in G6PD deficiency. Therefore, the purpose of this study was to examine the effects of NAC supplementation on redox status in people with G6PD deficiency. Methods Ten young people (8 males and 2 females) with G6PD deficiency (D) and 10 matched controls with normal levels of the enzyme (N) participated in the study. All subjects received NAC (600 mg per day) for 14 days. Before and after supplementation blood was collected from all participants. Reduced (GSH) and oxidized glutathione (GSSG), thiobarbituric reactive substances (TBARS), bilirubin and uric acid were assessed. Results G6PD levels were 150 times higher in N compared to D (Ν = 9.27 + 1.9 U/g Hb vs. D = 0.62 + 0.55 U/g Hb). There were differences in GSH for group (F1,9= 6.25, p0.05) and time (F1,9= 11.73, p0.005) with N showing significantly higher levels before (Ν = 3.36 + 0.86 μmol/g Hb vs. D = 2.75 + 0.55 μmol/g Hb) and after (Ν = 3.80 + 0.90 μmol/g Hb vs. D = 2.98 + 0.50 μmol/g Hb) supplementation compared to D. No group X time interaction was found for GSH. No significance for group (F1,9= 2.22, p=0.17), time (F1,9= 0.009, p=0.93) or group x time was observed for GSSG levels.

GSH/GSSG ratio was not significant for group (F1,9= 0.19, p=0.67) but approached significance for time (F1,9= 4.205, p=0.07). No significance for group, time or group x time was observed for TBARS, bilirubin or uric acid levels. Discussion These results indicate that short term supplementation with NAC can alter GSH levels in G6PD deficient individuals with no changes in lipid peroxidation or antioxidant indices. Further research is needed in order to examine the effects of longer supplementation with NAC on redox status and the response of these individuals following a stressful condition such as exercise. References Vats P, Singh VK, Singh SN, Singh SB (2008). Glutathione metabolism under high altitude stress and effect of antioxidant supplementation. Aviat Space Environ Med 2008; 79: 1106 – 11.


Kohn, T.A., Noakes, T.D., Rae, D.E., Rubio, J.C., Santalla, A., Lucia, A.

University of Cape Town, Hospital 12 de Octubre, Pablo Olavide University, European University of Madrid Introduction Previous research showed that McArdle patients had a higher surface electrical activity (as measured by electromyography (EMG)) at the same relative workload compared to healthy individuals (2). This suggested that more fibres were activated to fulfil the required work. As muscle fibre type plays an important role in overall muscle function, with type IIa and IIx fibres associated with generating more force than type I fibres, it was hypothesized that McArdle patients would have a predominance of type I myosin heavy chain (MHC) isoforms and less MHC IIa and IIx isoforms compared to healthy individuals. Methods Muscle biopsies were obtained over a 10year period from male and female patients diagnosed with glycogen storage myopathy V (McArdle disease). Nine biopsies from the biceps brachii (BB) (+ 3 healthy controls) and 8 from the vastus lateralis (VL) (+10 healthy controls). These samples were analysed for myosin heavy chain isoform content using SDS-PAGE. The latter provides a relative measure of muscle fibre type. Data (mean ± SD) were analysed using a non-parametric one-way ANOVA. Results In all the samples analysed, all three MHC isoforms were found, namely MHC I, MHC IIa and MHC IIx. No difference in isoform expression was found between McArdle and control VL (MHC I: 33±19 vs. 43±7%;

MHC IIa: 52±9 vs. 40±7%; MHC IIx: 15±18 vs. 17±9%). Similarly, the BB isoform content did not differ significantly between the two groups (MHC I: 33±14 vs. 29±12; MHC IIa: 46±17 vs. 39±5; MHC IIx: 21±13 vs. 32±14%). Discussion This study evaluated whether a difference in muscle fibre type existed between patients diagnosed with McArdle disease and healthy participants. The results showed that the proportion of isoforms expressed in VL and BB are similar to muscle groups from healthy individuals and thus, cannot explain the higher EMG activity recorded. Furthermore, atrophy of muscle fibres from McArdle patients was only reported for type I fibres (1). Therefore, it is speculated that McArdle disease may also affect the contractile apparatus of the skeletal muscle. References 1. Felice KJ, Grunnet ML, Sima AA. Selective atrophy of type 1 muscle fibers in McArdle’s disease. Neurology 47: 581–583, 1996. 2. Rae DE, Noakes TD, San Juan AF, Pérez M, Nogales-Gadea G, Ruiz JR, Moran M, Martín MA, Andreu AL, Arenas J, Lucia A. Excessive skeletal muscle recruitment during strenuous exercise in McArdle patients. Eur. J. Appl. Physiol. 110: 1047–1055, 2010.


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