Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Student: Magdaléna Doleželová Supervisor of master thesis: PharmDr. Miroslav Kovařík, Ph.D. Title of master thesis: Body composition parameters in dependence on the metabolism degrese in patiens with COPD Chronic Obstructive Pulmonary Disease (COPD) is a very serious illness characterized by incompletely reversible airflow obstruction and lung emphysema. Contributes to its overall severity are extrapulmonary manifestations, especially cachexia and loss of lean tissue mass. Chronic inflammation of the respiratory airways and increased respiratory effort cause the hypermetabolic state to some patients with COPD. This thesis investigates the impact of increased metabolism on body composition in patients with COPD. Our study included 50 COPD patients (38 men, 12 women) who were examined by bioelectrical impedance. In this study, we compared the parameters of body composition of men with resting energy expenditure REE > 130% (hereinafter Men over 130%) (n = 9) and the group of men with REE < 130 % (hereinafter Men below 130 %) (n = 29). In the group of Men over 130 % we found a lower average value of body weight by 18 % compared to Men below 130 %. Total body and intracellular water were 12 %...

Une mesure précise de l’activité physique en conditions de vie libre est nécessaire pour une meilleure compréhension de ses relations avec la santé. Le premier objectif de ce travail thèse a été de valider l’Actiheart (qui combine la mesure de la fréquence cardiaque et du mouvement par accélérométrie) par rapport à l’eau doublement marquée (EDM). Nous montrons un bon niveau de concordance entre la dépense énergétique liée à l’activité physique (DEAP) estimée par l’Actiheart et l’EDM. Une individualisation de la relation entre la fréquence cardiaque et de la DEAP par un test d’effort est nécessaire pour une estimation fiable de la DEAP au niveau individuel et pour évaluer des changements de DEAP tels qu’induits par une intervention. En laboratoire, nous montrons que la précision de l’Actiheart est activité dépendante. Ceci nécessite la mise en place de leur reconnaissance par de nouveaux capteurs et modèles mathématiques. L’adiponectine est une hormone du tissu adipeux qui a un rôle dans le métabolisme énergétique et dont la sécrétion diminue avec l’obésité. Les effets de l’activité physique sont en revanche contradictoires dans la littérature. Le second objectif de ce travail a été d’évaluer l’effet de l’activité physique et d’une intervention avec contrôle du poids sur les taux plasmatiques d’adiponectine. Nous montrons que l’adiponectine totale et à haut poids moléculaire sont associées négativement à la variation du niveau d’activité physique. D’autres travaux sont cependant nécessaires pour comprendre les mécanismes qui sous-tendent cette modulation de l’adiponectine plasmatique qui ne semble pas liée à des variations de synthèse dans le tissu adipeux ou musculaire. / Accurate measurements of physical activity in free living are needed to establish what dose of physical activity is necessary for obtaining a specific health benefits. The first aim of this work was to validate the Actiheart (which combines heart rate and accelerometry sensors) with doubly labeled water (DLW). We show a good level of concordance between physical activity energy expenditure (PAEE) estimated by Actiheart and DLW. Individualization of the relationship between heart rate and PAEE by an incremental test is needed for an accurate estimate of the PAEE at the individual level and to evaluate changes induced by an intervention. In laboratory, we show that the accuracy of Actiheart is activitydependent. This requires the establishment of their recognition from new sensors and mathematical models. Adiponectin, hormone secreted by adipose tissue, has a role in energy metabolism and its secretion decreases with obesity. However the effects of physical activity remain in contradiction in published studies. The second objective of this work was to evaluate the effect of physical activity and intervention with weight control on plasma adiponectin. We show that the total and high molecular weight adiponectin were negatively associated with modifications of the physical activity level. Further work is however necessary to understand the mechanisms underlying this modulation of plasma adiponectin which does not seem related to changes in synthesis in adipose tissue or muscle.

Choque séptico é caracterizado por desequilíbrio entre o transporte e o consumo de oxigênio, podendo acarretar hipóxia tecidual. A disfunção microcirculatória, característica cardinal da fisiopatologia do choque séptico, causa má distribuição de fluxo sanguíneo microvascular e, consequentemente, shunt de oxigênio, disóxia tissular e, teoricamente, diminuição no consumo de oxigênio (VO2) pela célula. No presente estudo, foi investigada a associação entre alterações microcirculatórias causadas pela sepse e o consumo de oxigênio em pacientes pediátricos. Dezessete crianças com choque séptico ressuscitadas foram estudadas em quatro momentos durante a internação na unidade de terapia intensiva (dentro de 24, 48 e 72 horas após a admissão ou diagnóstico de choque e após a resolução deste, antes da extubação traqueal). A microcirculação sublingual foi avaliada utilizando o método de imagem Sidestream dark field (SDF) e o VO2 foi calculado através da calorimetria indireta. Outras variáveis hemodinâmicas, como transporte de oxigênio, índice cardíaco, pressão arterial invasiva, lactato arterial e saturação venosa central, foram coletadas. Embora as variáveis hemodinâmicas tenham se mantido em níveis satisfatórios, graves alterações na microcirculação foram visualizadas, especialmente na densidade de vasos pequenos perfundidos (DVPP), na proporção de vasos pequenos perfundidos (PVPP) e no índice de fluxo microvascular (MFI). Foram encontradas assosciações significativas entre o VO2 e os parâmetros da microcirculação: dVO2 e dDVPP (&#946; coefficient= 6,875; p<0,001), dVO2 e dPVPP (&#946; coefficient=92,246; p<0,001) e dVO2 e dMFI (&#946; coefficient=21,213; p<0,001). Não foram encontradas correlações entre as alterações microcirculatórias e as outras variáveis. Em conclusão, este estudo mostrou que pacientes pediátricos com choque séptico apresentaram grave disfunção microvascular e que o fluxo microcirculatório alterado estava associado ao VO2, podendo estar implicado na fisiopatologia da disóxia tecidual da sepse. / Septic shock is characterized by the imbalance between oxygen delivery and consumption leading to tissue hypoxia. Microcirculatory dysfunction, a key element of septic shock pathogenesis, elicits maldistribution of microvascular blood flow and consenquently oxygen shunt, tissue oxygenation debt and, theoretically, impaired oxygen consumption (VO2). In this study, it was investigated if there is an association between microcirculatory changes and VO2 in pediatric patients with septic shock. Seventeen resuscitated patients with septic shock were studied in four moments (within 24hr, 48hr and 72hr of the admission or diagnosis of shock and after its resolution, prior to extubation). Sublingual microcirculation was evaluated using Sidestream dark field (SDF) imaging and VO2 was measured directly by indirect calorimetry. Other hemodynamic variables, like cardiac index, oxygen delivery, invasive arterial pressure, arterial lactate and central venous oxygen saturation were also recorded. Although global hemodynamic variables were within satisfactory ranges, microvascular variables were markedly altered, especially microvascular flow index (MFI), proportion of perfused small vessels (PPV) and perfused small vessel density (PVD). Significant associations between oxygen consumption and microcirculatory parameters were found: dVO2 and dPVD (&#946; coefficient= 6.875; p<0.001), dVO2 and dPPV (&#946; coefficient=92.246; p<0.001) and dVO2 and dMFI (&#946; coefficient=21.213; p<0.001). There was no correlation between microcirculatory alterations and other variables in this study. In conclusion, this study showed that pediatric patients with septic shock presented severe microcirculatory dysfunction and abnormal microvascular blood flow could be associated to oxygen consumption.

L’augmentation de la dépense énergétique (DE) par une augmentation de l'activité physique (AP) participe au maintien et à l’amélioration de la santé. La mesure à grande échelle de la DE totale (DET) en général et AP en particulier se heurte à des difficultés pratiques de recueil de données, de validité et aux coûts. Bien que dans la littérature de nombreux accéléromètres permettent d’estimer la DET, il y a encore des limites quant à la mesure de l’intensité de l’AP, élément qui influence l’état de la balance énergétique et le bénéfice pour la santé de l’AP. De plus, peu de comparaisons entre les différents accéléromètres sont disponibles. La présente étude avait pour but d’évaluer la capacité de deux accéléromètres (Actical et Sensewear Armband) pour estimer la DET en comparaison avec la technique de l’eau doublement marquée (EDM) ; d’évaluer la concordance entre les accéléromètres dans la mesure de la DE au repos (DER) en comparaison avec la technique de la calorimétrie indirecte (CI) et d’évaluer la DE liée à l’AP en comparaison avec la technique de la CI. Les résultats montrent qu’il y a une très bonne corrélation entre la CI et les accéléromètres dans la mesure de la DER(r > 0.80, p<0.001) et une bonne corrélation entre les accéléromètres et l’EDM dans la mesure de la DET(r>0.75, p<0.001). Pour la mesure de l’intensité de l’AP, l’ACT est plus précis (r=0.80, p<0.001)) que le SWA à la marche, le SWA est meilleur(r=0.80, p<0.001) au vélo. Cette étude permet d’affirmer que le SWA et l’ACT sont des alternatives valables pour mesurer la DE. / The increase in energy expenditure by the increase of physical activity contributes to maintaining and improving health. The large-scale measurement of total energy expenditure in general and physical activity in particular, is faced with difficulties of data collection, validity and cost. Although literature suggests that many accelerometers are used to estimate total energy expenditure, there are still limits as to the measurement of the intensity of physical activity, which is one of the factors that influences the energy balance and health benefits of physical activity. In addition, few comparisons between the different accelerometers are available. This study aimed to evaluate the ability of two accelerometers (Actical and SensWear Armband) to estimate total energy expenditure compared with the technique of doubly labelled water; to assess the correlation between the two accelerometers in the measurement of resting energy expenditure, compared with the technique of indirect calorimetry and finally to assess energy expenditure linked to physical (walking and cycling) at different intensities, in comparison with the indirect calorimetry technique. The results show a very good correlation between indirect calorimetry and the two accelerometers(r > 0.80, p<0.001) in the measurement of resting energy expenditure and a good correlation between accelerometers and the gold standard technique (doubly labeled water) in the measurement of total energy expenditure(r>0.75, p<0.001). To measure the intensity of physical activity, Actical is more accurate than SensWear Armband(r=0.80, p<0.001) at low intensity while the SensWear Armband is better at higher intensity (r=0.80, p<0.001). This study provides evidence that these two accelerometers are suitable alternatives for measuring energy expenditure.

Made available in DSpace on 2016-08-15T20:31:12Z (GMT). No. of bitstreams: 1 HericaGTD_DISSERT.pdf: 597617 bytes, checksum: 59784d911ca59573a589df402958dde5 (MD5) Previous issue date: 2012-05-21 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / This study aimed to assess the daily variation of metabolic heat production in goats based on oxygen consumption and carbon dioxide production in association with meteorological variables and physiological responses. 10 SPRD goats were used, four with predominantly white fur and six with predominantly black. The observations were made with the protected and exposed animals to direct sunlight, in a period of 12 days during the months of August and September, 2010 in Mossoró-RN. The metabolic heat production (M, Wm-2), the latent heat flow from the respiratory system (ER, Wm-2) and from the cutaneous surface (ESC Wm-2) were determined simultaneously through a face mask and a ventilated capsule, respectively. We also measured the rectal temperature, surface temperature and respiratory rate and measured the environmental variables, air temperature, black globe temperature, wind speed, air humidity, partial vapor pressure and solar radiation. Analysis of variance was based on least-squares method and comparison of means was done by Tukey's test at 5% level of significance. The results showed that the semi-arid goats maintained stable to metabolic heat production during the day, however, when exposed to direct sunlight, significantly increased the latent heat loss in the cutaneous surface and reduced metabolic heat production approximately 24% in an attempt to maintain constant internal temperature / Este estudo objetivou avaliar a variação diária da produção de calor metabólico em caprinos baseada no consumo de oxigênio e produção de dióxido de carbono em associação com variáveis meteorológicas e respostas fisiológicas. Foram utilizados 10 caprinos SPRD, sendo quatro com pelame predominantemente branco e seis com pelame predominantemente preto. As observações foram feitas com os animais protegidos e expostos à radiação solar direta, em um período de 12 dias, durante os meses de agosto e setembro de 2012, em Mossoró-RN. A produção de calor metabólico (M, Wm-2), o fluxo de calor latente a partir do sistema respiratório (ER, Wm-2 ) e a partir da superfície cutânea (ESC, Wm-2) foram determinados, ao mesmo tempo, por meio de uma máscara facial e uma cápsula ventilada, respectivamente. Foram também aferidas a temperatura retal, temperatura de superfície e a freqüência respiratória e medidas as variáveis ambientais, temperatura do ar, temperatura do globo negro, e velocidade do vento, pressão parcial de vapor, e radiação solar. A análise de variância foi baseada no método dos quadrados mínimos e a comparação de médias foi feita pelo teste de Tukey com nível de 5% de significância. Os resultados mostraram que as cabras do semi-árido brasileiro mantiveram estável a produção de calor metabólico ao longo do dia, no entanto, quando expostas a radiação solar direta, aumentaram consideravelmente a perda de calor latente na superfície cutânea e reduziram a produção de calor metabólico em aproximadamente 24%, numa tentativa de manter a temperatura interna constante

Choque séptico é caracterizado por desequilíbrio entre o transporte e o consumo de oxigênio, podendo acarretar hipóxia tecidual. A disfunção microcirculatória, característica cardinal da fisiopatologia do choque séptico, causa má distribuição de fluxo sanguíneo microvascular e, consequentemente, shunt de oxigênio, disóxia tissular e, teoricamente, diminuição no consumo de oxigênio (VO2) pela célula. No presente estudo, foi investigada a associação entre alterações microcirculatórias causadas pela sepse e o consumo de oxigênio em pacientes pediátricos. Dezessete crianças com choque séptico ressuscitadas foram estudadas em quatro momentos durante a internação na unidade de terapia intensiva (dentro de 24, 48 e 72 horas após a admissão ou diagnóstico de choque e após a resolução deste, antes da extubação traqueal). A microcirculação sublingual foi avaliada utilizando o método de imagem Sidestream dark field (SDF) e o VO2 foi calculado através da calorimetria indireta. Outras variáveis hemodinâmicas, como transporte de oxigênio, índice cardíaco, pressão arterial invasiva, lactato arterial e saturação venosa central, foram coletadas. Embora as variáveis hemodinâmicas tenham se mantido em níveis satisfatórios, graves alterações na microcirculação foram visualizadas, especialmente na densidade de vasos pequenos perfundidos (DVPP), na proporção de vasos pequenos perfundidos (PVPP) e no índice de fluxo microvascular (MFI). Foram encontradas assosciações significativas entre o VO2 e os parâmetros da microcirculação: dVO2 e dDVPP (&#946; coefficient= 6,875; p<0,001), dVO2 e dPVPP (&#946; coefficient=92,246; p<0,001) e dVO2 e dMFI (&#946; coefficient=21,213; p<0,001). Não foram encontradas correlações entre as alterações microcirculatórias e as outras variáveis. Em conclusão, este estudo mostrou que pacientes pediátricos com choque séptico apresentaram grave disfunção microvascular e que o fluxo microcirculatório alterado estava associado ao VO2, podendo estar implicado na fisiopatologia da disóxia tecidual da sepse. / Septic shock is characterized by the imbalance between oxygen delivery and consumption leading to tissue hypoxia. Microcirculatory dysfunction, a key element of septic shock pathogenesis, elicits maldistribution of microvascular blood flow and consenquently oxygen shunt, tissue oxygenation debt and, theoretically, impaired oxygen consumption (VO2). In this study, it was investigated if there is an association between microcirculatory changes and VO2 in pediatric patients with septic shock. Seventeen resuscitated patients with septic shock were studied in four moments (within 24hr, 48hr and 72hr of the admission or diagnosis of shock and after its resolution, prior to extubation). Sublingual microcirculation was evaluated using Sidestream dark field (SDF) imaging and VO2 was measured directly by indirect calorimetry. Other hemodynamic variables, like cardiac index, oxygen delivery, invasive arterial pressure, arterial lactate and central venous oxygen saturation were also recorded. Although global hemodynamic variables were within satisfactory ranges, microvascular variables were markedly altered, especially microvascular flow index (MFI), proportion of perfused small vessels (PPV) and perfused small vessel density (PVD). Significant associations between oxygen consumption and microcirculatory parameters were found: dVO2 and dPVD (&#946; coefficient= 6.875; p<0.001), dVO2 and dPPV (&#946; coefficient=92.246; p<0.001) and dVO2 and dMFI (&#946; coefficient=21.213; p<0.001). There was no correlation between microcirculatory alterations and other variables in this study. In conclusion, this study showed that pediatric patients with septic shock presented severe microcirculatory dysfunction and abnormal microvascular blood flow could be associated to oxygen consumption.

Doctor of Philosophy / Department of Horticulture, Forestry, and Recreation Resources / Candice A. Shoemaker / The objectives of this study were to determine exercise intensity of common gardening tasks in older adults and to investigate if older gardeners meet the physical activity (PA) recommendations (intensity and time) through their daily gardening. Kinds of gardening tasks, body postures, and bodily pain while gardening of older gardeners were investigated and the possibility of gardening as a predictor for a physically active lifestyle and life satisfaction in older adults was determined. Older participants were randomly recruited from the community of Manhattan, KS. To determine the exercise intensity of gardening, the heart rates of older adults were measured by radiotelemetry during gardening or garden tasks, and then oxygen uptake and energy expenditure were measured via indirect calorimetry using a submaximal graded exercise test. Overall health conditions by the Short Form 36 Health Survey (SF-36), hand functions by hydraulic hand dynamometer and pinch gauge, and bone mineral density (BMD) by dual-energy x-ray absorptiometry were measured. An observational study and weekly logs were conducted to study kinds of gardening, postures, and bodily pain of older gardeners. The Community Healthy Activities Model Program for Senior (CHAMPS) questionnaire was used to measure leisure-time PAs (frequency per week of all PAs and calories expended per week in all PAs). In conclusion, the nine gardening tasks were found to be low to moderate intensity PA in healthy older adults (1.6 ± 3.6 METs). Gardening observed was moderate intensity (3.8 ± 1.4 METs) PA in older adults and the subjects met the PA recommendation, which is at least 30 minutes of moderate intensity PA on most days of the week through their daily gardening (moderate intensity; average 33 hrs/wk in May and 15 hrs/wk in June and July). The older gardeners showed higher values for hand function and some SF-36 domains (physical function, bodily pain, and physical summary) than older non-gardeners. Gripping, stooping, lifting, stretching, walking, standing, kneeling, sitting, and squatting were observed while older adults gardened and lower back pain was the main bodily pain reported. Furthermore, gardening was found to be a predictor for leading a physically active lifestyle and high life satisfaction in older adults.

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Consistently, reports in the literature have identified that a sedentary lifestyle contributes to the progression of a range of chronic degenerative diseases. The measurement of energy expenditure and physical activity pattern in children is a challenge for all professionals interested in paediatric health and from a broader perspective, the public health fraternity charged with considering longer term health consequences of physical inactivity. The primary objective of this thesis was to identify a suitable indirect and objective measurement technique for the assessment of energy expenditure and physical activity pattern in children. The ideal characteristics of such a technique are that it should be reproducible and have been validated against a criterion reference method. To achieve this goal, a series of methodological studies were undertaken (Chapters II and III). This work was essential to increase accuracy during the individualised laboratory calibration process and further minimise prediction errors when analysing data from 7 days of monitoring under free-living conditions in the second part of the study (Chapters IV and V). In the first study to verify the combined effect of body position, apparatus and distraction on children's resting metabolic rate (RMR), experiments were carried out on 14 children aged 8-12 (mean age = 10.1 years ± 1.4). Each participant underwent 2 test sessions, one week apart under three different situations: a) using mouthpiece and nose-clip (MN) or facemask (FM); b) sitting (SEAT) or lying (LY) and c) TV viewing (TV) or no TV viewing. In the first session, following 20 min rest and watching TV, the following protocol was used: LY: 20 min - stabilisation; 10 min using MN and 10 min using FM. Body position was then changed to seated: 20 min stabilisation; 10 min using FM; 10 min using MN. In the second session, FM and MN order was changed and participants did not watch TV. Data were analysed according to the eight combinations among the three studied parameters. Repeated measures ANOVA indicated statistically significant differences for &VO2 (p=0.01) and RMR (p=0.02), with TVMNSEAT showing higher values than TVFMLY. Bland-Altman analysis showed a bias for &VO2, &VCO2, RQ and RMR between TVFMLY and TVMNSEAT of -17.8±14.5 ml.min-1, -8.8±14.5 ml. min-1, 0.03±0.05 and -115.2±101.9 kcal.d-1, respectively. There were no differences in RMR measurements due to body position and apparatus when each variable was isolated. Analyses of distraction in three of four combinations indicated no difference between TV and no TV. In summary, different parameter combinations can result in increased bias and variability and thereby reported differences among children's RMR measurement. The second study dealt with treadmill adaptation and determination of self-selected (SS) walking speed. Assessment of individual and group differences in metabolic energy expenditure using oxygen uptake requires that individuals are comfortable with, and can accommodate to, the equipment being utilised. In this study, a detailed proposal for an adaptation protocol based on the SS was developed. Experiments were carried out on 27 children aged 8-12 (mean age = 10.3±1.2 yr). Results from three treadmill tests following the adaptation protocol showed similar results for step length with no significant differences among tests and lower and no statistically significant variability within- and between-days. Additionally, no statistically significant differences between SS determined over-ground and on a treadmill were verified. These results suggest that SS speed determined over-ground is reproducible on a treadmill and the 10 min familiarisation protocol based on this speed provided sufficient exposure to achieve accommodation to the treadmill. The purpose of the third study was to verify within- and between-day repeatability and variability in children's oxygen uptake ( &VO2), gross economy (GE) [ &VO2 divided by speed] and heart rate (HR) during treadmill walking based on SS. 14 children (mean age = 10.2±1.4 yr) undertook 3 testing sessions over 2 days in which four walking speeds, including SS, were tested. Within- and between-day repeatability was assessed using the Bland and Altman method and coefficients of variability (CV) were determined for each child across exercise bouts and averaged to obtain a mean group CV value for &VO2, GE and HR per speed. Repeated measures ANOVA showed no statistically significant differences in within- or between-day CV for &VO2, GE or HR at any speed. Repeatability within and between-day for &VO2, GE and HR for all speeds was verified. These results suggest that submaximal &V O2 during treadmill walking is stable and reproducible at a range of speeds based on children's SS. In the fourth study, the objective was to establish the effect of walking speed on substrate oxidation during a treadmill protocol based on SS. Experiments were carried out on 12 girls aged 8-12 (mean age = 9.9±1.4 yr). Each participant underwent 2 test sessions, one week apart. Workloads on the treadmill included 2 speeds slower than SS (1.6 [V1] and 0.8 km.h-1 [V2] slower than SS), SS (V3), and a speed 0.8 km.h-1 faster than SS (V4). Indirect calorimetry from respired gas measurements enabled total fat (FO) and carbohydrate (CHO) oxidation rates to be calculated according to the non-protein respiratory quotient (Peronnet and Massicote, 1991) and percentage of CHO and FO calculations using equations from McGilvery and Goldstein (1983). Repeated measures ANOVA followed by a Tukey Post Hoc test (p< 0.05) was used to verify differences in CHO and FO rates among speeds. Paired T-test was used to verify differences in CHO and FO rates between tests per velocity. The reliability between-day was assessed using intraclass correlation coefficient (ICC). Results showed significant differences for CHO among all speeds, as well as significant differences for FO between V1 and V2 against V3 and V4 in both tests. Analyses between trials per velocity showed no significant substrate use differences as well as acceptable reliability. At the self-selected speed (V3) there was an accentuation in FO reduction as well as an increase in CHO oxidation. The purpose of the fifth study was to determine whether there were differences in substrate oxidation between girls (G) and women (W) during a treadmill protocol based on SS. Experiments were carried out on 12 G aged 8-12 (mean age = 9.9±1.4 yr) and 12 W aged 25-38 (mean age = 32.3±3.8 yr). The treadmill protocol included 6 min workloads followed by 5 min rest periods. Workloads included 2 speeds slower than SS (1.6 (V1) and 0.8 km.h-1 (V2) slower than SS), SS (V3), and a speed 0.8 km.h-1 faster than SS (V4). Total fat and carbohydrate (CHO) oxidation rates were calculated from indirect calorimetry according to the non-protein respiratory quotient. Repeated measures ANOVA followed by a Tukey Post Hoc test was used to verify intra-test differences in CHO and fat oxidation rates among speeds. Intergroup differences were analysed using paired T-test. Fat utilisation in W achieved a plateau at a relative velocity 0.8 km.h-1 slower than SS, but for G, fat utilisation increased until SS, and then stabilised upon reaching the higher velocity. CHO oxidation curves rose abruptly above V2 for W, while for G the acute increase occurred after SS (V3). Collectively, these results indicate that as walking intensity increases G are able to meet the energy demands of the work by increasing fat oxidation together with the increased CHO oxidation up to SS. In contrast for W, increasing CHO oxidation is associated with an early decrease in fat utilisation at a velocity slower than the self-selected speed. The sixth study dealt with validation of indirect techniques for the measurement of energy expenditure in free-living conditions against the DLW technique. Experiments were carried out on 19 children aged 8-12 (mean age = 10.3±1.0 yr). To indirectly predict energy expenditure 12 different procedures were used. Only one procedure, combining activity and heart rate (AHbranched), was based on a group equation, the others were based on individualised regression. Three of the individually-based techniques were able to accurately predict energy expenditure in free-living conditions. These tecniques were HRPAnetRMR using HRnet [HR exercise minus sleep HR (SHR)] against PAnet (measured PA exercise minus measured RMR) and upper and lower body equations corrected by RMR; HRPAnet4act using the same procedure but corrected by the mean resting &VO2 for 4 resting activities [(4act) = supine watching TV, sitting watching TV, sitting playing computer games and standing], and HRPALBnet4act using only lower body activities and corrected by 4act. HRPAnetRMR was only slightly more accurate than HRPAnet4act and HRPALBnet4act, but this technique is only adjusted by RMR whereas the other two are heavily dependent on more complex laboratory calibration. Bland and Altman (1986) analyses showed no significant differences between AHbranched predicted and measured TEE using the DLW technique. A SEE of 79 kcal.d-1 and a mean difference of 72 kcal.d-1, with a 95% CI ranging from -238 to 93.9 kcal.d-1 was found. In addition, no significant differences between predicted HRPAnetRMR and measured TEE using DLW were found, showing an SEE of 99 kcal.d-1 and a mean difference of -67 kcal.d-1, and a 95% CI ranging from -276.6 to 141.9 kcal.d-1. AHbranched and HRPAnetRMR were both valid and similarly suitable for the prediction of energy expenditure in children under free-living conditions. Significant associations between DLWAEE and the after-school time window indicated that this time window as an important discretionary period representative of children physical activity. However, the duration of the after-school time windows should be more carefully considered. Accelerometer data showed a better association between the largest after-school time window (3.5 hr) and measured TEE. The final study, completed with 19 children aged 8-12 (10.3±1.0 yr) highlighted, under laboratory conditions across a range of walking and running speeds, the inadequacy of the use of the standard MET in children. This traditional approach overestimates energy expenditure with an increased difference linearly related to speed increments. Minute-by-minute analyses of 7 days of free-living monitoring showed an average overestimation of 64 minutes per day for moderate-to-vigorousphysical- activity (MVPA) using the standard MET compared with the individually measured MET. For all intensities, these differences were statistically significant (p< 0.001). The second part of this study showed a variability of 20% in the average time spent at MVPA when comparing HR I 140 bpm and HR > 50%P &VO2 (P &VO2 = the highest &VO2 observed during an exercise test to exhaustion). Results of the current study compared to observations in the literature showed that HR I 140 bpm consistently estimates lower MVPA time than HR > 50%P &VO2. When these two PA indices were compared with individual and standard MET measured minute-byminute, statistically significant differences were verified among all of them at MPA, but no differences were verified at VPA, except between individual and standard METs. However, whether each one of the PA indices used are under- or overestimating time at MVPA is still debatable due to the lack of a gold standard. Finally, each index used in this study classified different numbers of participants as achieving the PA target of 60 min.d-1. The wide variability between indices when attempting to classify children who are achieving the recommended target is cause for great concern because habitually these indices are utilised as screening tools in paediatric and public health settings and used to guide behavioural interventions.