Submitted by Erika Demachki (erikademachki@gmail.com) on 2014-12-29T15:35:49Z No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertação - Letícia de Almeida Leandro - 2013.pdf: 2111763 bytes, checksum: c992b02afb4a59e318265d06577e6454 (MD5) / Approved for entry into archive by Erika Demachki (erikademachki@gmail.com) on 2014-12-29T15:40:58Z (GMT) No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertação - Letícia de Almeida Leandro - 2013.pdf: 2111763 bytes, checksum: c992b02afb4a59e318265d06577e6454 (MD5) / Made available in DSpace on 2014-12-29T15:40:58Z (GMT). No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertação - Letícia de Almeida Leandro - 2013.pdf: 2111763 bytes, checksum: c992b02afb4a59e318265d06577e6454 (MD5) Previous issue date: 2013-02-28 / The analysis of the energetic and respiratory metabolism of Taenia crassiceps cysticerci innoculated into the encefalum of female BALB/c mice was performed. After 30 days of infection the mice were treated with low dosages (3.0 and 6.0 mg/kg) of albendazole and praziquantel aiming the observation of the alterations caused by a hostile environment on the parasite's metabolism. The aim of this study was to detect the influence of low dosages of anti-helminthic drugs on the production of organic acids related to the intermediary metabolism (oxaloacetate, malate, fumarate, succinate, citrate and α-ketoglutarate), carbohydrates metabolism (pyruvate, lactate, propionate, oxaloacetate and malate) and fatty acids and proteins catabolism (β-hydroxibutyrate, acetoacetate, acetate, α-ketoglutarate and oxalate) by T. crassiceps cysticerci innoculated into the CNS of BALB/c mice. Regarding the glycolisis, it was possible to detect lactate in all samples which shows that this parasite performs a lactic fermentation. As to the intermediary metabolism it was possible to detect oxaloacetate, citrate, malate, succinate, fumarate and α-ketoglutarate which indicates that the cysticerci may have used the tricarboxilic acid cycle (TCA) to produce energy. However facing the results found the evidences show that the cysticerci used a metabolic pathway denominated reversion of the TCA for energy production which presents as main end products propionate and acetate. There were no significant differences between the groups treated with the anti-helminthic and the control one except that the group treated with 6.0 mg/kg of albendazole presented acetate as main end product of the TCA reversion while the other groups presented propionate. In the groups treated with praziquantel, both concentrations, there was no production of acetate and in the 6.0 mg/kg of praziquntel there was no production of β-hydroxibutyrate wheter in the groups treated with both concentrations of albendazol there was an increase in the production of this organic acid which indicates the maximization of the energetic pathway of fatty acids oxidation and/or of the proteic catabolism. It was possible to conclude that T. crassiceps cysticerci when exposed to the conditions of this study preferred an anaerobic energy production pathway and probably performed the catabolism of carbohydrates, proteins and fatty acids. / Realizou-se a análise do metabolismo energético e respiratório de cisticercos de Taenia crassiceps inoculados no encéfalo de camundongos BALB/c fêmeas. Após 30 dias de infecção, esses camundongos foram tratados com baixas doses (3,0 e 6,0 mg/kg) de albendazol e praziquantel visando observação de alterações provocadas no metabolismo do parasito. O objetivo deste trabalho foi detectar a influência de baixas doses desses anti-helmínticos na produção dos ácidos orgânicos do metabolismo intermediário (citrato, α-cetoglutarato, succinato, fumarato, malato, oxaloacetato), de carboidratos (piruvato, lactato, oxaloacetato, malato, acetato e propionato), de ácidos graxos e proteínas (β-hidroxibutirato, acetoacetato, acetato, α-cetoglutarato e oxalato) realizados por cisticercos de T. crassiceps implantados no SNC de camundongos BALB/c fêmeas. Com relação a glicólise, detectou-se lactato em todas as amostras mostrando que esse parasito fez fermentação lática; em relação ao metabolismo intermediário foi possível detectar os ácidos orgânicos citrato, α-cetoglutarato, succinato, fumarato, malato e oxaloacetato, indicando que os cisticercos apresentaram elementos do ciclo do ácido cítrico para produzir energia. Porém diante dos resultados encontrados a principal evidência é de que esses cisticercos utilizaram uma via denominada reversão do ciclo do ácido cítrico para produzirem energia, obtendo como produtos finais propionato e acetato. Não se observou diferenças significativas entre os grupos que sofreram tratamentos e os grupos controle exceto pelo fato de que, o grupo tratado com albendazol 6,0 mg/kg de peso do camundongo teve como principal produto final da reversão do ciclo do ácido cítrico, o acetato, ao contrário dos demais grupos que tiveram o propionato; nos grupos tratados com praziquantel não houveram a produção de acetato como nos demais grupos e no grupo tratado com praziquantel 6,0 mg/kg não houve a produção de β-hidroxibutirato; nos grupos tratados com albendazol observou-se um aumento na produção de β-hidroxibutirato indicando que esse grupo maximizou a via de produção energética através da β-oxidação de ácidos graxos e/ou através do catabolismo proteico.Concluímos que os cisticercos de T. crassiceps, nas condições do presente trabalho, deram preferência para uma via de produção energética anaeróbia e provavelmente catabolizaram, além de carboidratos, proteínas e ácidos graxos para produzirem ATP.

O pacu, Piaractus mesopotamicus, é um caracídeo neotropical autóctone, de hábito alimentar onívoro, largamente utilizado na aqüicultura brasileira. No entanto, alguns aspectos da sua nutrição permanecem controversos ou desconhecidos. O presente trabalho visou determinar a exigência em proteína, energia e aminoácidos para juvenis de pacu por meio da avaliação de parâmetros de desempenho, composição corporal e hematológicos em dois ensaios distintos. No primeiro ensaio, juvenis de pacu (15.5±0.4 g) foram alimentados durante 10 semanas, duas vezes ao dia, com dietas contendo teores de proteína bruta (PB) entre 22 e 38% (incremento de 4%) e energia digestível (ED) entre 2600 e 3400 kcal kg-1 (incremento 200 kcal kg-1), em um delineamento totalmente aleatorizado, em esquema fatorial 5 × 5 (n=3). O ganho de peso (GP) e a taxa de crescimento específico (TCE) aumentaram significativamente com a elevação da PB dietética. A retenção de nitrogênio (RN) e a taxa de eficiência protéica (TEP) diminuíram (p<0,05) com o aumento da proteína das dietas em todos os níveis de ED testados. As concentrações de ED afetaram (p<0,05) a concentração de umidade, proteína, gordura, matéria mineral e energia bruta corporal e o fator de condição. A taxa de eficiência econômica foi influenciada pela proteína (p<0,05) e energia digestível (p<0,05) das dietas e pela interação entre ambas (p<0,05). Entretanto, o índice de lucratividade econômica somente foi influenciado pela concentração de proteína bruta (p<0,05) das dietas. Efeito significativo (p<0,05) dos tratamentos nos parâmetros hematológicos foi registrado para taxa de hemoglobina, concentração de hemoglobina corpuscular média, proteína plasmática total e glicose plasmática. A exigência mínima para ganho de peso de juvenis de pacu é 27% de PB, com relação PB:ED ótima de 92,9 mg PB kcal-1 ED. As concentrações protéicas e energéticas estudadas não causaram prejuízos a saúde dos peixes. A exigência em lisina dietética para juvenis do pacu Piaractus mesopotamicus, com peso inicial de 4,3 g, foi determinada fornecendo cinco dietas experimentais isonitrogenadas (32% proteína bruta) contendo caseína, gelatina e L-aminoácidos cristalinos com níveis crescentes de lisina (0,90; 1,17; 1,44; 1,69 e 1,96% da matéria seca da dieta) por 74 dias. Cada dieta foi aleatoriamente designada para grupos (n=4) de 18 peixes alimentados três vezes ao dia até a aparente saciedade. Não foram observados mortalidade ou sinais de deficiência além da redução no desempenho dos peixes alimentados com dietas deficientes ou com excesso de lisina. O peso final (PF), GP, TCE, índice de eficiência alimentar (IEA), TEP, VPP, composição corporal, morfometria e hematologia foram afetados (p<0,05) pelas concentrações de lisina dietética. A análise polinomial quadrática indicou a exigência em lisina em 1,45, 1,51 e 1,43% da matéria seca da dieta, respectivamente para GP, IEA e VPP. Foram obtidos valores de lipídios e proteína corporais significativamente mais altos nos peixes alimentados com as dietas com 0,9 e 1,96% de lisina. Foi então determinado que a exigência em lisina para juvenis de pacu é de 1,5% da matéria seca da dieta. / Pacu, Piaractus mesopotamicus, a neotropical, omnivorous native characin, widely used in Brazilian aquaculture. However, some aspects of your nutrition are controversial or ignored. This work aimed at determining dietary protein, energy and amino acids requirements of juvenile pacu through the evaluation of performance, body composition and hematological parameters. In a first trial, pacu juveniles (15.5±0.4 g) were fed twice a day for 10 weeks until apparent saciety with diets containing crude protein (CP) from 22 to 38 % (intervals of 4%) and digestible energy levels among 2600 and 3400 kcal kg-1 (intervals of 200 kcal), in a totally randomized experimental design, 5 × 5 factorial scheme (n=3). Weight gain (WG) and specific growth rate (SGR) increased significantly with amount dietary CP. Nitrogen retention (NR) and protein efficiency ratio (PER) decreased (p<0.05) with increasing dietary protein at all tested levels of dietary energy. DE levels affected (p<0.05) whole body moisture, protein, fat, ash, gross energy and condition factor. Economic efficiency ratio was influenced by dietary protein (p<0.05) and digestible energy (p<0.05), and by interaction between the two factors. However, income was affected by dietary protein level (p<0.05) alone. Significant effect of treatments in hematological parameters were registered for mean corpuscular hemoglobin concentration, total plasma protein and plasma glucose. Crude protein requirements and optimum protein:energy ratio for weight gain of juveniles of pacu were 271 g kg-1 and 92.9 mg kcal-1, respectively. All dietary crude protein and digestible energy levels studied did not pose harms to fish health. Dietary lysine requirement of juvenile pacu Piaractus mesopotamicus (4.3 g) was determined by feeding five isonitrogenous (32% crude protein) amino acid test diets containing casein, gelatin and Lcrystalline amino acids with graded levels of lysine (0.90, 1.17, 1.44, 1.69 and 1.96% of dry diet) for 74 days, three times a day until apparent satiation, to groups (n=4) of 18 fish in a totally randomized design trial. No mortality or nutritional deficiency signs were observed; reduced growth was recorded for fish fed diets with either the lower or the higher lysine contents. Final weight (FW), WG, SGR, feed efficiency (FE), PER, NR, proximate whole body composition, morphometry and hematology were affected (p<0.05) by dietary lysine concentrations. Seconddegree polynomial regression analysis of WG and PER data indicated dietary lysine requirement of 1.45, 1,51 and 1.43% of dry diet, respectively to WG, FE and PPV. Significantly higher lipid and protein contents values were obtained for whole body of fish fed the diets with 0.9 and 1.96% of lysine. Lysine requirement of juvenile pacu was determined as being 1.5% of dry diet.

Cette thèse présente les adaptations spécifiques des hibernants ‘food-storing’ qui s’alimentent au cours de l’hibernation, et les conséquences de la qualité du régime alimentaire sur leur cycle annuel. Tandis que les espèces ‘fat-storing’ jeûnent pendant toute l’hibernation, les ‘food-storing’ alternent jeûnes courts et réalimentations. L’adiponectine stimulerait la lipolyse pendant l’hibernation contribuant ainsi à la cétogenèse. Le maintien d’un système digestif fonctionnel conduisant à la sécrétion d’incrétines, permet l’absorption optimale de nutriments lors des courtes euthermies inter-torpeurs. Une absorption accrue de glucose en particulier permettrait de restaurer la glycémie et les réserves de glycogène. Par ailleurs, un régime appauvri en protéines et enrichi en lipides induit un engraissement augmenté en période pré-hibernatoire provoquant une moindre utilisation de la torpeur donc une perte de masse accrue lors de l’hibernation, et une baisse du succès reproducteur. / This thesis presents the specific adaptations of food-storing hibernators that feed during hibernation, and the impact of diet quality on their annual cycle. In contrast to the fat-Storing species which fast during hibernation, the food-storing presents metabolic responses to an alternation of short fasting phases and hyperphagia. These responses involve one hand use of fat reserves during hibernation contributing to ketogenesis, which would be induced by adiponectin. On the other hand, maintaining a functional digestive system leading to the secretion of incretins, permits optimal nutrient absorption in the short inter-torpor euthermia. Increased glucose uptake in particular would restore body reserves to spare. Moreover, a lean protein diet enriched in fat and induces increased in body mass in pre-hibernation period causing reduced use of torpor thus an increased loss of mass during hibernation, and decreased reproductive success.

L’anorexie mentale (AN), un trouble du comportement alimentaire multifactoriel, se traduit par une perte de poids. La sévère dénutrition retrouvée dans l’AN est associée à des altérations métaboliques induisant une dérégulation de l’axe intestin cerveau. Les mécanismes physiopathologiques sont encore mal connus. Le travail de cette thèse était de mieux appréhender les dysfonctions de l’axe intestin cerveau en évaluant le métabolisme protéique de divers tissus (hypothalamus, côlon et estomac) dans un modèle murin d’anorexie par une approche protéomique. Le premier travail a permis de mieux caractériser le modèle d’anorexie nommé activity-based anorexia (ABA) en fonction du sexe. Puis les différentes analyses protéomiques ont permis de constater une adaptation tissu dépendant des mécanismes régulant l’équilibre énergétique, avec une activité cérébrale potentiellement augmentée au détriment des fonctions digestives. Chez les souris femelles ABA, il a été constaté une augmentation d’expression de protéines mitochondriales au niveau de l’hypothalamus et à l’inverse, une diminution du métabolisme protéino-énergétique au niveau colique avec un rôle de la voie de signalisation mTOR. L’autophagie était augmentée dans ces deux tissus. Ensuite, nous avons démontré un ralentissement de la vidange gastrique secondaire à la dénutrition, et l’analyse protéomique a permis de constater une augmentation du stress oxydant au niveau de l’antre des souris ABA femelles. Ces altérations peuvent contribuer aux troubles fonctionnels gastro intestinaux. En conclusion, nos études soulignent des mécanismes d’adaptation tissu dépendants dans l’anorexie, qui devront être ultérieurement approfondis. / Anorexia nervosa, a multifactorial eating disorder, is a major public health problem and results in a severe body weight loss. The severe malnutrition observed in anorectic patients is associated with metabolic alterations inducing disturbance of the gut-brain axis. However, involved mechanisms remained poorly understood. The aim of the present thesis was to better understand the alterations of the gut-brain axis in the activity-based anorexia (ABA) model by evaluating the protein metabolism of various tissues (hypothalamus, colon and stomach) by proteomic approach. Firstly, we have better characterized the response to ABA model according to sex. Then, different proteomic analyses were performed using female C57BL/6 mice. Our results revealed a tissue-dependent adaptation of protein and energy metabolism with an increased hypothalamic activity and a decrease in the gastrointestinal tract. Indeed, ABA mice exhibited an increased expression of proteins involved in mitochondrial metabolism at the level of the hypothalamus, and conversely a decrease of proteins involved in protein and energy metabolism in colonic mucosa with a key role of the mTOR signaling pathway. Both in hypothalamus and colon, autophagy was increased. We were also able to show that gastric emptying was delayed in ABA mice that is mainly due to malnutrition. In addition, proteomic analysis revealed an increase in gastric oxidative stress in female ABA mice. These alterations may contribute to the gastrointestinal functional disorders frequently described in anorexia nervosa. In conclusions, our study underlined tissue-dependent adaptive metabolic process during anorexia that should be further explored.

AMP-activated protein kinase (AMPK) is an important metabolic sensor in eukaryotic organisms and it plays an important role in regulating energy homeostasis, at both the cells and the whole organism. AMPK controls glucose and lipid metabolism by direct stimulation of enzymes or by long term stimulation of the gene expression of energy metabolism. Skeletal muscles significantly contribute to the total body weight and metabolic rate and to the maintenance of glucose homeostasis. Due to the ability of the muscle to increase energy expenditure to 95% of whole-body energy expenditure, could be the proper development and programming of metabolism in the early postnatal period crucial for the further development of the organism in adulthood. Early postnatal development leads to substantial changes in energy requirements of the body and this suggests the significant involvement of AMPK in this period. The aim of this thesis was to study the activity and expression of isoforms of the catalytic subunit of AMPK in skeletal muscle during early postnatal development of both mouse strains A/J and C57BL/6 that differ in the development of diet-induced obesity. The next task was to analyze the expression of selected genes involved in energy metabolism - GLUT4, PGC-1α and UCP3 that AMPK regulates. It was found that the...

Le fer est un micronutriment important pour la croissance et le développement des plantes. Il agit comme cofacteur pour plusieurs enzymes et il est important pour des processus tels que la photosynthèse et la respiration. Souvent, le Fe dans le sol n’est pas bio-disponible pour la plante. Les plantes ont développé des stratégies pour solubiliser le Fe du sol pour le rendre disponible et assimilable pour elles. Il y a deux stratégies, la première est caractéristique des dicotylédones et la seconde est caractéristique des monocotylédones. Le modèle utilisé dans cette étude est une culture cellulaire de Solanum tuberosum. Une partie de la recherche effectuée a permis la mesure d’activité et d’expression relative de certaines enzymes impliquées dans le métabolisme énergétique et la fourniture de précurseurs pour la synthèse d’ADN : la Nucléoside diphosphate kinase, la Ribonucléotide reductase, la Glucose 6-phosphate déshydrogénase et la 6-Phosphogluconate déshydrogénase dans les cellules en présence ou en absence de Fe. Chez certains organismes, la déficience en Fe est associée à une perte de croissance qui est souvent liée à une diminution de la synthèse d’ADN. Chez les cultures de cellules de S. tuberosum, les résultats indiquent que la différence de biomasse observée entre les traitements n’est pas due à une variation de l’activité ou l’expression relative d’une de ces enzymes. En effet, aucune variation significative n’a été détectée entre les traitements (+/- Fe) pour l’activité ni l’expression relative de ces enzymes. Une autre partie de la recherche a permis d’évaluer l’activité des voies métaboliques impliquées dans la stratégie 1 utilisée par S. tuberosum. Cette stratégie consomme des métabolites énergétiques: de l’ATP pour solubiliser le Fe et du pouvoir réducteur (NAD(P)H), pour réduire le Fe3+ en Fe2+. Des études de flux métaboliques ont été faites afin d’étudier les remaniements du métabolisme carboné en déficience en Fe chez S. tuberosum. Ces études ont démontré une baisse du régime dans les différentes voies du métabolisme énergétique dans les cellules déficientes en Fe, notamment dans le flux glycolytique et le flux de C à travers la phosphoenolpyruvate carboxylase. En déficience de Fe il y aurait donc une dépression du métabolisme chez S. tuberosum qui permettrait à la cellule de ralentir son métabolisme pour maintenir sa vitalité. En plus des flux, les niveaux de pyridines nucléotides ont été mesurés puisque ceux-ci servent à réduire le Fe dans la stratégie 1. Les résultats démontrent des niveaux élevés des formes réduites de ces métabolites en déficience de Fe. L’ensemble des résultats obtenus indiquent qu’en déficience de Fe, il y a une baisse du métabolisme permettant à la cellule de s’adapter et survivre au stress. / Iron is an important micronutrient for plant growth and development. It participates as a cofactor for several enzymes and is important for processes such as photosynthesis and respiration. Often soil Fe is not bioavailable to the plant. Plants have developed strategies to solubilize the Fe in the soil to make it available and easy to assimilate. There are two strategies, the first is characteristic of dicotyledones and the second is characteristic of monocotyledones. The model used in these studies is a cell culture of Solanum tubersoum. A first part of the research involved the study of expression and activity of enzymes required in energy metabolism and the provision of precursors for DNA synthesis: Nucleoside dehydrogenase, Ribonucleotide reductase, Glucose 6-phohate dehydrogenase and 6-Phosphogluconate dehydrogenase. In several organisms, Fe deficiency induces a loss of biomass which is often associated with a decrease in DNA synthesis. In S. tuberosum cell cultures, the results indicate that the loss of biomass observed in Fe deficiency is not linked to a change in the activity or relative expression of these enzymes. Indeed, no significant changes were detected between treatments (+/- Fe) for activity or relative expression. In another part of the research, we evaluated the activity of the metabolism pathways involved in strategy 1, which is used by S. tuberosum. This strategy consumes energetic metabolites: ATP to solubilize Fe and reducing power (NAD(P)H) to reduce the Fe3+ to Fe2+. Metabolic flux studies were done to investigate the alterations of carbon metabolism during Fe deficiency in S. tuberosum. These studies demonstrated that in Fe deficient cells, there is a decrease in the fluxes of some pathways of energy metabolism. Particularly, in the glycolytic flux and the anaplerotic flux of PEPC. Under Fe deficiency there would be a depression of metabolism in S. tuberosum which would allow the cell to slow its metabolism to maintain its vitality. In addition to the fluxes, the levels of pyridine nucleotides were measured since they serve to reduce Fe in the strategy 1. The results show an increase in the reduced forms of these metabolites during Fe deficiency. All results together point out that during Fe deficiency the metabolism decreases, allowing the cell to survive and adapt to the stress.

Les cœurs univentriculaires sont des cardiopathies congénitales, non compatibles avec la vie. Fontan a transformé le pronostic de ces patients en restaurant une circulation pulmonaire passive. Malheureusement, le pronostic à long terme reste péjoratif. Pour diminuer la morbi-mortalité de ces patients, nous avons développé un protocole chirurgical de pré-conditionnement, permettant une totalisation de Fontan percutanée adaptative aux résistances pulmonaires et aux capacités de travail du ventricule. Nous avons pu observer que les chaînes respiratoires mitochondriales n’ont pas la possibilité d’adapter leur production énergétique face au travail. Le déséquilibre de la balance Nitroso-Redox engendrera le mitohormésis et l’hormésis, puis inéluctablement surviendra un remodelage myocardique mal adaptatif. La seule possibilité pour améliorer le pronostic à long terme de ces patients sera d’augmenter les capacités oxydatives myocardiques du ventricule unique ou de créer une pompe pulmonaire. / Univentricular hearts are congenital heart diseases, not compatible with life. Fontan transformed the prognosis of these patients by restoring a passive pulmonary circulation. Unfortunately, the long-term prognosis remains pejorative. To reduce the morbidity and mortality of these patients, we developed a surgical preconditioning protocol, allowing a percutaneous Fontan totalization, adaptive to pulmonary resistance and to the working capacity of the ventricle. We observed that the mitochondrial respiratory chains do not have the ability to adapt their energy production toward work. The desequilibrium of the Nitroso-redox balance will generate mitohormesis and hormesis, then will inevitably occur a poorly adaptive myocardial remodeling. The only way to improve the long-term prognosis of these patients is to increase the myocardial oxidative capacities of the single ventricle or to create a pulmonary pump.

Made available in DSpace on 2014-07-29T15:26:21Z (GMT). No. of bitstreams: 1 Tatiane Luiza da Costa.pdf: 1394288 bytes, checksum: cde5d3e8ab1f315e5e9ac6dd8d167bd0 (MD5) Previous issue date: 2011-09-16 / Leishmaniasis is a disease of the endothelial reticulum system and is caused by protozoans form the Leishmania genus. The promastigote forms of Leishmania sp degradate carbohydrates through the glycolityc pathway and its first reaction occurs inside the glicosome. As they use as energy source glucose, aminoacids and lipids from the culture media the excreted/secreted (E/S) products may be useful as indicative of the energetic and metabolis profile of the parasite and of its biological disturbances. The biochemical analysis of Leishmania metabolism is important to upgrade the parasitological studies with techniques which are adequate to the evaluation of the biochemical profile of the host-parasite relationship towards a better understanding of how these organisms are able to survive in such diverse environments such as the digestory tract of the insect or inside vertebrate hosts tissues. The aim of this study was to perform a study of the energetic metabolism of species and isolates of Leishmania spp. Therefore the in vitro analysis of E/S metabolic products from the parasite within the culture media during logarithmic and stationary growth phases was performed through chromatographic (HPLC) methods. The spectrophometry was used to glucose measurement and the nuclear magnetic resonance (NMR) was implanted aiming the study of the parasite&#8223;s metabolism. Through HPLC it was possible to verify a significant difference in the E/S of organic acids from the carbohydrates metabolism (pyruvate and lactate), intermediary metabolism (citrate, &#945;-ketoglutarate, succinate, fumarate, malate and oxaloacetate) and from the fatty acids oxidation (&#946;-hydroxibutyrate) between differents species and isolates of Leishmania spp. Through the enzymatic spectrophotometric analysis we verified the different concentrations of glucose. The NMR through 13C and 1H detected the following metabolites in pellet and supernatant: alanine, lactate, acetate, oxalate, pyruvate, succinate, glicerol, CO2, ureia and aminoacids (glutamine, valine and arginine). Therefore, through the several methods it was possible to observe that there is difference of E/S metabolites between the isolates and strains of Leishmania spp that utilize several biochemical pathways to energy production and the identification of these pathways may be used to identify each isolate and also establish metabolic differences in the host-parasite relationship. / A leishmaniose é uma doença do sistema mononuclear fagocitáio causada por protozoários do gênero Leishmania. As formas promastigotas de Leishmania sp degradam carboidratos pela via glicolítica e a primeira reação ocorre dentro do glicossomo. Utilizando como fonte de energia a glicose, aminoácidos e lipídeos presentes no meio de cultura, os produtos excretados/secretados (E/S) podem ser indicadores do perfil metabólico energético parasitário e de distúrbios biológicos. A análise bioquímica do metabolismo de Leishmania vem atualizar os estudos parasitológicos com técnicas adequadas à avaliação do perfil bioquímico na relação parasito-hospedeiro, levando ao melhor entendimento da maneira como esses organismos conseguem sobreviver em ambientes diversos como o trato digestório do hospedeiro invertebrado ou nos tecidos do hospedeiro vertebrado. O objetivo deste trabalho foi o estudo do metabolismo energético de diferentes espécies de Leishmania. Para tanto, foi realizada a análise in vitro de metabólitos E/S pelo parasito em meio de cultura durante as fases logarítmicas e estacionárias do crescimento, e quantificados por cromatografia líquida de alta eficiência (CLAE). A espectrofotometria foi utilizada para a dosagem de glicose. O método de ressonância magnética nuclear foi implantado para o estudo do metabolismo parasitário. Pela CLAE verificou-se diferença nos produtos E/S dos ácidos orgânicos do metabolismo de carboidratos (piruvato e lactato), do metabolismo intermediário (citrato, &#945;-cetoglutarato, succinato, fumarato, malato e oxaloacetato) e do metabolismo dos ácidos graxos (&#946;-hidroxibutirato) entre diferentes espécies e isolados de Leishmania. Pela análise espectrofotométrica enzimática verificou-se diferença na concentração de glicose na cepa M2903 de L.(V.) braziliensis. Pela RMN de 13C e 1H detectaram-se os seguintes metabólitos no sedimento e sobrenante: alanina, lactato, acetato, oxalato, piruvato, succinato, glicerol, CO2, uréia e aminoácidos (glutamina, valina e arginina). Entretanto, através de vários métodos, foi possível observar que há diferenças na E/S de metabólitos entre diferentes cepas e isolados de Leishmania spp que utilizam diversas vias bioquímicas para a obtenção de energia e a identificação dessas vias poderá ser utilizada para investigar a atuação de fármacos anti-parasitários. Mudanças bioquímicas no metabolismo podem indicar possíveis adaptações do parasito às diferentes condições ambientais e fisiológicas durante o ciclo de vida e essas características bioquímicas individuais poderão ser utilizadas na identificação de cada isolado, além de estabelecer diferenças metabólicas na relação parasito-hospedeiro.

L’abeille domestique (Apismellifera) a un rôle majeur dans les écosystèmes naturels et agronomiques mais est exposée à un nombre croissant de pressions environnementales (nouveaux parasites, xénobiotiques, variations climatiques et malnutrition). Dans ce contexte, la compréhension des phénomènes impliqués dans les réponses au stress ainsi que leurs coûts associés devient cruciale pour mieux appréhender l’impact de ces pressions sur les abeilles. L’émergence d’un stress perturbe généralement l’homéostasie de l’organisme qui doit mettre en place une cascade d’adaptations physiologiques et comportementales pour le surmonter. Cependant, du fait de son mode de vie social, il est raisonnable de penser que les réponses vont se faire dans l’intérêt du groupe et non plus seulement dans l’intérêt de l’individu. Afin de caractériser les réponses au stress et de déterminer leur spécificité en fonction de la nature du stimulus (xénobiotiques, immunitaire, thermique, social), j’ai adopté une approche multidisciplinaire en ciblant l’identification des modifications i) physiologiques associées à la division du travail, ii) du métabolisme énergétique, et iii) comportementales. J’ai démontré quequelque soit leur rôle social (nourrice, gardienne, butineuse), les abeilles répondent de la même manière à un stress donné, si celui-­ci est écologiquement pertinent (hyperthermie et stress immunitaire mais pas xénobiotique). Une tendance à la diminution des ressources énergétiques a également été observée suite à un stress suggérant une modification des performances comportementales. Afin de vérifier cela, je me suis concentrée sur l’activité de butinage; le vol chez les insectes étant un des processus physiologiques les plus coûteux du règne animal. Une altération des performances de butinage a été mise en évidence chez les abeilles soumises à un stress immunitaire avec une réorientation des préférences de butinage au dépens du pollen, plus coûteux àc ollecter et moins riche en ressource énergétique que le nectar ; ceci probablement pour pallier au coût énergétique du stress. En revanche, en réponse àune hyperthermie, une augmentation de l’activité de butinage a été observée mais sans engendrer un coût supplémentaire au niveau des ressources collectées.Ces résultats sont discutés à la lumière du coût énergétique du stress et des conséquences potentielles sur les performances des abeilles, qui infine pourrait perturber l’homéostasie énergétique de la colonie. / Honeybees (Apis mellifera), which play an important role in natural and agronomic ecosystems, are exposed to a growing number of environmental pressures(new parasites, pesticides, climatechangeand poor nutrition). In this context, deciphering the mechanisms underlying stress responses and their costs becomes crucial to better understand theim pact of these pressures. Stress usually represents a challenge to the homeostasis of a norganism. In response, a cascade of physiological and behavioural adaptations enables the organism to cope with the stress. However, dueto their sociallife style, we could suggest that stress response in honeybees will occurin the interest of the colony and not only in the interest of the individual. To characterise the stress response and determine its specificity according to the stimulus (xenobiotic, immune, thermal, social), I developed a multidisciplinary approach to identify changes in i) task-­related physiology, ii) energetic metabolism, and iii) behaviour. I demonstrated that, regardless of their social function (nurse, guard, forager), bees respond in the sameway to a given stress, if itis ecologically-­relevant (heat and immune stress but not pesticides). Atendencytoward decreas ingenergetic resources was also observed following stress exposure, which suggests changes in behavioural performance.In order to test this hypothesis, I analysed changes in foraging activity in response to stress, as insect flight is one of the most costly physiological processes in the animal kingdom. I found that for aging performances were affected by animmune stress : bees changed their foraging preferences at the expense of pollen, probably to reduce the stress energetic cost, given that pollen is more costly to collect and provides alower energetic return than nectar. In contrast, in response to heat stress, an increase in colony for aging activity was observed, without an additional cost on resource collection. These results are discussed in the light of stress energetic cost and its potential consequences onhoneybee performances, which could disrupt the colony’s energetic homeostasis.

Le ratio métabolique (MR) est un marqueur du métabolisme cérébral. Dans notre travail, nous avons démontré sa valeur pronostique chez 68 patients victimes d’une hémorragie sous-arachnoïdienne anévrysmale grave. En effet, une diminution du MR sous le seuil de 3,35 traduit un phénomène d’hyperglycolyse relative, dont le nombre d’événement est prédictive d’un pronostic défavorable avec une excellente sensibilité et spécificité. L’obtention de ces résultats est rendue possible, notamment après une phase de validation dans un modèle animal de procédures permettant de limiter les effets de facteurs pré-analytiques critiques. Ces résultats permettent d’envisager une étude pour savoir si l’intégration de ce marqueur dans la stratégie de prise en charge du patient, permet de modifier son devenir fonctionnel. Après avoir validé analytiquement les mesures de pyruvate, glucose et lactate impliquant la technique de microdialyse, nous avons étudié sur une cohorte de patients graves aSAH, modeste (n=18 patients) s’il existait des phénomènes d’hyperglycolyse et leur corrélation avec le pronostic. Dans notre série, à la différence de l’approche globale (cathétérisme de la veine jugulaire), un phénomène d’hyperglycolyse conduirait vers un bon pronostic. En fait, l’approche par microdialyse donne une information sur le métabolisme énergétique localisé à l’implantation de la sonde, alors que le MR donne une information globale, ce qui est probablement le facteur le plus important expliquant la différence d’interprétation entre les 2 approches. En l’absence d’outils de traitement de données et d’algorithmes de décision clinique validés, la microdialyse ne donne pas à l’heure actuelle, une valeur individuelle diagnostique ou pronostique. Un des résultats très prometteurs de ce travail, est la mise en évidence d’un phénomène d’hyperglycolyse relative globale lors du vasospasme, rapidement réversible chez les patients ayant bien évolué, alors qu’il perdure de nombreuses heures après le vasospasme chez les patients ayant évolué de manière péjorative. Ces résultats nécessitent d’être reproduits sur un nombre plus significatifs de patients, ce qui permettrait une confirmation radiologique du vasospasme de manière plus précoce afin de confirmer son importance, sa localisation et l’éventualité de le traiter rapidement / The metabolic ratio (MR) is an index of the brain energetic metabolism. In our study, we have demonstrated its prognostic value for 68 poor grade patients aneurysmal subarachnoid hemorrhage (aSAH): a MR below the threshold value of 3.35 reflects a phenomenon of global cerebral hyperglycolysis which, if repeated, is predictive of a bad outcome. These results were made possible after validation step in an animal mode which allowed to control the critical pre-analytical factors. Our results pave the way for a clinical study aiming to determine if taking into account the MR will help to improve the functional outcome of the aSAH patients. In another approach, based on the use of cerebral microdialysis, we have studied, in an 18 patients cohort, and after an analytical validation of a new biochemical analysis, if such cerebral hyperglycolysis phenomenon was a encountered in this cohort, if these was a correlation with the patients’ outcome. In contrast with the previous 68 aSAH patients, this hyperglycolysis phenomenon appears linked to a good outcome. This apparent discrepancy may be due the difference in the anatomical giving a more localized information on the brain metabolism than the jugular approach used for the MR determination. The most interesting of our results is the correlation found between hyperglycolysis and cerebral vasospasm. If conformed with a larger cohort of aSAH patients, the use of MR could allow an earlier detection and treatment of cerebral vasospasm