La première partie de ce travail est réalisé dans le but de préparer des produits pouvant jouer un rôle très important de l’imagerie cellulaire des lipopolysaccharides sur la surface des cellules bactériennes, tout en utilisant des méthodes fiables telle que la ‘’chimie click’’. Dans un premier temps, nous avons synthétisé une première génération d’outils fluorescents à base de rhodamine B et fluorescéine modifiée par une fonction oxyde de nitrile. Dans un deuxième temps nous avons cherché les meilleures conditions d’applications de la chimie click 3+2 sans cuivre, entre la fonction oxyde de nitrile et le motif saccharidique complémentaire portant une fonction vinylique. Finalement, nous avons appliquée, avec succès, la méthode click avec cuivre sur plusieurs types de bactéries portant sur leur membrane cellulaire des lipopolysaccharides ayant un motif ‘’Kdo’’ fonctionnalisé par un groupement azoture déjà synthétiser au sein de notre équipe, et une sonde fluorescente porteuse d’une fonction alcyne. Une nouvelle génération d’outils de marquage saccharidique est en cours de finalisation dans le but d’optimiser les conditions finales, tel que le dérivé de ‘’Kdo’’ fonctionnalisé cette fois-ci par des dérivées cycliques, bicycliques et aromatiques porteurs d’une fonction alcyne ou alcène, pour réaliser les derniers essais de marquage in vitro. Le stress oxydant est lié au vieillissement des cellules et à de nombreuses maladies: cardiovasculaire, cancer, diabète, Alzheimer… Il est dû à un déséquilibre entre le système oxydant / antioxydant au niveau des cellules, et se caractérise par la présence principalement de substance radicalaires réactives oxygénées. Dans le but d'identifier le taux de substances réactives oxygénées dans les cellules, le travail dans la deuxième partie de la thèse reposait sur la synthèse multi étape d'une molécule fluorescente dérivée de la coumarine. Le composé ciblé est le peroxyde d'oxygène, connu sous le nom d'eau oxygénée. Possédant un ester vinyl-boronique, notre molécule sera oxydée et réarrangée en aldéhyde conduisant à la condensation intramoléculaire avec le groupement aminé adjacent pour former un cycle indolique, libérant ainsi de la fluorescence. La recherche des conditions optimales de la dernière étape de la voie synthétique sont toujours en cours d’optimisation. Dans le futur, on cherchera les conditions optimales de la dernière étape de la synthèse de la sonde spécifique au peroxyde d’oxygène. Cette molécule est d'une importance remarquable comme sonde du stress oxydant. En réussissant cette étape, on pourra avoir en main une bibliothèque de ‘’KDO’’ fonctionnalisé afin d’avoir des résultats satisfaisants in vivo. / The first part of this work is done in order to prepare products that play a very important role in cellular imaging lipopolisaccharides on the surface of bacterial cells, while using reliable methods such as '' click chemistry ''. Initially, we synthesized the first generation of tools based fluorescent rhodamine B and fluorescein -modified nitrile oxide function. In a second step we sought the best possible applications of click chemistry 3+2 copper free, between the nitrile oxide function and the additional saccharide unit with a vinyl function. Finally, we applied successfully, the click method with copper on several types of bacteria on their cell membrane lipopolysaccharides having a pattern Kdo functionalized azide group already synthesized within our team, and a probe carrying a fluorescent alkyne. A new generation of tools saccharide marking is being finalized in order to optimize the final terms, such as derivative functionalized Kdo this time by cyclic, bicyclic and aromatic derivatives holders of an alkene or alkyne function to perform final testing of in vitro labeling. Oxidative stress is linked to cell aging and many diseases: cardiovascular disease, cancer, diabetes, Alzheimer's ... It is due to an imbalance between oxidant system/antioxidant in cells, and is characterized mainly by the presence of radical substance reactive oxygen. In order to identify the rate of reactive oxygen substances in cells, work in the second part of the thesis based on multi- step synthesis of a fluorescent molecule derived from coumarin. The target compound is oxygen peroxide, known as the name of hydrogen peroxide. Having a vinyl boronic ester, this molecule will be oxidized and rearranged aldehyde leading to intramolecular condensation with the adjacent amino group to form an indolique ring , thereby releasing fluorescence. The search for optimal conditions for the last step of the synthetic pathway is still being optimized. In the future, the optimal conditions for the last step of the synthesis of specific probe oxygen peroxide are sought. This molecule is remarkable importance as a probe of oxidative stress. By passing this step, we will have on hand a library of KDO functionalized to have satisfactory results in vivo.

Lorsque l’anesthésie par inhalation ne peut être utilisée chez le rat, la combinaison de kétamine et de xylazine est l’alternative la plus fréquemment utilisée. Les doses administrées peuvent varier selon le protocole expérimental. En présence de fièvre, d’infections ou de processus tumoral accompagné de fièvre, la pharmacocinétique de ces drogues peut être modifiée. Ce projet porte sur l’évaluation des changements physiologiques, hématologiques, biochimiques et pharmacocinétiques chez le rat Sprague Dawley lors d’anesthésie avec le mélange kétamine-xylazine suite à l’administration de trois doses différentes de lipopolysaccharide (LPS). Après l’administration de LPS, une anesthésie à la kétamine-xylazine fut induite chez des rats Sprague Dawley. Des prélèvements sanguins périodiques ainsi que des mesures des paramètres physiologiques furent effectués afin d’évaluer l’effet du LPS sur la pharmacocinétique des deux drogues ainsi que sur les paramètres biochimiques et hématologiques. Les différentes doses de LPS ont causé certaines modifications notamment en produisant une baisse marquée de la saturation en oxygène et de l’albumine sérique, une augmentation de la durée d’anesthésie ainsi que des lésions hépatiques mineures. Les paramètres pharmacocinétiques de la kétamine furent peu altérés par l’administration de LPS tandis qu’une diminution de la clairance et une augmentation de l’aire sous la courbe (AUC) furent observées pour la xylazine dans les groupes ayant reçu les doses moyenne et élevée de LPS. Ces résultats montrent que les doses de xylazine doivent être adaptées en présence de LPS pour permettre une anesthésie de courte durée et des changements physiologiques et biochimiques moindres lorsqu’elle est administrée avec de la kétamine. / When inhalation anesthesia cannot be used in laboratory rats, ketamine-xylazine combination is the most frequent alternate regimen. The administrated doses can vary according to the experimental protocol. During fever episodes, infections or tumoral process, the pharmacokinetics of these drugs can be modified. This project focuses on the evaluation of the physiological, hematological, biochemical and pharmacokinetics changes in Sprague Dawley rats during ketamine-xylazine anesthesia, after administration of three different doses of lipopolysaccharide (LPS). After administration of LPS to Sprague Dawley rats, ketamine-xylazine anesthesia was induced. Periodic blood samplings and monitoring of physiologic parameters were made in order to evaluate the effect of LPS on ketamine-xylazine pharmacokinetics and hematological and biochemical parameters. The different LPS doses caused specific parameter modifications including a marked decrease of oxygen blood saturation and serum albumin, a longer anesthesia duration and minor hepatic lesions. No significant modifications of pharmacokinetics parameters of ketamine were observed. An increase of area under curve (AUC) and a decrease of xylazine clearance were noted in groups who received medium and large doses of LPS. These results show that that xylazine doses need to be adapted in the presence of LPS, to allow a shorter duration anaesthesia and lesser physiological and biochemical changes when administered with ketamine.

Thèse numérisée par la Division de la gestion de documents et des archives de l'Université de Montréal

Les syndromes septiques restent à ce jour un problème majeur de santé publique. Une importante immunodépression est observée lors des syndromes septiques, affectant notamment les lymphocytes T, acteurs majeurs de la réponse immunitaire. En effet, après avoir subi une apoptose massive ils présentent d'importantes altérations fonctionnelles et phénotypiques, associées à un mauvais pronostic. Cependant les mécanismes impliqués dans le développement de ces altérations lymphocytaires induites par le sepsis (ALIS) sont encore mal connus. Le but de ce travail était d'étudier ces mécanismes à travers la mise au point de modèles ex-vivo. Nous avons optimisé un test de mesure de la réponse proliférative des lymphocytes T, utilisable pour le diagnostic des immunodéficiences primaires, comme pour l'étude expérimentale et clinique des ALIS. Nous avons développé un premier modèle ex vivo, reposant sur l'incubation de cellules mononuclées du sang périphérique (PBMC) en présence de LPS, suivie d'une stimulation spécifique des lymphocytes T via leur TCR. Ce modèle récapitule des mécanismes indirects potentiellement impliqués dans l'induction des ALIS, impliquant les monocytes. Puis, nous avons mis au point un modèle qui repose sur l'incubation de PBMC en présence d.IL-10, suivie d'une stimulation des lymphocytes T par des anticorps anti-CD2/CD3/CD28. Ce modèle pourrait reproduire des mécanismes directs et indirects impliqués dans l'induction des ALIS. Nos résultats nous ont permis d'améliorer la compréhension des mécanismes en jeux dans les ALIS et d'en souligner la complexité. Les modèles ex-vivo présentés pourraient permettre d'évaluer de nouvelles stratégies thérapeutiques / Sepsis remains a major public healthcare issue. During sepsis, an important immunodepression develops, affecting particularly the T lymphocytes, major players of the immune response. Following a massive apoptosis, T lymphocytes display important functional and phenotypical alterations, which are associated with higher risk of secondary infections and higher mortality. Mechanisms involved in the induction of such alterations are not fully understood. The aim of this study was to analyze those mechanisms through the development of ex vivo models. We optimized a test to measure T lymphocytes proliferative response, which can be used for the diagnosis of primary immunodeficiencies, as well as to clinically and experimentally study sepsis induced T lymphocytes alterations (SILA). First, we set up a model consisting in an incubation of PBMC (peripheral blood mononuclear cells) with LPS followed by specific T lymphocytes stimulation via its TCR. This model recapitulates indirect mechanisms likely to participate in SILA induction, mediated by monocytes. Then, we set up a model consisting in PBMC incubation with IL-10 followed by T lymphocytes stimulation with anti-CD2/3/28 antibody coated beads. This model may recapitulate direct and indirect mechanisms involved in SILA induction. Our results allowed us to improve the understanding of the mechanisms involved in SILA induction and to highlight its complexity. The ex-vivo models that we developed could be used for the evaluation of new therapeutic strategies

Acute Respiratory Distress Syndrome (ARDS) following hemorrhagic shock/resuscitation (S/R) is an important contributor to late morbidity and mortality in trauma patients. S/R promotes ARDS by inducing oxidative stress that primes cells of the innate immune system for excessive responsiveness to small inflammatory stimuli, termed the “twohit” hypothesis. Activated alveolar macrophages (AM) play a central role and when recovered from S/R animals exhibit an exaggerated responsiveness to lipopolysaccharide (LPS) with increased activation of the proinflammatory transcription factor NF-κB, and augmented expression of cytokines. LPS triggers AM signalling through Toll like receptor 4 (TLR4), which resides in plasma membrane lipid rafts. The objective of this work is to define cellular mechanisms of macrophage priming by oxidative stress following shock resuscitation. The main hypothesis investigated is that altered cellular distribution of TLR4 can lead to macrophage priming and antioxidant resuscitation strategies can diminish these effects. AM of rodents, exposed in vivo to oxidant stress following S/R, increase their surface levels of TLR4, which in turn results in augmented NF-κB translocation in response to small doses of LPS. Furthermore, in vitro H2O2 treatment of RAW 264.7 macrophages results in similar TLR4 surface translocation. Depletion of intracellular calcium, disruption of the cytoskeleton or inhibition of the Src kinases prevents the H2O2-induced TLR4 translocation, suggesting the involvement of receptor exocytosis. Further, fluorescent resonance energy iii transfer between TLR4 and lipid rafts as well as biochemical raft analysis demonstrated that oxidative stress redistributes TLR4 to surface lipid rafts. Preventing the oxidant-induced movement of TLR4 to lipid rafts using methyl-ß-cyclodextrin precluded the increased responsiveness of cells to LPS after H2O2 treatment. Further, AM priming by oxidative stress can be diminished by early exposure to resuscitation regimens with direct or indirect systemic antioxidant effects, such as 25% albumin, N-acetylcysteine and hypertonic saline. Hyperosmolarity was found to modulate AM TLR4 gene and protein expression. Collectively, these studies suggest a novel mechanism whereby oxidative stress might prime the responsiveness of cells of the innate immune system. Targeting the TLR4 signalling pathway early during shock resuscitation may represent an anti-inflammatory strategy able to ameliorate late morbidity and mortality following S/R.

Vibrio vulnificus are marine bacteria causing fatal septicemia through wound infections or consumption of contaminated seafood. V. vulnificus is an excellent model for the study of surface polysaccharides, as it is capable of synthesizing capsular polysaccharide (CPS), lipopolysaccharide (LPS) and exopolysaccharide (EPS). V. vulnificus strains exhibit a multitude of carbotypes that evolve through unknown mechanisms. CPS is a confirmed virulence factor, but the genetics of its biosynthesis are unknown. The main objective of these experiments was to gain insight into the biosynthesis, regulation and evolution of ATCC 27562 outer surface polysaccharides. A miniTn10 transposon (Tn) system was used for mutagenesis and single insertions were confirmed through Southern analysis. A novel 25 kb CPS biosynthesis locus was identified through sequencing of regions surrounding Tn insertions; a region encoding putative LPS core biosynthetic functions was identified adjacent to the CPS cluster. The CPS locus contained features of O-antigen biosynthetic loci and was unusual in carrying characteristics of both group I and IV capsular biosynthetic loci. Mutations in this region resulted in elimination of CPS and LPS, and both were shown to be dependent on the activity of the polymerase Wzy. Evidence is presented here supporting horizontal transfer (HT) as a contributor to V. vulnificus CPS evolution. CPS regions of V. vulnificus 27562, YJ016 and CMCP6 contain strain specific genes surrounded by conserved regions, suggestive of HT. Moreover, a CPS locus virtually identical to that of 27562 was discovered in Shewanella putrefaciens strain 200. 27562 CPS is distinctive as it contains N-acetylmuramic acid. Genes encoding murA and murB activities were identified within the cluster and shown to be functionally redundant, supporting HT acquisition of this region. A screen of V. vulnificus gDNA library using CPS biosynthesis and transport mutants identified a cyclic diguanylate cyclase, dcpA. dcpA-mediated increase in cyclic diguanylate lead to EPS production, rugosity phenotypes and enhanced biofilm formation. Interestingly, virulence and motility were not affected suggesting complexity of cyclic diguanylate regulation in V. vulnificus, supported by the large number of cyclic diguanylate related proteins in Vulnificus strains.

As vacinas pertussis celulares apresentam certa reatogenicidade e as acelulares, menos reatogênicas, têm seu uso restrito, devido a seu alto custo. O Instituto Butantan desenvolveu uma vacina pertussis celular (Plow), com baixo teor de lipopolissacarídeo e outra acelular (Pa), por metodologia simples e econômica. Essas preparações, combinadas aos toxóides diftérico e tetânico (DTPlow e DTPa, respectivamente), foram comparadas à DTP tradicional, com diferentes adjuvantes: vitamina A, surfactante pulmonar, BCG, monofosforil lipídeo A (MPL) e Al(OH)3. A resposta humoral em camundongos foi semelhante para as diferentes formulações e independente do adjuvante utilizado. As vacinas induziram níveis equilibrados de IgG1/IgG2a anti-pertussis e a DTPlow mostrou-se menos reatogênica, induzindo níveis significativamente menores de IL-6 sérica. A adição de MPL sugeriu tendência de proteção contra a colonização nasotraqueal no grupo imunizado com DTPa e levou à indução de IFN-g nos grupos imunizados com DTP e DTPa, sugerindo possível atividade imunomodulatória para Th1. / The whole cell pertussis vaccines present some reactogenicity and the acellular, less reactogenic, have prohibitive use due to its high cost. Instituto Butantan developed a less reactogenic whole cell pertussis vaccine (Plow), with low lipopolysaccharide content and an acellular vaccine (Pa), by simple and economic methodology. These preparations, combined to diphtheria and tetanus toxoids (DTPlow and DTPa, respectively), were compared with the traditional DTP, with different adjuvants: vitamin A, pulmonary surfactant, BCG, monophosphoryl lipid A (MPL) and Al(OH)3. The humoral immune response induced in mice by the different vaccine formulations, was similar and independent of the adjuvant used. The vaccines induced balanced levels of IgG1/IgG2a anti-pertussis and DTPlow showed to be less rectogenic, inducing lower levels of serum IL-6. The use of MPL suggested to induce higher protection against nasotracheal colonization in DTPa group and induced IFN-g in the DTP and DTPa groups, suggesting a possible immunemodulatory activity for Th1.

Este trabalho foi desenvolvido com o objetivo de verificar a possível influência do estresse gestacional, pela indução de Lipopolissacarídeo (E. coli) sobre a relação materno filial e desempenho do cordeiro, assim como o estresse da desmama sobre o desempenho em confinamento e nas características da carcaça. A execução do experimento aconteceu de dezembro de 2013 a julho de 2014 no anexo do Laboratório de Biometeorologia e Etologia localizado no setor de caprino e ovinocultura da Prefeitura do Campus Administrativo da Universidade de São Paulo em Pirassununga-SP. Foram utilizadas 43 ovelhas multíparas, mestiças da raça Santa Inês e 55 cordeiros, crias provenientes da IATF dessas ovelhas com sêmen de macho White Dorper. O delineamento experimental utilizado foi o inteiramente casualizado (DIC) tendo um fatorial 3 x 2 como o esquema de distribuição dos tratamentos composto por 3 grupos durante a gestação (controle (Gc), aplicação de LPS aos 70 (±7 dias) dias de gestação (Gi) e o aos 120 (±7 dias) dias de gestação (Gf)) combinando com 2 tipos de desmama (desmama controlada (Dc) e desmama abrupta (Da)). As médias foram comparadas pelo teste F e teste t de Student (PDIFF), com um nível de significância de 5%. No primeiro experimento o objetivo foi analisar a influência do estresse, gerado pelo uso de LPS, em diferentes períodos gestacionais sobre os níveis de cortisol e temperatura retal das ovelhas, o comportamento materno filial, os níveis de cortisol logo após o parto, peso ao nascer (PN) dos cordeiros e o tempo gasto para a primeira mamada. Os animais foram delineados n os 3 grupos: Gc, Gi e Gf. Na fase intermediária da gestação, analisando o ritmo circadiano do cortisol e da TR para os tratamentos Gc e Gi pela interação entre tratamento e tempo foram observados maiores valores de cortisol e TR para Gi (P<0,05). Na fase final com a mesma interação entre tratamento e tempo, os maiores valores de cortisol e temperatura durante o ritmo circadiano foram de Gf (P<0,05). Para as avaliação do comportamento materno filial, as observações das posturas e das atividades tanto das ovelhas quando dos cordeiros, foram analisadas pela interação entre tratamento e tipo de parto (P<0,05). Também houve interação entre tratamento e o tipo de parto para o tempo a primeira mamada tendo o tratamento Gf em situação de parto duplo apresentado o maior tempo, não sendo diferente apenas do tratamento Gi também com parto duplo (P<0,05). Analisando o PN, a interação aconteceu entre tratamento e sexo com os menores valores observados para o tratamento Gf quando comparados aos demais tratamentos (P<0,05). O LPS alterou os níveis de cortisol durante o ritmo circadiano, assim como o estresse durante a gestação influenciou no comportamento materno filial. Os animais submetidos ao estresse durante o terço final da gestação apresentaram maior gasto de tempo para ingerir colostro e pesos mais leves no nascimento. No segundo experimento o objetivo foi avaliar a eficiência da desmama controlada ou abrupta em reduzir o impacto do estresse durante a gestação em relação ao desempenho de cordeiros. Foram utilizados 47 cordeiros, 22 machos e 25 fêmeas distribuídos em 6 tratamentos: GcDc, GcDa, GiDc, GiDa, GfDc e GfDa. Foram analisados os níveis de cortisol sanguíneo e a TR da desmama controlada e na comparação entre os dois tipos de desmama, peso a desmama (PD) e os dados de desempenho em confinamento pela ingestão de matéria seca (IMS), conversão alimentar (CA), eficiência alimentar (EA), ganho médio diário (GMD) e medidas pós-abate. Houve efeito fixo do estresse durante a gestação e dos dias para os níveis de cortisol durante a DC (P<0,05), para a temperatura retal foi observado efeito fixo de sexo, dia e tempo (P<0,05). No dia de entrada dos animais no confinamento houve efeito fixo de tempo (colheitas durante o dia) e tipo de desmama, com maiores valores de cortisol para a primeira colheita do dia (P<0,05) e para a desmama abrupta quando comparada com a controlada (P<0,05), neste mesmo dia para a TR foi observado efeito fixo de tempo, sendo a primeira colheita menor que as demais (P<0.05). Nas colheitas de 24 horas e 48 após a entrada dos animais no confinamento Da apresentou maior nível de cortisol do que DC (P<0,05), e feito contrario foi observado para TR. Dc apresentou maior peso a desmama que Da (P<0,05). Houve interação entre estresse na gestação e sexo para IMS e GMD, com menores valores para machos de Gf (P<0,05), não havendo diferenças significativas para CA e EA em nenhum tratamento. Nas análises pós - abate, os menores valores foram observados para Gf e Da. A desmama controlada pode reduzir os efeitos do estresse durante a gestação, melhorando o desempenho de cordeiros, assim como cordeiros machos quando desafiados na fase final da gestação são mais suscetíveis que fêmeas. / This study was developed with the objective to investigate the possible influence of gestational stress by inducing Lipopolysaccharide (E. coli) on maternal and filial relationship performance of the lamb, as well as the stress of weaning on feedlot performance and on characteristicsin housing. Running the experiment took place from December 2013 to July 2014 in the Annex to Biometeorology and Ethology Laboratory located in goat and sheep production sector of Administrative Campus Hall, Universidade de São Paulo in Pirassununga-SP. We used 43 multiparous sheep, crossbred Santa Inês and 55 lambs, calves from AIFT those sheep with male White Dorper semen. The experimental design was completely randomized having a 3 x 2 factorial as the distribution pattern of treatments consists of 3 groups during pregnancy (control (Cg), application of LPS to 70 (± 7) days of gestation (Ig) and at 120 (± 7) days of gestation (Fg)) combined with two types of weaning (controlled weaning (Cw) and abrupt weaning (Aw)). Means were compared by F test and Student\'s t test (PDIFF), with a 5% significance level. In the first experiment the objective was to analyze the influence of stress, generated by the use of LPS in different gestational periods on the levels of cortisol and body temperature of sheep, filial maternal behavior, cortisol levels immediately after parturition, birth weight (BW) of the lambs and the time spent for the first feeding. The animals were allotted in three groups: Cg, Ig and Fg. In the middle stage of pregnancy, analyzing the circadian rhythm of cortisol and RT for the treatment Cg and Ig by the interaction between treatment and time showed higher cortisol levels to Ig and RT (P<0.05). In the final stage with the same interaction between treatment and time increased cortisol and temperature values during the circadian rhythm were Fg (P<0.05). For the evaluation of filial maternal behavior, observations of the postures and activities of both the sheep when the lambs were analyzed by the interaction between treatment and type of delivery (P<0.05). There was also an interaction between treatment and the type of parturition to the time the first feeding with the Fg treatment in dual labor situation presented as long, not being different only Ig treatment also double parturition (P<0.05). Analyzing the BW, the interaction occurred between treatment and sex with the lowest values observed for Fg treatment compared to the other treatments (P<0.05). LPS altered levels of cortisol during the circadian rhythm, and stress during pregnancy the influenced filial maternal behavior. The animals subjected to stress during the final third of gestation had higher expenditure of time to ingest colostrum and lighter weights at birth. In the second experiment the objective was to evaluate the efficiency of the subsidiary or abrupt weaning to reduce the impact of stress during pregnancy in relation to the performance of lambs. We used 47 lambs, 22 male and 25 female, comprised of 6 treatments: CgCw, CgAw, IgCw, IgAw, FgCw and FgAw. Blood cortisol levels were analyzed and the TR controlled weaning and the comparison between the two types of weaning, weaning weight (WW) and performance data in confinement for dry matter intake (DMI), feed conversion (FC), feed efficiency (FE), average daily gain (ADG) and post-slaughter measures. There was fixed effect of stress during pregnancy and days to cortisol levels during the Cw (P<0.05) for the rectal temperature was observed fixed effect sex, day and time (P<0.05). On receipt of the animals was fixed effect confined in time (during harvest day) and type of weaning, with higher cortisol values for the first harvest day (P<0.05) and the abrupt weaning compared with controlled (P<0.05) in the same day for the TR was observed fixed effect of time, the first harvest less than the others (P<0.05). At harvest 24 hours and 48 hours after entering of animals in the feedlot had higher cortisol levels than Cw (P<0.05) and made opposite was observed for TR. Dc showed higher weaning weight that Aw (P<0.05). There was interaction between stress during pregnancy and sex for DMI and ADG, with lower values for males of Gf (P<0.05), with no significant differences for FC and FE in any treatment. In analysis post - slaughter, the lowest values were observed for Fg and Aw. From Controlled weaning can reduce the effects of stress during pregnancy, improving the performance of lambs males, and lambs when challenged in late pregnancy are more susceptible than females.

Lesão pulmonar aguda é caracterizada por infiltrado de neutrófilos no pulmão, edema, lesão alveolar difusa e alteração da reatividade das vias aéreas. Evidências clínicas e experimentais sugerem que os hormônios sexuais femininos (HSF) modulam a inflamação. Nesse estudo camundongos fêmeas C57BL/6 foram ovariectomizados (OVx) ou falsamente operados, com ovários mantidos (Sham). Após 7 dias foi realizada instilação nasal de LPS (ou salina como controle). Após 4 ou 24 h os experimentos foram conduzidos. Os resultados obtidos 24 h após o LPS mostram aumento de neutrófilos, IL-1b e de óxido nítrico (NO) e redução de IL-10 no pulmão dos animais OVx em relação aos Sham. Também foi observada redução da reatividade traqueal e da resistência e elastância pulmonar dos animais OVx. O tratamento dos animais OVx com 17b-estradiol ou progesterona antes do LPS reverteram esses efeitos. Os dados mostram que 4 h após o LPS a OVx não altera a inflamação e a reatividade das vias aéreas. Pode-se concluir nesse modelo experimental que os HSF exerçam papel de proteção ao organismo. / Acute lung injury is characterized by lung neutrophil infiltration, edema, diffuse alveolar damage and changes in airway reactivity. Clinical and experimental evidences suggest that the female sex hormones (FSH) modulate the inflammation. In this study, female mice C57BL/6 were ovariectomized (OVx) or not (Sham). After 7 days, LPS or saline (as control) were intranasally instillated. After 4 or 24 h the experiments were carried. We observed, 24 h after LPS, increase in neutrophil, IL-1b and nitric oxide (NO) and reduction of IL-10 in the lungs of OVx compared to Sham. We also noted reduced tracheal reactivity and lung resistance and elastance in OVx animals. Treatment of OVx with 17b-estradiol or progesterone before LPS reversed these effects. The data show that 4 h after LPS, the OVx does not promote inflammation and airway reactivity. We can conclude that in our experimental model, FSH exert protective role.

O comportamento maternal (CM) em mamíferos tem características específicas. O período logo após o parto é particularmente sensível a alterações fisiológicas que podem modular a expressão deste comportamento importante. Mudanças comportamentais observadas em animais doentes são consideradas comportamento doentio (CD). A exposição ao LPS, uma endotoxina derivada da parede de uma bactéria gran negativa, durante a gravidez pode causar doenças mentais. A fim de investigar, uma possível relação entre CM e CD, os animais foram tratados com LPS. Para o estudo do CM e agressivo, quarenta ratas foram divididos em quatro grupos, dois controles e dois grupos experimentais, com dez animais cada. O grupo experimental recebeu 100µg/kg de LPS por via i.p, e grupo controle o veículo de endotoxina, após quarenta e oito horas de administração de LPS, ou seja, no quinto dia de lactação, as observações começaram. Para escolha deste dia, ratas virgens e ratas lactantes foram divididas em quatro grupos, dois controles e dois experimentais, com dez fêmeas cada. O peso corporal, consumo de água, ração, e a temperatura corporal foram medidas para cento e vinte horas. As fêmeas do grupo controle foram observadas da mesma forma, mas foram tratados com o veículo do LPS. Observamos que: 1) Em ratas virgens e lactantes o tratamento com LPS modificou a temperatura e peso corporal, consumo de água e ração; 2) No período de lactação houve redução da latência para busca do primeiro filhote. Na prole verificou-se que: 3) Houve alteração no padrão de vocalização dos filhotes cujas mães foram expostas ao LPS no terceiro dia de lactação; 4) houve alteração no burst e fagocitose de enutrofilos no vigésimo primeiro dia de lactação após desafio com a endotoxina indicativo de maior resposta ao LPS. Concluiu-se que a exposição de ratos ao LPS facilita o comportamento maternal, mas promove alterações na sua prole relacionadas à interação entre mãe-filhote e aumento na resposta a um desafio imunológico. / Maternal behavior (MB) in mammals has specific characteristics. The time period just after parturition is particularly sensitive to physiological changes that can modulate the expression of this important behavior. Behavioral changes observed in sick animals, are considered as sick behavior (SB). Exposure to LPS, an endotoxin derived from the wall of a gran negative bacteria, during pregnancy might cause mental diseases. In order to investigate, a possible relationship between MB and SB, animals were treated with LPS. For the study of MB and maternal aggressive behavior, 40 rats were divided in 4 groups, 2 control and 2 experimental groups. The experimental group received 100µg/kg LPS by ip, and control group the vehicle of endotoxin, after 48 hours of LPS administration the observations of SB began. For choice these days, 20 virgin and 20 lactating rats were divided in 4 groups, 2 control and 2 experimental. They received ip 100µg/kg. Body weight, water and feed consumption, and body temperature were measured for 120h. Control females were observed in the same way, but they were treated with vehicle of LPS. The results showed that: 1) In 48 hours after the LPS treatment, virgin and lactating rats showed increased body temperature, loss of body weight, increased water consumption and decreased food consumption, 2) In 48 hours after the treatment with LPS, lactating rats showed reduced latency to retrieve the first pup to the nest. In the offspring of mothers treated with LPS it was found that: 3) Pups form mothers treated with LPS on the 5th day of lactation showed changes in the vocalization pattern; 4) Those pups showed changes in oxidative burst and phagocytosis on 21th day of lactation. It is concluded that exposure of rats to LPS promoted changes in the in the interaction between mother and pups.