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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
61

Réponses microévolutives et coûts adaptatifs de populations de Caenorhabditis elegans exposées à des stress environnementaux / Microevolutionary responses and adaptive costs of Caenorhabditis elegans populations exposed to environmental stress

Dutilleul, Morgan 12 March 2013 (has links)
L'évolution contemporaine des organismes vivants est de plus en plus dépendante des perturbations d'origine anthropique. En particulier, la pollution amplifie l'intensité des pressions de sélection auxquelles sont soumises les populations. Or ces changements peuvent avoir des effets négatifs sur la vie des individus, la démographie des populations, mais aussi, au cours des générations, sur leurs caractéristiques phénotypiques et génétiques. Ainsi en réponse aux pressions de sélection, des changements microévolutifs sont susceptibles de se manifester. Mais ces phénomènes entraînent en parallèle la mise en place de coûts adaptatifs qui fragilise le maintien des populations. Il est donc indispensable de développer nos connaissances sur l'évolution des populations en milieu pollué. Dans ce contexte, cette étude vise à déterminer les réponses microévolutives de populations de C. elegans, exposées à diverses pollutions et à mesurer les coûts adaptatifs de ces microévolutions. Ces populations ont été exposées expérimentalement, durant 22 générations, à une forte concentration en uranium et/ou en chlorure de sodium. Nous avons mis en évidence une différentiation génétique des populations exposées, associée à augmentation de la résistance, au cours du temps. La vitesse des réponses évolutives était dépendante des conditions d'exposition et de leurs effets sur l'expression de la structure génétique des traits d'histoire de vie. Ces phénomènes ont pu être reliés à des coûts adaptatifs, comme une réduction de la fertilité, dans de nouveaux environnements stressants (ex : augmentation de la température) ou en l'absence de stress. Ce projet nous a permis de mieux identifier comment une exposition à un ou deux polluants peut affecter la réponse évolutive de populations de C. elegans et d'évaluer les conséquences sur leur sensibilité. / The contemporary evolution of organisms is largely dependent on anthropogenic disturbances. In particular, the pollution amplifies the intensity of selection pressures suffered by populations. However, these changes may have negative effects on the life of individuals, the demographics of the populations, and its phenotypic and genetic characteristics over generations. Thus, microevolutionary changes are likely to occur in response to selection pressures. These phenomenon lead to collateral damages: adaptive costs. Populations can be more susceptible to many environmental changes. Hence, it is essential to expand our knowledge on the evolution of populations in polluted environment. In this context, our study aims to determine the microevolutionary response of C. elegans populations exposed to different pollutions, and to measure their costs of adaptation. Populations were experimentally exposed for 22 generations to a high concentration of uranium and/or sodium chloride. We confirmed the genetic differentiation between populations with an increase of resistance in populations exposed to different pollutions. The speed of evolutionary responses depended on exposure conditions and their effects on the expression of the genetic structure of life history traits. Microevolutionary changes were linked to costs of adaptation, such as reduced fertility, in stressful novel environments (e.g. fast temperature raise) or in the absence of stress. This project allowed us to better identify how exposure to one or two pollutants affects the evolutionary response of C. elegans populations and evaluate the impact on their sensitivity to environmental conditions.
62

Facilitation and competition in coastal dunes: meta-analysis of determinants of plant interactions / Facilitação e competição em planícies costeiras: meta-análise de fatores determinantes de interações entre plantas

Souza, Luanne Caires da Cruz 23 July 2018 (has links)
Plants established next to other plants may have their performance positively or negatively affected by their neighbors, which characterizes interactions of facilitation and competiton, respectively. Facilitation and competition, however, may occur simultaneously and understanding which one predominates in pairwise interactions under different ecological contexts is important to comprehend the structure of plant communities. According to the stress-gradient hypothesis (SGH), facilitation is expected to prevail in more severe environments, but the outcome of interactions may change depending on features of interacting individuals, such as life form and ontogenetic stages. As harsh conditions and high diversity of plant life-forms are characteristic of coastal dunes, the amount of studies about plant interactions has been rapidly increasing in these ecosystems, with apparent divergent conclusions. However, until now, there is not a systematic and quantitative synthesis about the factors affecting the net outcome of facilitation and competition in these ecosystems. We conducted a meta-analysis to investigate the effects of environmental stress and the simultaneous influence of plant life form and ontogenetic stage on the outcome of facilitation and competition in coastal dunes around the world. We used four performance measures to estimate the outcome of interactions: abundance, survival, growth, and reproduction. Contrary to what was expected by SGH, we found that negative impacts of neighbors on plant reproduction increase towards more arid conditions, but this effect was not observed for other performance measures. Our results also indicate that woody neighbors facilitate the survival of woody seedlings and the reproduction of herbs, while herbaceous neighbors facilitate the growth of other herbaceous plants. Overall, the outcome of plant interactions in coastal dunes depends on the performance variable measured and on both environmental conditions and plant features, indicating an interaction between these factors. Such interaction and different mechanisms underlying facilitation and competition should be more investigated in the future. The global scale of our meta-analysis supports generalization of important processes of succession and conservation in coastal dunes. Benefits of woody neighbors to the survival of woody seedlings corroborate the concept of successional feedbacks in the beach-inland physiognomic gradient, and our results reinforce the use of nurse plants in coastal dunes as a valuable tool to restoration of these endangered ecoystems / Plantas estabelecidas nas proximidades de outras plantas podem ter sua performance afetada positiva ou negativamente por seus vizinhos, caracterizando, respectivamente, interações de facilitação e competição. Considerando que ambas as interações podem ocorrer de forma simultânea, compreender o predomínio de cada uma delas em diferentes contextos ecológicos é fundamental para o entendimento da estrutura de comunidades vegetais. De acordo com a hipótese do gradiente de estresse (HGE), a facilitação tende a predominar em ambientes mais severos, mas o balanço das interações depende ainda das características dos indivíduos envolvidos, como forma de vida e estágio ontogenético. Como condições ambientais severas e alta diversidade de formas de vida vegetais são características de planícies costeiras, o número de estudos investigando interações entre plantas tem aumentado rapidamente nesses ambientes, com conclusões aparentemente divergentes. No entanto, ainda não há uma síntese sistemática e quantitativa dos fatores que afetam o balanço entre facilitação e competição nesses ecossistemas. Nós realizamos uma meta-análise em escala global para investigar os efeitos do estresse ambiental e a influência simultânea da forma de vida e do estágio ontogenético das plantas sobre o balanço entre facilitação e competição em planícies costeiras. Utilizamos quatro variáveis de performance para estimar o balanço de interações: abundância, sobrevivência, crescimento e reprodução. Ao contrário do predito pela HGE, encontramos que impactos negativos de plantas vizinhas sobre a reprodução de outras plantas aumentam com a aridez do ambiente, mas que esse efeito não é observado para outras variáveis de performance. Nossos resultados também mostram que vizinhos lenhosos facilitam a sobrevivência de plântulas lenhosas e a reprodução de plantas herbáceas, enquanto vizinhos herbáceos facilitam o crescimento de outras herbáceas. De modo geral, o balanço das interações depende da variável de performance medida e tanto de condições ambientais quanto de características das plantas, indicando que esses fatores interagem. Tal interação e os diferentes mecanismos subjacentes à facilitação e à competição devem ser melhor investigados no futuro. A maior sobrevivência de plântulas lenhosas na presença de vizinhos lenhosos corrobora a ideia de retroalimentação positiva no processo sucessional que caracteriza o gradiente fisionômico da praia ao interior. Nossos resultados também reforçam o potencial do uso de plantas-berçários como ferramenta para restauração de planícies costeiras degradadas
63

Analyse de la méthylation de l'ADN par séquençage haut-débit chez la Poule / Analyse of the DNA methylation through high-throughput sequencing in Chicken

Mersch, Marjorie 30 October 2018 (has links)
Anticiper l’impact de fluctuations environnementales de nature climatique ou alimentaire est un enjeu crucial dans les systèmes de productions animales, et plus particulièrement sur la volaille. Cette influence de l’environnement sur les phénotypes passe en partie par des phénomènes épigénétiques, notamment la méthylation de l’ADN, et qui peuvent intervenir dans la régulation de l'expression des gènes. Ce sont des mécanismes qui n'affectent pas la séquence d'ADN mais qui peuvent être transmis par la mitose ou la méiose. Ces interactions entre épigénomes et expression des gènes sont de plus en plus étudiées dans les modèles animaux et chez les plantes. Cependant, les mécanismes de régulation de l'expression du génome par la méthylation de l’ADN sont assez peu connus chez les oiseaux. Ce travail de thèse repose sur deux dispositifs expérimentaux réalisés chez la poule, le but étant de caractériser le méthylome par séquençage haut-débit. Les profils de méthylation le long du génome, et le lien avec l’expression, sont établis d’abord par un séquençage tout-génome (WGBS) au sein d’embryons entiers, puis par un séquençage d'une sous-représentation du génome (RRBS) au sein d’hypothalamus d’individus adultes. À ce jour, aucune étude d'analyses de méthylome par RRBS chez la poule n'a été publiée. Ces deux analyses sont réalisées grâce au développement d'un pipeline bioinformatique, optimisé, disponible à la communauté scientifique. Globalement, le profil de méthylation chez la poule est similaire à ce qui est connu chez les mammifères : les îlots CpG - régions riches en dinucléotides CG, souvent peu méthylées, qui ponctuent le génome principalement dans les régions promotrices des gènes - sont globalement peu méthylés dans les promoteurs sur les données WGBS et RRBS. Les analyses du méthylome des embryons ont confirmé l'absence d'un phénomène de compensation de dose sur les chromosomes sexuels, ou la présence sur le chromosome Z d'une région hyperméthylée. Les analyses des données RRBS révèlent une hyperméthylation globale des CG sur le génome, suggérant une réponse de la méthylation à un stress environnemental. Sur les données WGBS, le niveau de méthylation dans le promoteur est négativement corrélé à l'expression du gène associé. Une méthylation allèle spécifique est également détectée entre les lignées, phénomène mis en évidence pour la première fois chez la poule et dont la fréquence est comparable à ce qui a été observé chez l'Homme. Sur les données RRBS, des résultats préliminaires de la réponse du méthylome aux stress environnementaux montrent le caractère complexe de cette relation. L’utilisation d’aliments moins énergétiques entraînerait une plus grande mobilisation des réserves lipidiques, tandis que les individus soumis à un stress à la chaleur ont un poids corporel plus léger. Une intégration de ces données à des mesures phénotypiques permettrait de faire le lien entre méthylation et environnement. Au-delà de l'aspect fondamental de cette thèse, l'application plus concrète de ces connaissances peut s'appliquer aux systèmes d'élevage pour obtenir des animaux mieux adaptés à l’environnement, en améliorant les caractères de production / Anticipating the impact of environmental changes (on climate and feed) is a crucial issue for livestock production systems, including poultry. The influence of the environment on phenotypes is partly mediated by epigenetic phenomena, including DNA methylation, which may be involved in the regulation of gene expression. These mechanisms do not affect the DNA sequence but can be inherited by mitosis or meiosis. The interactions between epigenomes and gene expression are increasingly being studied in animal models and in plants. However, the mechanisms of regulation of genome expression through DNA methylation are relatively unknown in birds. This thesis work is based on two experimental devices realized in chicken aiming to characterize the methylome by high-throughput sequencing. The methylation patterns across the genome, and their link with expression, were first established by whole-genome bisulfite sequencing (WGBS) in whole embryos, following a reduced representation bisulfite sequencing (RRBS) from hypothalamus of adults. To date, no specific chicken RRBS study has been published. These two analyses were carried out by developing an optimized bioinformatics pipeline, available for scientific community. Overall, the pattern of methylation in chicken is like those in mammals: CpG islands - dinucleotides CG-rich regions which are often poorly methylated, and which are found mainly in the promoter regions of the genome - are generally poorly methylated in promoters on WGBS and RRBS data. Embryo methylome analyses confirmed the absence of a dose-compensation phenomenon on sex chromosomes, or the presence of a hypermethylated region on the Z chromosome. The analyses of RRBS data revealed an overall hypermethylation of CGs across the genome, suggesting a methylation response to environmental stress. From the analysis of WGBS data, we found that the level of methylation in promoters was negatively correlated with the expression of the associated gene. For the first time, a specific allele methylation was also detected between chicken lines whose frequency is comparable to that observed in humans. On the RRBS data, preliminary results of the methylome response to environmental stresses showed the complex nature of this relationship. The use of a low-energy diet would led to greater mobilization of body fat, while individuals with heat stress had a lighter body weight. Integrating these data with phenotypic measurements would allow to link methylation and environment. Beyond the fundamental aspect of this thesis, the method developed in this work could be applied to livestock systems to breed animals better adapted to a changing environment, by improving production traits.
64

The effect of environmental stressors on the immune response to avian infectious bronchitis virus

Lopez, Juan Carlos January 2006 (has links)
The first aim of this research was to determine the prevalence of IBV in broilers within the Canterbury province, New Zealand, in late winter and to search for associations with management or environmental factors. The second aim was to study how ambient stressors affect the immune system in birds, their adaptive capacity to respond, and the price that they have to pay in order to return to homeostasis. In a case control study, binary logistic regression analyses were used to seek associations between the presence of IBV in broilers and various risk factors that had been linked in other studies to the presence of different avian pathogens: ambient ammonia, oxygen, carbon dioxide, humidity and litter humidity. Pairs of sheds were selected from ten large broiler farms in Canterbury. One shed (case) from each pair contained poultry that had a production or health alteration that suggested the presence of IBV and the other was a control shed. Overall, IBV was detected by RT-PCR in 50% of the farms. In 2 of the 5 positive farms (but none of the control sheds) where IBV was detected there were accompanying clinical signs that suggested infectious bronchitis (IB). Ambient humidity was the only risk factor that showed an association (inverse) with the prevalence of IBV (p = 0.05; OR = 0.92). It was concluded within the constraints of the totally enclosed management systems described, that humidity had an influence on the presence of IBV, but temperature, ammonia, carbon dioxide, oxygen or litter humidity had no effect. In another study environmental temperatures were changed in order to affect the biological function and adaptive capacity of chickens following infection with IBV. The 'affective states' of the animal were assessed by measuring levels of corticosterone (CORT) in plasma and tonic immobility (TI). It was found that low (10 +/- 2°C) and high (30 +/- 2°C) temperatures exacerbated the respiratory signs and lesions in birds infected with IBV as compared to those housed at moderate (20 +/- 2°C) temperatures. The chickens housed at high temperatures showed significantly decreased growth, a higher proportion of hepatic lesions (principally haemorrhages) and a longer tonic immobility period, but there was no significant alteration in the plasma levels of CORT. The birds housed at low temperatures developed a higher proportion of heart lesions (hydropericardium, ventricular hypertrophy) and had significantly higher levels of plasma CORT than birds housed under moderate and/or high temperatures. The specific antibody response to IBV decreased in birds housed under high temperatures. Interestingly the birds housed at high temperatures developed significantly higher levels of haemagglutinin antibodies to sheep red blood cells (SRBC) than those birds housed under low or moderated temperatures. Cell mediated immunity was not significantly affected by heat or cold stress in the first 13 days of treatment but at 20 days the levels of interferon gamma in the birds subjected to low temperatures were lower than in the high temperature group. In other trials, the exogenous administration of low physiological doses of oral CORT (as compared to high pharmacological doses typically used in such experiments) to birds resulted in suppression or enhancement of the immune response depending on duration of treatment and/or dose and nature of the antigen. To our knowledge, this is the first study to show that exogenous CORT can produce an enhancement in the immune response in chickens. iv In conclusion, environmental stressors such as high or low temperatures do affect the physiology of the fast-growing broiler. The adjustments the birds have to make to maintain homeostasis impacts on the course of common infectious diseases, such as IB, that normally is mild in the New Zealand poultry industry. The administration of exogenous CORT showed that this hormone may be part of the physiological stress response and acts as a messenger to prepare the immune system for potential challenges (e.g., infection).
65

Amoebae as Hosts and Vectors for Spread of Campylobacter jejuni

Olofsson, Jenny January 2015 (has links)
Campylobacter jejuni is the leading bacterial cause of gastrointestinal diarrheal disease in humans worldwide. This zoonotic pathogen has a complex epidemiology due to its presence in many different host organisms. The overall aim of this thesis was to explore the role of amoebae of the genus Acanthamoeba as an intermediate host and vector for survival and dissemination of C. jejuni. Earlier studies have shown that C. jejuni can enter, survive and replicate within Acanthamoebae spp. In this thesis, I have shown that C. jejuni actively invades Acanthamoeba polyphaga. Once inside, C. jejuni could survive within the amoebae by avoiding localization to degradative lysosomes. We also found that A. polyphaga could protect C. jejuni in acid environments with pH levels far below the range in which the bacterium normally survives. Furthermore, low pH triggered C. jejuni motility and invasion of A. polyphaga. In an applied study I found that A. polyphaga also could increase the survival of C. jejuni in milk and juice both at room temperature and at +4ºC, but not during heating to recommended pasteurization temperatures. In the last study we found that forty environmental C. jejuni isolates with low bacterial concentrations could be successfully enriched using the Acanthamoeba-Campylobacter coculture (ACC) method. Molecular genetic analysis using multilocus sequence typing (MLST) and sequencing of the flaA gene, showed no genetic changes during coculture. The results of this thesis have increased our knowledge on the mechanisms behind C. jejuni invasion and intracellular survival in amoebae of the genus Acanthamoeba. By protecting C. jejuni from acid environments, Acanthamoebae could serve as important reservoirs for C. jejuni e.g. during acid sanitation of chicken stables and possibly as vectors during passage through the stomach of host animals. Furthermore, Acanthamoeba spp. could serve as a vehicle and reservoir introducing and protecting C. jejuni in beverages such as milk and juice. Validation of the ACC method suggests that it is robust and could be used even in outbreak investigations where genetic fingerprints are compared between isolates. In conclusion, Acanthamoeba spp. are good candidates for being natural hosts and vectors of C. jejuni.
66

Influence de stress environnementaux sur les propriétés physicochimiques de jeunes biofilms en cours de formation : étude par spectroscopies vibrationnelles infrarouge-Raman et de force AFM / Influence of environmental stresses on the physico-chemical properties of nascent biofilms during their formation : a vibrational (infrared and Raman) and force (AFM) spectroscopies study

Jamal, Dima 17 June 2015 (has links)
Les biofilms sont des communautés complexes de microorganismes, enchassées dans une matrice auto-secrétée de substances polymériques extracellulaires ou EPS. Les biofilms se forment à la surface de la plupart des matériaux, qu’ils soient de nature biologique ou non, et sont à l’origine de divers problèmes économiques et sanitaires. Les bactéries dans un biofilm, dites bactéries sessiles, présentent en effet des propriétés phénotypiques qui les distinguent de leurs homologues planctoniques, notamment par une résistance accrue aux antibiotiques et aux traitements de désinfection. D’où, la nécessité de prévenir leur formation et/ ou de leur élimination à partir de stratégies mieux adaptées à ce mode de vie en communauté. Le développement de telles stratégies passe entre autre par une meilleure connaissance des contributions physico-chimiques gouvernant les interactions de ces microorganismes avec leur environnement proche notamment lors des étapes initiales de la formation des biofilms. Deux grands objectifs ont été fixés au début de cette thèse, le premier visant à caractériser, in situ et en temps réel la formation de jeunes biofilms de deux modèles bactériens : une souche naturelle et ubiquitaire de Pseudomonas fluorescens et une souche modèle d’Escherichia coli obtenue par génie génétique pour surexprimer un seul type d’EPS. Le deuxième objectif de cette thèse, consiste à étudier leurs réponses à un stress environnemental ou chimique, notamment quand les biofilms doivent se développer dans des conditions extrêmes de pH. Pour atteindre ces objectifs, différentes techniques ont été combinées pour étudier de l’échelle moléculaire à l’échelle cellulaire le développement des biofilms. La spectroscopie Infrarouge à Transformée de Fourier en mode réflexion totale atténuée (FTIR-ATR) a été utilisée pour suivre en temps réel le développement des biofilms. Nous avons pu suivre l’évolution des empreintes spectrales IR-ATR au cours de la formation des biofilms, sous des conditions favorables ou non à leur croissance. De jeunes biofilms de 24 h ont été étudiés par microspectroscopie Raman confocale (MRC), celle ci permettant d’obtenir des informations localisées sur la composition chimique des biofilms. La structure générale des biofilms a été visualisée par la microscopie à épifluorescence. Finalement, les propriétés physico-chimiques des EPS ont été quantifiées par spectroscopie de force atomique à l’échelle de la molécule unique (SMFS pour Single Molecule Force Spectroscopy). / Biofilms are complex communities of microorganisms, embedded in an auto-produced matrix of extracellular polymeric substances or EPS. Biofilms form on the surface of most materials, whether or not they are of biological nature, and cause major economic problems as well as public health concerns. Bacteria within a biofilm also called sessile bacteria, have phenotypic characteristics that distinguish them from their planktonic counterparts, rendering them more resistant to antibiotics and to disinfection strategies. Hence, the prevention of their formation and/ or their elimination requires the use of strategies that are well suited to the sessile mode of life. The development of these strategies begins with a better understanding of the physicochemical contributions that govern the interaction between the sessile community and its environment especially during the first steps of biofilm formation. Two main objectives were defined at the beginning of this thesis, the first was to characterize, in situ, and in real time the development of nascent biofilms. Two bacterial models were studied: a natural and ubiquitous strain of Pseudomonas fluorescens and a model strain of Escherichia coli genetically modified to overexpress one type of EPS. The second objectif was to study their responses towards an environmental or chemical stress; particularly how their development would be affected under extreme conditions of pH. To gain these objectives, different techniques were combined to study from the molecular to the cellular scale the development of biofilms. Fourier Transform Infrared spectroscopy in attenuated total reflection mode was used to evaluate in real time the development of biofilms. We were able to detect changes in the IR-ATR spectral profile along biofilm formation under favorable and non favorable growth conditions. 24 h - old biofilms were characterized using confocal Raman microspectroscopy, which allowed us to gather localized information on their chemical composition. The structure of biofilms was visualized using epifluorescence microscocopy. Finally, physico-chemical properties of EPS were quantified using single molecule force spectroscopy
67

Adaptation de l'Archaea halophile halobacterium salinarum aux stress environnementaux : mécanismes de survie et rôle de la protéolyse intracellulaire / Environnemental stress adaptation of the halophilic Archaea halobaterium salinarum : survival mechanisms and role of intracellular proteolysis.

Marty, Vincent 09 December 2011 (has links)
Les systèmes moléculaires décrits chez les Archaea mettent en évidence un caractère primitif et une simplicité, comparativement à leurs homologues eucaryotes. Par ailleurs, leur caractère extrêmophile a pour conséquence une hyper-robustesse qui rend leur manipulation in vitro et les études structurales beaucoup plus aisées. Ainsi les Archaea représentent de bons modèles pour comprendre les fonctions cellulaires complexes, particulièrement celles qui mettent en jeu de grandes machineries moléculaires, comme celles impliquées dans la protéolyse. Mon travail de thèse a consisté à comprendre les mécanismes de résistance et l'importance des systèmes de protéolyse dans l'adaptation des Archaea halophiles aux stress environnementaux. Les Archaea halophiles accumulent des concentrations multi-molaires de KCl/NaCl dans leur cytosol (3.4M KCl / 1.1M NaCl chez Halobacterium salinarum). Ainsi, les protéines de ces organismes sont elles-mêmes halophiles et ne sont solubles et repliées que dans des conditions de salinité extrêmes (de 2 à 5M).Nous avons étudié la réponse de l'Archaea halophile stricte H. salinarum à des stress, dont les stress à basse salinité, en se focalisant en particulier sur les modifications de la dynamique moléculaire du protéome in vivo (diffusion neutronique). Au cours de ce travail, il a été mis en évidence un phénomène de survie à la basse salinité associé à des modifications morphologiques.Un autre objectif de ma thèse a été de contribuer à la compréhension du rôle dans la protéolyse intracellulaire du complexe aminopeptidasique TET, dans les conditions de stress mises en place dans les études précédentes. / Molecular systems described for Archaea show primitive and simple characteristics, compared to their homologous eukaryotes. In addition, extremophilic characteristic results in an hyper-robust which makes in vitro manipulation and structural studies much easier. Thus, Archaea represent good models for understanding complex cellular functions, particularly those that involve large molecular machines, such as those involved in proteolysis. My thesis consisted in understanding the resistance mechanisms and the importance of proteolytic systems in the adaptation of halophilic Archaea to environmental stresses. Halophilic Archaea accumulate multi-molar concentrations of KCl / NaCl in their cytosol (3.4M KCl / NaCl 1.1M for Halobacterium Salinarum). This requires a very special biochemistry that allows operation in solvents where free water is scarce. Thus, the proteins of these organisms are themselves halophilic and are soluble and folded only in extreme salinity conditions (2 to 5 M). This particular biochemistry partly explain the extraordinary ability of halophilic Archaea to resist physical and chemical stress (temperature, radiation, dehydration). We study the response of the halophilic Archaea strict H. salinarum at low-salinity stress. Indeed, beyond the osmotic shock, the fall of the environment salt concentration causes a decrease in the intracellular KCl concentration, which should have a direct effect on the folding state of intracellular protein, as in case of heat stress. In the first part of this thesis, a study was conduct to determine viability limits and cytosolic modifications, associated with a salinity decrease. These studies involve intracellular salt dosages, viability studies (microscopic counts, color live / dead), induction of chaperone proteins linked to stress response and biophysical neutron experiments, to evaluate the effect of stress on proteins folding. In this work, a phenomenon of survival at low salinity linked to morphological changes was revealed. To describe this phenomenon, this second study involves confocal microscopy experiences, fluorescence microscopy, viability tests, counting on box, scanning electron microscopy, electron microscopy by negative staining, salt intracellular dosages and proteins separation experiments, to study the overall proteome composition during low salinity stress. In this study, a fall of the intracellular K $^+$ concentration and the proteome clarification during stress was revealed. Low salt concentrations causes halophiles proteins denaturation, the accumulation of misfolded proteins in the cytoplasm involves chaperones systems and intracellular proteolysis machinery. In this context, another objective of my thesis was to contribute to the understanding of the intracellular proteolysis role in the PAN-proteasome system and in the aminopeptidase TET complex, in stress conditions established in previous studies. This part of the thesis involves experiments of endopeptidase activity assay, aminopeptidase activity assay, quantification of mRNA genic expression by Northern blot, immunoprecipitation, proteins separation by sucrose gradient and proteasome chemical inhibition (drug). We show the role of the PAN-proteasome system in stress response and we deepen our understanding of the aminopeptidase TET role in vivo. This protease appears to have an independent role of the proteasome complex. The protease TET seems to participate at the amino acids treatment in cells to maintain the metabolic activities in nutritional deficiencies.
68

Varia??o no conte?do prot?ico e pigmentar em variantes crom?ticas de Gracilaria domingensis nas popula??es naturais de Rio do Fogo-RN, BRASIL / Variation in protein and pigment content in colour strains of Gracilaria domingensis in the natural populations of Rio do Fogo (RN)

Pereira, Dinaelza Castelo 23 March 2009 (has links)
Made available in DSpace on 2014-12-17T14:01:58Z (GMT). No. of bitstreams: 1 DinaelzaCP.pdf: 982597 bytes, checksum: 8505a456042640660a4eeec39570774f (MD5) Previous issue date: 2009-03-23 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The seaweed Gracilaria domingensis is a common species in the coast of Rio Grande do Norte. This species lives in the intertidal zone, where colour strains (red, green and brown) co-occur during the whole year. Seaweeds that live in this region are exposed to daily changes and to the rhythm of the tide. During the low tide they are exposed to dissection, hiper-or hipo-osmotic shock, high temperatures and high irradiance. The aim of this study was to analyze whether the pigment and protein content of the colour strains of G. domingensis is affected by some environmental parameters in a temporal scale. The seaweeds were collected during 10 months in the seashore of Rio do Fogo (RN). The total soluble proteins and the phycobiliprotein were extracted in phosphate buffer and the carotenoids were analyzed by a standardized method through HPLC-UV. The pigments analysis showed that phycoerithrin is the most abundant pigment in the three strains. This pigment was strongly correlated with nitrogen and the photosynthetically active radiation. Chlorophyll presented higher concentrations than carotenoids during the whole, but the ratio carotenoid/chlorophyll-a was modified by incident radiation. The most abundant carotenoid was ?-carotene and zeaxanthin, which had higher concentrations in the higher radiation months. The concentration increase of zeaxanthin in this period indicated a photoprotective response of the seaweed. The three strains presented a pigment profile that indicates different radiation tolerance profile. Our results pointed that the green strain is better adapted to high irradiance levels than the red and brown strains / A macroalga Gracilaria domingensis ? comumente encontrada no litoral do Rio Grande do Norte. Esta esp?cie habita a zona intertidal, onde linhagens crom?ticas (vermelha, verde e marrom) co-ocorrem durante todo o ano. Algas que vivem nesta regi?o est?o submetidas a mudan?as diurnas e ao ritmo da mar?. Durante a mar? baixa elas ficam expostas a disseca??o, choque hiper ou hipo-osm?tico, altas temperaturas e elevada irradi?ncia. Este trabalho teve como objetivo geral o estudo do perfil prot?ico e pigmentar das variantes crom?ticas de G. domingensis e a altera??o deste perfil em fun??o dos par?metros ambientais em uma escala temporal. As algas foram coletadas mensalmente, durante 10 meses, nas parias de Rio do Fogo-RN. As prote?nas totais sol?veis e as ficobiliprote?nas foram extra?das em tamp?o fosfato e os caroten?ides foram analisados em um m?todo padronizado por HPLC-UV. A an?lise dos pigmentos mostrou que a ficoeritrina ? o pigmento mais abundante nas tr?s linhagens. Este pigmento esteve fortemente correlacionado com o nitrog?nio e a PAR. Os pigmentos caroten?ides apresentaram concentra??es inferiores as da clorofila-a durante todos os meses, mas a raz?o caroten?ides/clorofila-a foi modificada com o aumento da radia??o. O caroten?ide mais abundante foi o ?-caroteno, seguido da zeaxantina, que esteve em maiores concentra??es nos meses de maior radia??o. O aumento na concentra??o da zeaxantina nesse per?odo indicou uma resposta fotoprotetora da alga. As tr?s linhagens apresentaram um perfil pigmentar que remete a diferentes padr?es de toler?ncia a radia??o. A linhagem verde mostrou ser melhor adaptada a elevados n?veis de irradi?ncias do que a vermelha e a marrom
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Functional characterization of the DELLA RGA-LIKE 3 in Arabidopsis thaliana / Caractérisation fonctionnelle du DELLA RGA-LIKE3 chez Arabidopsis thaliana

Wild, Michael 18 July 2013 (has links)
Les gibbérellines (GA) sont des phytohormones qui régulent divers aspects du développement en réponse aux signaux endogènes et exogènes à la plante. Ainsi face à un stress, les niveaux de GA sont finement contrôlés, permettant une croissance adaptée aux contraintes environnementales. Au niveau moléculaire, les GA stimulent la croissance de la plante en s’opposant aux protéines DELLAs (DELLAs), facteurs nucléaires qui inhibent la croissance. Les DELLAs présentent plusieurs caractéristiques fonctionnelles notables, une activité transactivatrice et la capacité d’interagir avec d’autres protéines régulatrices, comme les répresseurs de la signalisation Jasmonate (JA), JA ZIM-domain (JAZ). Le génome d’Arabidopsis thaliana code pour cinq DELLAs possédant des fonctions redondantes et spécifiques. Le but de mon travail de thèse a été la caractérisation de la fonction biologique d’une DELLA, RGA-LIKE3 (RGL3). J’ai pu montrer que RGL3 modifie la défense de la plante face à des stresses biotiques. / The phytohormones gibberellins (GA) regulate major aspects of plant growth in response to endogenous and environmental signals. Upon the perception of stress, the levels of bioactive GA are adjusted, hence allowing a flexible growth response to environmental variability. At a molecular level, GA promote growth by stimulating the degradation of the growth repressing DELLA proteins. DELLAs are versatile nuclear proteins with several remarkable features, such as transactivation activity and protein–protein interaction capacities. Thus DELLAs interact with a series of highly divergent proteins, including different transcription factor families, but also the Jasmonate (JA) ZIM-domain (JAZ) proteins, repressors of JA signaling. The aim of this thesis work consisted in the characterization of the biological function of the DELLA RGA-LIKE3. I could show that RGL3 modulates plant defense responses against biotic stresses.
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Como a proximidade de atividades agrícolas afeta libélulas? Efeitos sobre traços biológicos de Hetaerina rosea / How to proximity of agricultural activities affects dragonflies? Effects on biological traits of Hetaerina rosea

Pinto, Nelson Silva 27 February 2014 (has links)
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No. of bitstreams: 2 Dissertação - Nelson Silva Pinto - 2014.pdf: 1321543 bytes, checksum: eeca73fc90de8b75b567ca0300d5b598 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-02-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / In our study, we evaluated the proximity of agricultural activities and the percentage of area with anthropogenic activities affect the development of Hetaerina rosea. The impacts of human activities may lead to instability of the development of organisms before they affect population variables. Understanding how these effects are negative for individuals can be a powerful metric and assist in decision making regarding the management of impacts. The most commonly used measure to assess how the developmental instability is fluctuating asymmetry (AF). Although several studies provide support for using AF as an indicator of stress during development, others have questioned its validity as a bioindication tool. Our findings suggest little support to AF. However, we must consider that development is an integrated process and other biological traits may be affected during development. Weight is one of these variables. In our study, we observed that individuals from environments near to agricultural activities had lower weights. This result can be explained by the influence of stresses during the larval stage, such as, for example, the presence of pesticides in streams, increased competition from adult males by breeding territories or lesser quantity/quality of food availability in streams nearby to agricultural activities. Although the hypothesis of food stress in the larval stage or adult stage cannot be considered separately, they may provide that, in Odonata, the size would be maintained in detriment of weight, since stresses during the larval stage lead to energy investment in compensatory growth through increased metabolism. Our results can be interpreted according to this premise since no significant differences between the average size of the wings. Thus, we suggest that for a more integrated view of how anthropogenic stresses affect the development of Odonata other variables should be used in addition to AF, such as weight and size. / Em nosso estudo avaliamos como a proximidade de atividades agrícolas e a porcentagem de área com atividades antrópicas afetam o desenvolvimento de Hetaerina rosea. Os impactos de atividades humanas podem levar à instabilidade do desenvolvimento dos organismos muito antes de afetar variáveis populacionais. A compreensão de como estes efeitos são negativos para os indivíduos pode ser uma métrica poderosa e auxiliar na tomada de decisão com relação ao manejo de impactos. A medida mais comumente utilizada como para avaliar a instabilidade do desenvolvimento é a assimetria flutuante (AF). Embora diversos estudos ofereçam suporte ao uso de AF como indicador de estresses durante o desenvolvimento, outros questionam sua validade como ferramenta para bioindicação. Nossos resultados sugerem pouco suporte à AF. Entretanto, devemos considerar que o desenvolvimento é um processo integrado e que outros traços biológicos podem ser afetados durante o desenvolvimento. O peso é uma destas variáveis. Em nosso estudo, observamos que os indivíduos provenientes de ambientes próximos à atividades agrícolas apresentaram pesos menores. Esse resultado pode ser explicado pela influência de estresses durante o estágio larval, tais como, por exemplo, a presença de agroquímicos nos riachos onde se desenvolveram, maior competição dos machos adultos por territórios de reprodução ou menor quantidade/qualidade de alimento disponível nos riachos próximos à atividades agrícolas. Embora as hipóteses de estresse alimentar na fase larval ou na fase adulta não possam ser consideradas separadamente, elas podem prever que, em Odonata, o tamanho seria mantido, no entanto, o peso não, uma vez que estresses durante o estágio larval levam ao investimento de energia no crescimento compensatório através de aumento do metabolismo. Nossos resultados podem ser interpretados de acordo com essa premissa uma vez que não observamos diferenças significativas entre o tamanho médio das asas. Desta maneira, sugerimos que para obter uma visão mais integrada de como estresses de origem antropogênica afetam o desenvolvimento de Odonata outras variáveis devem ser utilizadas além da AF, tais como o peso e o tamanho.

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