Engineered nanomaterials (ENMs) have become ubiquitous in consumer products and industrial applications, and consequently the environment. Much of the environmentally released ENMs are expected to enter terrestrial ecosystems via land application of nano-enriched biosolids to agricultural fields. Among the organisms most likely to encounter nano-enriched biosolids are the key soil bacteria known as plant growth promoting rhizobacteria (PGPR). I reviewed what is known concerning the toxicological effects of ENMs to PGPR and observed the need for high-throughput methods to evaluate lethal and sublethal toxic responses of aerobic microbes. I addressed this issue by developing high-throughput microplate assays which allowed me to normalize oxygen consumption responses to viable cell estimates. Oxygen consumption is a crucial step in cellular respiration which may be examined relatively easily along with viability and may provide insight into the metabolic/physiological response of bacteria to toxic substances. Because many of the most toxic nanomaterials (i.e. metal containing materials) exhibit some level of ionic dissolution, I first developed my methods by examining metal ion responses in the PGPR, Bacillus amyloliquefaciens GB03. I found this bacterium exhibits differential oxygen consumption responses to Ag+, Zn2+, and Ni2+. Exposure to Ag+ elicited pronounced increases in O2 consumption, particularly when few viable cells were observed. Also, while Ni2+ and Zn2+ are generally thought to induce similar toxic responses, I found O2 consumption per viable cell was much more variable during Ni2+ exposure and that Zn2+ induced increased O2 utilization to a lesser extent than Ag+. Additionally, I showed my method is useful for probing toxicity of traditional antibiotics by observing large increases in O2 utilization in response to streptomycin, which was used as a positive control due to its known effects on bacterial respiration. After showing the utility of my method for examining metal ion responses in a single species of PGPR, I investigated the toxicity of silver ENMs (AgENMs) and ions to three PGPR, B. amyloliquefaciens GB03, Sinorhizobium meliloti 2011, and Pseudomonas putida UW4. The ENM exposures consisted of untransformed, polyvinylpyrrolidone coated silver ENMs (PVP-AgENMs) and 100% sulfidized silver ENMs (sAgENMs), which are representative of environmentally transformed AgENMs. I observed species specific O2 consumption responses to silver ions and PVP-AgENMs. Specifically, P. putida exhibited increased O2 consumption across the observed range of viable cells, while B. amyloliquefaciens exhibited responses similar to those found in my first study. Additionally, S. meliloti exhibited more complex responses to Ag+ and PVP-AgENMs, with decreased O2 consumption when cell viability was ~50-75% of no metal controls and increased O2 consumption when cell viability was <50%. I also found the abiotically dissolved fraction of the PVP-AgENMs was likely responsible for most of the toxic response, while abiotic dissolution did not explain the toxicity of sAgENMs. My work has yielded a straightforward, cost-effective, and high-throughput method of evaluating viability and oxygen consumption in aerobic bacteria. I have used this method to test a broad range of toxic substances, including, metal ions, antibiotics, and untransformed and transformed ENMs. I observed species specific toxic responses to Ag+, PVP-AgENMs, and sAgENMs in PGPR. These results not only show the clear utility of the methodology, but also that it will be crucial to continue examining the responses of specific bacterial strains even as nanotoxicology, as a field, must move toward more complex and environmentally relevant systems.

Problématique : Le concept d’« Hôpital promoteur de santé » (HPS) a émergé dans le sillon de la Charte d’Ottawa (1986) qui plaide notamment pour une réorientation des services de santé vers des services plus promoteurs de santé. Il cible la santé des patients, du personnel, de la communauté et de l’organisation elle-même. Dans le cadre de la réforme du système de santé au Québec qui vise à rapprocher les services de la population et à faciliter le cheminement de toute personne au sein d’un réseau local de services de santé et de services sociaux (RLS), l’adoption du concept HPS semble constituer une fenêtre d’opportunité pour les CHU, désormais inclus dans des réseaux universitaires intégrés de soins de santé et rattachés aux RLS, pour opérer des changements organisationnels majeurs. Face au peu de données scientifiques sur l’implantation des dimensions des projets HPS, les établissements de santé ont besoin d’être accompagnés dans ce processus par le développement de stratégies claires et d’outils concrets pour soutenir l’implantation. Notre étude porte sur le premier CHU à Montréal qui a décidé d’adopter le concept et d’implanter notamment un projet pilote HPS au sein de son centre périnatal. Objectifs : Les objectifs de la thèse sont 1) d’analyser la théorie d’intervention du projet HPS au sein du centre périnatal; 2) d’analyser l’implantation du projet HPS et; 3) d’explorer l’intérêt de l’évaluation développementale pour appuyer le processus d’implantation. Méthodologie : Pour mieux comprendre l’implantation du projet HPS, nous avons opté pour une étude de cas qualitative. Nous avons d’abord analysé la théorie d’intervention, en procédant à une revue de la littérature dans le but d’identifier les caractéristiques du projet HPS ainsi que les conditions nécessaires à son implantation. En ce qui concerne l’analyse d’implantation, notre étude de cas unique a intégré deux démarches méthodologiques : l’une visant à apprécier le niveau d’implantation et l’autre, à analyser les facteurs facilitants et les contraintes. Enfin, nous avons exploré l’intérêt d’une évaluation développementale pour appuyer le processus d’implantation. À partir d’un échantillonnage par choix raisonnés, les données de l’étude de cas ont été collectées auprès d’informateurs clés, des promoteurs du projet HPS, des gestionnaires, des professionnels et de couples de patients directement concernés par l’implantation du projet HPS au centre périnatal. Une analyse des documents de projet a été effectuée et nous avons procédé à une observation participante dans le milieu. Résultats : Le premier article sur l’analyse logique présente les forces et les faiblesses de la mise en oeuvre du projet HPS au centre périnatal et offre une meilleure compréhension des facteurs susceptibles d’influencer l’implantation. Le second article apprécie le niveau d’implantation des quatre dimensions du projet HPS. Grâce à la complémentarité des différentes sources utilisées, nous avons réussi à cerner les réussites globales, les activités partiellement implantées ou en cours d’implantation et les activités reposant sur une théorie d’intervention inadéquate. Le troisième article met en évidence l’influence des caractéristiques de l’intervention, des contextes externe et interne, des caractéristiques individuelles sur le processus d’implantation à partir du cadre d’analyse de l’implantation développé par Damschroder et al. (2009). Enfin, le dernier article présente les défis rencontrés par la chercheure dans sa tentative d’utilisation de l’évaluation développementale et propose des solutions permettant d’anticiper les difficultés liées à l’intégration des exigences de recherche et d’utilisation. Conclusion : Cette thèse contribue à enrichir la compréhension de l’implantation du projet HPS dans les établissements de santé et, particulièrement, en contexte périnatal. Les résultats obtenus sont intéressants pour les chercheurs et les gestionnaires d’hôpitaux ou d’établissements de santé qui souhaitent implanter ou évaluer les projets HPS dans leurs milieux. / Problem: The concept of health promoting hospitals (HPH) emerged in the wake of the 1986 Ottawa Charter, which notably calls for the reorientation of health services toward more health promoting services. The concept targets the health of patients, staff, the community and the organization itself. In the context of Quebec’s health system reform that aims to bring services closer to the population and facilitates the journey of any person within a local network of health and social services (RLS), the adoption of the HPH concept appears to be a window of opportunity for teaching hospitals, now included in integrated university health care networks (RUIS) and linked to RLS, to carry out major organizational changes. Given the paucity of evidence regarding implementation of the different dimensions of HPH projects, there is a need for health care facilities to be accompanied in this process through the development of clear-cut strategies and practical tools in order to support implementation. Our study will focus on the first university hospital in Montreal that decided to adopt the concept and implement a HPH pilot project within its perinatal centre. Objectives: The objectives of this thesis are 1) to analyse the intervention theory underlying the HPH project; 2) to analyse the implementation of the HPH project; and 3) to explore the potential of developmental evaluation as a means of supporting the implementation process. Methods: To better understand the implementation of the HPS project, we opted for a qualitative case study. We first analyzed the theory of intervention by conducting a literature review in order to identify the characteristics of an HPS project as well as the conditions necessary for its implementation. Then, as regards the implementation analysis, our single case study had two methodological approaches: one to assess the level of implementation and the other to analyze facilitators and barriers to implementation. Finally, we explored the interest of using developmental evaluation to support the implementation process. Based on purposeful sampling, data of the case study were collected from key informants, HPH project promoters, managers, professionals and couples of patients directly concerned by the HPH project implementation in the perinatal centre. A documentary analysis of project documents has been done and participant observation was conducted in the setting. Results: The first article is about a logical analysis that presents the strengths and weaknesses of the implementation of the HPH project within the birthing centre, and offers a deeper comprehension of the factors likely to influence implementation. The second article assesses the level of implementation of the four dimensions of the HPH project. The complementarity of the different sources used allowed us to identify global successes, activities that were implemented only partially or still in the process of implementation, and activities that were based on an inadequate intervention theory. The third article shows the influence of intervention characteristics, outer and inner settings, and individual characteristics upon the implementation process, using the consolidated framework for implementation research developed by Damschroder et al. (2009). The fourth and final article highlights the challenges encountered by the researcher in attempting to use developmental evaluation, and puts forth solutions that will enable researchers to anticipate the difficulties that can emerge from integrating the demands of research and of utilization. Conclusion: This thesis contributes to the understanding of HPH project implementation in health care facilities, particularly within a perinatal care context. The results we have obtained are of interest to researchers as well as hospital or health care facility administrators who wish to implement or evaluate HPH projects.

Paper One: Social and emotional literacy has become an educational agenda on a national and international level. Schools universally are addressing deviant behaviour through a social learning perspective. Emotional literacy (EL) reflects three of the five outcomes for “Every Child Matters: Change for Children” (DfES, 2003): to be healthy, to enjoy and achieve and to make a positive contribution. EI is also embedded in the legal framework for the associated reform that is set out in the Children Act (2004). The five outcomes for Every Child Matters are statutory demands on educational institutions and welfare services. Since EL is reflected in the outcomes, it suggests that it too needs to be addressed. However, schools have the flexibility in how they chose to address it. This study aims to evaluate a personalised approach in promoting EL in Key Stage Two (KS2) children in one primary school. A pre and post design was used to evaluate the school’s new approach (NA). Class teachers completed Strengths and Difficulties Questionnaires (SDQs) for 75 children pre-NA and post-NA. Statistical tests were used to: 1) Compare the pre SDQ scores to the post SDQ scores to determine whether the NA produced a significant change. 2) Determine where there was a significant difference between the research sample scores and the expected value scores according to the classification of the SDQ scores. 3) Compare the research sample SDQ scores to the SDQ scores from the norm data of the British population. Results of the statistical analysis suggest that the NA was effective as there was a significant improvement in the overall general behaviour according to the total difficulties scores. The statistical analysis revealed mixed results for the five scale scores. The hyperactivity scale, the peer problems scale and the emotional symptoms scale showed significant improvements. However, the pro social scale showed a significant decline and the conduct problems scale was the only scale that showed no significant difference between pre-NA and post- NA. Further tests conducted to strengthen the quality of the sample showed the pro social scale was in line with the British norms even with the significant decline. Similarly, the conduct problems scale was in line with the British norm post-NA. The research has produced encouraging statistics for the effectiveness of NA on the children’s behaviour, however, it calls for a re-evaluation of the NA in order to improve the pro social behaviour and lessen conduct problems. Section One Introduction 1.1 Purpose Personal, social and emotional development is as much a concern as academic development in children. Research suggests that social and emotional skills are needed to succeed in school (Thompson, 2002) to establish and sustain relationships, reduce aggressive behaviour (Nagin & Tremblay, 1999) and create an ideal learning and teaching environment. The objective of this research is to determine whether the school’s personalised EL programme (NA) produces favourable behavioural outcomes in children. Paper Two: Social and emotional literacy has become an educational agenda on a national and international level. Schools universally are addressing deviant behaviour through a social learning perspective. Emotional literacy (EL) reflects three of the five outcomes for “Every Child Matters: Change for Children” (DfES, 2003): to be healthy, to enjoy and achieve and to make a positive contribution. EI is also embedded in the legal framework for the associated reform that is set out in the Children Act (2004). The five outcomes for Every Child Matters are statutory demands on educational institutions and welfare services. Since EL is reflected in the outcomes, it suggests that it too needs to be addressed. However, schools have the flexibility in how they chose to address it. This study aims to evaluate a personalised approach in promoting EL in Key Stage Two (KS2) children in one primary school. A pre and post design was used to evaluate the school’s new approach (NA). Class teachers completed Strengths and Difficulties Questionnaires (SDQs) for 75 children pre-NA and post-NA. Statistical tests were used to: 1) Compare the pre SDQ scores to the post SDQ scores to determine whether the NA produced a significant change. 2) Determine where there was a significant difference between the research sample scores and the expected value scores according to the classification of the SDQ scores. 3) Compare the research sample SDQ scores to the SDQ scores from the norm data of the British population. Results of the statistical analysis suggest that the NA was effective as there was a significant improvement in the overall general behaviour according to the total difficulties scores. The statistical analysis revealed mixed results for the five scale scores. The hyperactivity scale, the peer problems scale and the emotional symptoms scale showed significant improvements. However, the pro social scale showed a significant decline and the conduct problems scale was the only scale that showed no significant difference between pre-NA and post- NA. Further tests conducted to strengthen the quality of the sample showed the pro social scale was in line with the British norms even with the significant decline. Similarly, the conduct problems scale was in line with the British norm post-NA. The research has produced encouraging statistics for the effectiveness of NA on the children’s behaviour, however, it calls for a re-evaluation of the NA in order to improve the pro social behaviour and lessen conduct problems.

La régulation post-transcriptionnelle joue un rôle de premier plan dans le contrôle fin de l’expression génique en permettant une modulation de la synthèse de protéines dans le temps et l’espace, en fonction des besoins de la cellule. Ainsi, des protéines reconnaissant des éléments d’ARN présents sur des transcrits peuvent influencer toutes les étapes de leur existence, soit leur épissage, leur export nucléaire, leur localisation subcellulaire, leur traduction et leur dégradation. Staufen1 (Stau1) est un membre de la famille des protéines liant l’ARN double-brin qui contribue à la régulation post-transcriptionnelle par son implication dans des mécanismes qui vont promouvoir l’épissage alternatif, le transport, la dé-répression de la traduction et l’induction de la dégradation d’ARN messagers (ARNm) spécifiques. L’identité des cibles potentielles de Stau1 est maintenant connue puisqu’une étude à l’échelle du génome a montré que la protéine s’associe à près de 7% du transcriptome des cellules HEK293T. Ces ARNm se classent dans un large éventail de catégories fonctionnelles, mais il est tout de même intéressant de noter qu’une grande proportion d’entre eux code pour des protéines reliées au métabolisme cellulaire et à la régulation de processus cellulaires. En considérant toutes ces informations, nous avons émis l’hypothèse que les différentes activités de Stau1 puissent être modulées afin de contrôler adéquatement l’expression des transcrits liés par la protéine. Dans la mesure où certains ARNm faisant partie des complexes définis par la présence de Stau1 codent pour des régulateurs clés de la prolifération cellulaire, nous avons voulu examiner si l’expression de la protéine varie au cours du cycle de division cellulaire. Nous avons montré que l’abondance de Stau1 est maximale en début de mitose et qu’elle diminue ensuite lorsque les cellules complètent la division cellulaire. Nous avons ensuite découvert que cette baisse d’expression de Stau1 en sortie de mitose dépend du complexe promoteur d’anaphase/cyclosome (APC/C). En soutien à l’idée que Stau1 soit une cible de cette ubiquitine ligase de type E3, nous avons de plus démontré que Stau1 est ubiquitiné et dégradé par le protéasome. Ce contrôle des niveaux de Stau1 semble important puisque la surexpression de la protéine retarde la sortie de mitose et entraîne une diminution importante de la prolifération cellulaire. Par ailleurs, nous avons supposé que les différentes fonctions de Stau1 puissent également être sujettes à une régulation. Compte tenu que les activités de nombreuses protéines liant l’ARN peuvent être contrôlées par des modifications post-traductionnelles telles que la phosphorylation, nous avons voulu tester la possibilité que Stau1 soit phosphorylé. L’immunopurification de Stau1 et son analyse par spectrométrie de masse nous a permis d’identifier trois phosphosites dans la protéine. L’évaluation du rôle de ces événements de phosphorylation à l’aide de mutants phoshomimétiques ou non-phoshorylables a révélé que la modification de Stau1 pourrait compromettre son association à la protéine UPF1. Comme cette interaction est nécessaire pour déstabiliser les transcrits liés par Stau1, nos résultats suggèrent fortement que la fonction de Stau1 dans la dégradation d’ARNm est régulée négativement par sa phosphorylation. Toutes ces données mettent en lumière l’importance des modifications post-traductionnelles telles que l’ubiquitination et la phosphorylation dans la modulation de l’expression et des fonctions de Stau 1. Somme toute, il est vraisemblable que ces mécanismes de contrôle puissent avoir un impact significatif sur le destin des ARNm liés par Stau1, particulièrement dans un contexte de progression dans le cycle cellulaire. / Post-transcriptional regulation plays a major role in the fine tuning of gene expression by allowing a modulation of protein synthesis in space and time, according to cellular requirements. For instance, proteins recognizing RNA elements on transcripts can influence all the steps of their existence, such as their splicing, nuclear export, subcellular localization, translation and degradation. Staufen1 (Stau1) is a member of the double-stranded RNA-binding protein family that contributes to the post-transcriptional regulation of gene expression by its involvement in mechanisms that promote alternative splicing, transport, de-repression of translation and decay of specific messenger RNAs (mRNAs). The identity of potential Stau1 targets is now known as genome-wide analyses have shown that the protein is associated with about 7% of the HEK293T cell transcriptome. Although these mRNAs are classified in a broad range of functional categories, a large proportion of them code for proteins related to cellular metabolism and regulation of cellular processes. Considering all this information, we hypothesized that the different activities of Stau1 may be modulated in order to control appropriately the expression of Stau1-bound mRNAs. Since some of the mRNAs that are part of Stau1-containing complexes encode key regulators of cell proliferation, we wanted to examine whether Stau1 expression fluctuates during the cell division cycle. We showed that Stau1 abundance peaks at the onset of mitosis and then decreases as cells complete division. We then found that Stau1 down-regulation in mitosis exit is mediated by the anaphase promoting complex/cyclosome (APC/C). To support the idea that Stau1 is a target of this E3-ubiquitin ligase, we further demonstrated that Stau1 is ubiquitinated and degraded by the proteasome. The importance of controlling Stau1 levels during the cell cycle is underscored by the observation that its overexpression delays mitotic exit and impairs cell proliferation. Furthermore, we speculated that Stau1 different functions may also be regulated. In the view that the activities of numerous RNA-binding proteins can be controlled by post-translational modifications such as phosphorylation, we tested the possibility that Stau1 is phosphorylated. Mass spectrometry analysis of immunopurified Stau1 allowed the identification of three phosphosites in this protein. Assessment of the role of these phosphorylation events using phosphomimetic or non-phosphorylatable mutants revealed that Stau1 phosphorylation may compromise its association with Upf1. Because this interaction is necessary to elicit the destabilisation of Stau1-bound RNAs, our results strongly suggest that Stau1 function in mRNA decay is negatively regulated by its phosphorylation. Collectively, these data highlight the importance of post-translational modifications such as ubiquitination and phosphorylation in the modulation of Stau1 expression and functions. Overall, the mechanisms that control Stau1 are likely to have a significant impact on the fate of Stau1-bound mRNAs, especially in the context of cell cycle progression.

Problématique : Le concept d’« Hôpital promoteur de santé » (HPS) a émergé dans le sillon de la Charte d’Ottawa (1986) qui plaide notamment pour une réorientation des services de santé vers des services plus promoteurs de santé. Il cible la santé des patients, du personnel, de la communauté et de l’organisation elle-même. Dans le cadre de la réforme du système de santé au Québec qui vise à rapprocher les services de la population et à faciliter le cheminement de toute personne au sein d’un réseau local de services de santé et de services sociaux (RLS), l’adoption du concept HPS semble constituer une fenêtre d’opportunité pour les CHU, désormais inclus dans des réseaux universitaires intégrés de soins de santé et rattachés aux RLS, pour opérer des changements organisationnels majeurs. Face au peu de données scientifiques sur l’implantation des dimensions des projets HPS, les établissements de santé ont besoin d’être accompagnés dans ce processus par le développement de stratégies claires et d’outils concrets pour soutenir l’implantation. Notre étude porte sur le premier CHU à Montréal qui a décidé d’adopter le concept et d’implanter notamment un projet pilote HPS au sein de son centre périnatal. Objectifs : Les objectifs de la thèse sont 1) d’analyser la théorie d’intervention du projet HPS au sein du centre périnatal; 2) d’analyser l’implantation du projet HPS et; 3) d’explorer l’intérêt de l’évaluation développementale pour appuyer le processus d’implantation. Méthodologie : Pour mieux comprendre l’implantation du projet HPS, nous avons opté pour une étude de cas qualitative. Nous avons d’abord analysé la théorie d’intervention, en procédant à une revue de la littérature dans le but d’identifier les caractéristiques du projet HPS ainsi que les conditions nécessaires à son implantation. En ce qui concerne l’analyse d’implantation, notre étude de cas unique a intégré deux démarches méthodologiques : l’une visant à apprécier le niveau d’implantation et l’autre, à analyser les facteurs facilitants et les contraintes. Enfin, nous avons exploré l’intérêt d’une évaluation développementale pour appuyer le processus d’implantation. À partir d’un échantillonnage par choix raisonnés, les données de l’étude de cas ont été collectées auprès d’informateurs clés, des promoteurs du projet HPS, des gestionnaires, des professionnels et de couples de patients directement concernés par l’implantation du projet HPS au centre périnatal. Une analyse des documents de projet a été effectuée et nous avons procédé à une observation participante dans le milieu. Résultats : Le premier article sur l’analyse logique présente les forces et les faiblesses de la mise en oeuvre du projet HPS au centre périnatal et offre une meilleure compréhension des facteurs susceptibles d’influencer l’implantation. Le second article apprécie le niveau d’implantation des quatre dimensions du projet HPS. Grâce à la complémentarité des différentes sources utilisées, nous avons réussi à cerner les réussites globales, les activités partiellement implantées ou en cours d’implantation et les activités reposant sur une théorie d’intervention inadéquate. Le troisième article met en évidence l’influence des caractéristiques de l’intervention, des contextes externe et interne, des caractéristiques individuelles sur le processus d’implantation à partir du cadre d’analyse de l’implantation développé par Damschroder et al. (2009). Enfin, le dernier article présente les défis rencontrés par la chercheure dans sa tentative d’utilisation de l’évaluation développementale et propose des solutions permettant d’anticiper les difficultés liées à l’intégration des exigences de recherche et d’utilisation. Conclusion : Cette thèse contribue à enrichir la compréhension de l’implantation du projet HPS dans les établissements de santé et, particulièrement, en contexte périnatal. Les résultats obtenus sont intéressants pour les chercheurs et les gestionnaires d’hôpitaux ou d’établissements de santé qui souhaitent implanter ou évaluer les projets HPS dans leurs milieux. / Problem: The concept of health promoting hospitals (HPH) emerged in the wake of the 1986 Ottawa Charter, which notably calls for the reorientation of health services toward more health promoting services. The concept targets the health of patients, staff, the community and the organization itself. In the context of Quebec’s health system reform that aims to bring services closer to the population and facilitates the journey of any person within a local network of health and social services (RLS), the adoption of the HPH concept appears to be a window of opportunity for teaching hospitals, now included in integrated university health care networks (RUIS) and linked to RLS, to carry out major organizational changes. Given the paucity of evidence regarding implementation of the different dimensions of HPH projects, there is a need for health care facilities to be accompanied in this process through the development of clear-cut strategies and practical tools in order to support implementation. Our study will focus on the first university hospital in Montreal that decided to adopt the concept and implement a HPH pilot project within its perinatal centre. Objectives: The objectives of this thesis are 1) to analyse the intervention theory underlying the HPH project; 2) to analyse the implementation of the HPH project; and 3) to explore the potential of developmental evaluation as a means of supporting the implementation process. Methods: To better understand the implementation of the HPS project, we opted for a qualitative case study. We first analyzed the theory of intervention by conducting a literature review in order to identify the characteristics of an HPS project as well as the conditions necessary for its implementation. Then, as regards the implementation analysis, our single case study had two methodological approaches: one to assess the level of implementation and the other to analyze facilitators and barriers to implementation. Finally, we explored the interest of using developmental evaluation to support the implementation process. Based on purposeful sampling, data of the case study were collected from key informants, HPH project promoters, managers, professionals and couples of patients directly concerned by the HPH project implementation in the perinatal centre. A documentary analysis of project documents has been done and participant observation was conducted in the setting. Results: The first article is about a logical analysis that presents the strengths and weaknesses of the implementation of the HPH project within the birthing centre, and offers a deeper comprehension of the factors likely to influence implementation. The second article assesses the level of implementation of the four dimensions of the HPH project. The complementarity of the different sources used allowed us to identify global successes, activities that were implemented only partially or still in the process of implementation, and activities that were based on an inadequate intervention theory. The third article shows the influence of intervention characteristics, outer and inner settings, and individual characteristics upon the implementation process, using the consolidated framework for implementation research developed by Damschroder et al. (2009). The fourth and final article highlights the challenges encountered by the researcher in attempting to use developmental evaluation, and puts forth solutions that will enable researchers to anticipate the difficulties that can emerge from integrating the demands of research and of utilization. Conclusion: This thesis contributes to the understanding of HPH project implementation in health care facilities, particularly within a perinatal care context. The results we have obtained are of interest to researchers as well as hospital or health care facility administrators who wish to implement or evaluate HPH projects.

The topic of this thesis is to use descant recorders (hereinafter recorder) as a therapeutic agent in children with behavioral difficulties. The main aim of the study is to analyze the impact of activities with the recorder on the problem phenomena in the behavior of selected individuals and further explore the views of preschool teachers of their own readiness to manage problem behavior in their classrooms. The theoretical part is mainly engaged in the therapeutic use of recorders and characteristics of problematic behavior. Considerable space is devoted to the review of options to promote desirable behavior in pre school children and areas of music therapy. With regard to the desired objectives mixed research is chosen in the practical part. Using a questionnaire opinions of preschool teachers on the issue of educational difficulties in children are collected and also the possibility to use the recorder for therapeutic purposes. The case studies demonstrate the impact of the recorder on the manifestations of problematic behavior. The conclusion summarizes the findings of the study. KEYWORDS descant recorder, manifestations of problematic behavior, behavior disorders, promoting the desired behavior, music therapy, supporting procedures, morning circle, therapeutic - formative methods

Die vorliegende Arbeit behandelte den Einfluss von membran-assoziierten Faktoren im Hippocampus auf das axonale Wachstum, zum einen während der Entwicklung des entorhino-hippocampalen Systems und zum anderen nach Deafferenzierung des adulten Hippocampus. Mit Hilfe des Streifenassays und des Längenauswachsassays wurden zuerst die maturationsabhängigen Eigenschaften von membran-assoziierten Faktoren im Hippocampus getestet. Es zeigte sich, dass entorhinale Axone zwischen ihrem normotypischen Zielgebiet und Kontrollregionen diskrimieren können und bevorzugt auf Membranen ihres Zielgebiets wachsen. Im Folgenden wurden dann Axonen hippocampale Membranen unterschiedlicher Entwicklungsstadien im Streifenassay angeboten. In diesem experimentellen Ansatz wuchsen entorhinale Axone präferenziell auf jenen hippocampalen Membranen, die aus dem Entwicklungsstadium stammen, in den die entorhinalen Fasern in vivo in den Hippocampus einwachsen. Diese Experimente ergaben, dass das in vivo zeitlich genau regulierte Einwachsen entorhinaler Fasern in den Hippocampus von membran-assoziierten Faktoren determiniert ist und ein Zeitfenster für das Vorhandensein dieser Faktoren im Hippocampus existiert. Eines der wesentlichen Charakteristika der Maturation des zentralen Nervensystem ist die Bildung von Myelin und die Myelinisierung von Fasertrakten. Immunozytochemische Analysen mit Myelin-spezifischen Markern ergaben, dass dieses maturationsabhängige Auswachsverhalten zeitlich gut mit der Myelinisierung dieser Hirnregion korreliert. Eine Reihe von in vivo und in vitro Experimenten verschiedener Arbeitsgruppen demonstrierten, dass Myelin starke auswachsinhibitorische Eigenschaften hat, die sogar den Kollaps von Wachstumskolben induzieren können. In Längenauswachsassays zeigte sich, dass Myelin einen starken inhibitorischen Effekt auf das Längenwachstum von entorhinalen Axonen hat. Mit physikalischen Separationstechniken und unter Verwendung des funktionellen Antikörpers gegen inhibitorische Myelinproteine (IN-1) konnte dieser Effekt neutralisiert werden und das neuronale Längenwachstum war wieder vergleichbar zur Kontrollsituation. Untersuchungen im Streifenassay ergaben zusätzlich, dass das wachstumsinhibitorische Myelin und seine Komponenten keine axonalen Lenkungseigenschaften hatte und die gerichtete Zielfindung axonalen Auswachsens nicht beeinflusst. In weiteren Experimenten wurden die membran-assoziierten Faktoren im deafferenzierten Hippocampus untersucht. Dabei zeigte sich, dass nach einer Läsion wachstumsfördernde Faktoren in hippocampalen Membranen vorliegen. Zusätzlich liegen in einem engen Zeitfenster axonale Lenkungsmoleküle vor mit vergleichbarer Attraktivität für entorhinale Axone, wie sie aus entsprechenden Entwicklungsstadien bekannt sind. Die Experimente lassen den Schluss zu, dass im deafferenzierten Hippocampus Faktoren läsionsinduziert werden, und dass diese Faktoren membran-assoziiert sind. Es wird seit langem angenommen, dass das ZNS von adulten Vertebraten in seinem zellulären Zustand determiniert ist und zu keinen grösseren plastischen Veränderungen fähig ist. Gerade nach einer Schädigung von adultem ZNS ist die Regenerationsfähigkeit im Unterschied zu jungen, postnatalen ZNS sehr eingeschränkt. Die beschränkte Regenerationsfähigkeit des adulten ZNS ist wesentlich determiniert durch die Präsenz des auswachsinhibitorischen Myelins. Nichtsdestotrotz gibt es kompensatorisches Sprouting im Hippocampus, die verlorengegangene synaptische Kontakte ersetzen. Die Identifizierung der Faktoren, die das schichten-spezifische Einwachsen aussprossender Axone kontrollieren, trägt wesentlich zum Verständnis dieses Phänomens bei. Weiterhin wird die Aufklärung der zugrundeliegenden molekularen Faktoren für die spezifische Zielerkennung und deren Charakterisierung uns helfen, das Potential und die Limitation der Regeneration im ZNS besser zu verstehen und die Möglichkeit eröffnen, einmal verlorengegangene neuronale Verbindungen durch therapeutische Intervention wieder spezifisch aufzubauen. / In this study, the impact of membrane-associated factors on axonal outgrowth during development and following lesion was examined. We studied the maturation-dependent features of membrane-associated molecules in the hippocampus with the stripe assay for guidance activity and with the outgrowth assay for outgrowth-supporting activity. We could show that entorhinal axons discriminate between their proper target area, the hippocampus, and control regions which do not receive synaptic connections from the entorhinal cortex, and preferred to grow on hippocampal membranes. Further, we examined guidance preferences of entorhinal neurites on hippocampal membranes in different developmental stages. The choice behavior of entorhinal neurites for hippocampal membranes temporally correlates with the ingrowth of the perforant path into the hippocampus and with the stabilization of this brain area in vivo, and further indicate the transient presence of membrane-associated guidance cues in the hippocampus. One of the characteristics of maturational processes in the central nervous system is the developmentally regulated myelination of fiber tracts. Comparison of the stripe assay data with immunohistochemical analysis for MBP and MAG as representative myelin markers revealed a correlation between the changes in axonal choice behavior and increasing myelination. It is known that myelin itself is a strong axonal outgrowth inhibitor and that myelin can also induce growth cone collapse. In outgrowth assays, we could show that myelin has a strong outgrowth inhibitory influence on entorhinal axons which can be neutralized by the monoclonal antibody IN-1. However, in the stripe assay, myelin did not influence the choice behavior of outgrowing axons and this indicates that myelin does not govern information for directed growth. Furthermore, stripe assays were performed with membranes obtained from deafferented hippocampi at various lesion stages. In these experiments, we could show that outgrowth-promoting factors are present in the lesioned hippocampus. Moreover, data from the stripe assay revealed the timely restricted presence of membrane-bound guidance factors which are equally as attractive as neonatal hippocampal membranes. These experiments indicate the lesion-induced expression of outgrowth-promoting factors in the hippocampus, which correlates temporally with the sprouting reaction in vivo. It is suggested that the central nervous system of adult vertebrates is determined in its cellular condition and not capable of structural changes. This is most evident following lesion of adult brain, where the ability for regeneration is highly restricted in comparison to young, postnatal neural tissue. This restricted ability for regeneration in the adult brain is essentially determined by the presence of outgrowth-inhibitory myelin. However, a compensatory sprouting response exists in the adult hippocampus following lesion, which leads to a layer-specific replacement of lost synaptic contacts. The identification of these factors will lead to a deeper understanding of layer-specific axonal sprouting and synaptic replacement. Further, the identification and characterization of the underlying factors will help us to understand the potential and limitations of regeneration in the central nervous system.

Bacillus amyloliquefaciens FZB42 ist ein frei lebendes Bakterium, das Pflanzenwurzeln besiedelt und das Pflanzenwachstum durch viele verschiedene Wirkmechanismen anregt. In dieser Arbeit wurden die molekularen Grundlagen dieser positiven Wirkungen, die dieses „Pflanzenwachstum fördernde Rhizobakterium“ (PGPR) auf seine Wirte ausübt, untersucht. Um den gegenseitigen Austausch von B. amyloliquefaciens und seinen Wirtspflanzen zu entschlüsseln, wurden umfangreiche Proteomstudien durchgeführt. Es wurden Referenzkarten der extrazellulären und zytosolischen Proteinfraktionen erstellt. Die größte Anzahl an ausgeschiedenen Proteinen konnte während der stationären Phase beobachtet werden. Die identifizierten extrazellulären Proteine gehören verschiedenen Funktionsklassen an, wobei die prominentesten Klassen am Kohlenhydrat-Abbau und den Transport von Molekülen durch die Zellwand beteiligt sind. Die zytosolischen Extrakte von Kulturen, die in 1C-Medium bzw. Mineralmedium angezogen wurden, und in der zweidimensionalen Gelelektrophorese (2 DE) aufgetrennt wurden, ergaben 461 und 245 verschiedene Protein-Einträge. Die erstellten Referenz-Karten wurden anschließend verwendet, um Proteine und Prozesse, in an der Interaktion mit Pflanzen beteiligt sind, zu identifizieren. Dafür wurden die Bakterien Wurzelexudaten von Mais (Zea mays L.) ausgesetzt. Die Proteine aus zwei Stämmen, denen die globalen Transkriptionsregulatoren (Degu, AbrB) und vier Sigma-Faktoren (SigB, SigM, SigV, und SigX) fehlen, wurden ebenfalls untersucht, um ihre Beteiligung an den bakteriellen Reaktionen auf die Wurzelausscheidungen zu analysieren. Zusammenfassend ist dies die erste Studie, die umfangreiche Proteomdaten von Gram-positiven PGPR präsentiert, wobei gleichzeitig die Veränderung der Expression von extrazellulären und zytoplasmatischen Proteinen, nach Zugabe von Wurzelexudaten, ausgewertet wurde. / Bacillus amyloliquefaciens strain FZB42 is a free-living bacterium that competitively colonizes plant roots and stimulates plant growth by many different modes of action. The molecular basis of singular beneficial effects that this Plant Growth-Promoting Rhizobacteria (PGPR) exert on their hosts have been studied. To decipher the molecular cross-talk of B. amyloliquefaciens and its’ host plants as a whole system, an extensive proteomic approach was performed. Reference maps of the extracellular and cytosolic protein fractions were established. The highest number of secreted proteins was observed during stationary growth phase. Identified extracellular proteins belong to different functional classes, with the most prominent classes involved in carbohydrate degradation and transportation of molecules across the cell wall. Cytosolic extracts obtained from cultures grown in 1C and minimal media subjected to the 2 Dimensional Electrophoresis (2 DE), revealed 461 and 245 different protein entries, respectively. Created reference maps were subsequently used to identify proteins and processes involved in the interaction with plants, prior to exposure of bacteria to maize (Zea mays L.) root exudates. The proteomics of two strains lacking expression of genes coding for global transcriptional regulators (degU, abrB) and four sigma factors (sigB, sigM, sigV, and sigX) were also inves-tigated, in order to analyse their involvement in bacterial responses to root exudates. In summary, this is the first study presenting comprehensive proteomics of Gram-positive PGPR, evaluating at the same time changes in protein expression caused by addition of root exudates at the extracellular and cytosolic level.

Made available in DSpace on 2016-04-26T20:23:27Z (GMT). No. of bitstreams: 1 Maria Herminia Penteado Pacheco e Silva Moccia.pdf: 2132272 bytes, checksum: dd92a873e370f02c9e4e8afffe7c00a0 (MD5) Previous issue date: 2015-02-11 / This study, stemming from the provisions of the Brazilian Constitution, aims to analyze the activity of the State in fostering the private economy, so as to frame the latter as an expression of administrative activity, subject to the limits that govern this activity of the state. The wide range of activities fostering economic development results in a conceptual challenge in the attempt to materially determine its field of impact. Thus, among the instruments provided for in the Brazilian legal system, this study focused on the soft loans granted by the BNDES, in order to define the set of parameters to be applied to the exercise of this government activity. Actions to foster the economy, through soft loans in particular, has been treated as a result of the discretionary authority of the Administration, and considered an activity more connected to private and banking law, than to public law and its respective legal system. Although a greater flexibility in the choices made by the State is recognized, the public nature of this specific role of the State must be recognized. A role which should be motivated, transparent and controlled, in order to build boundaries and clarify the conditions for it to be carried out. This topic has not yet deserved in-depth studies, and has gained significance because of the enormous amounts of money that have been directed to this activity regardless of any guarantee of effectiveness of the spending. On the other hand, since 2009 the Bank started to receive resources from the Brazilian Treasury. These resources now outweigh the Bank s original sources of funding. This has led to an increase in public debt and huge distortions that undermine economic policy. In this study the role and the importance of public banks as funding instruments in developing economies is recognized. However, it is argued that the BNDES should be more parsimonious in granting loans at lower interest rates and guided by clear criteria, designed to effectively ensure economic and social development. Aware of the above mentioned challenges, and because of the different choices made by different Administrations in relation to each State s own economic reality, the ultimate goal of this study is to present instruments for establishing guiding parameters for this activity, and for enabling it to be effectively controlled / O presente trabalho, partindo dos comandos emanados da Constituição Federal, objetiva analisar a atividade estatal de fomento da economia privada, para enquadrá-la como expressão da atividade administrativa, sujeitando-a aos limites que informam esta atividade estatal. A ampla abrangência da atividade promocional, resulta em uma dificuldade conceitual para a delimitação material de seu campo de incidência. Assim, dentre os instrumentos previstos no ordenamento jurídico brasileiro, a tese focou nos empréstimos em condições favoráveis concedidos pelo BNDES, para definir os parâmetros aplicáveis ao seu exercício. O fomento, e este instrumento em específico, vem sendo tratado como resultado de escolha discricionária da Administração, considerada uma atividade mais ligada ao direito privado e bancário, do que ao direito público e seu regime jurídico. Embora se reconheça a maior flexibilidade nas escolhas estatais, deve-se ter em mira o caráter público desta função que deve ser motivada, transparente e controlada, com vistas a construir limites e explicitar os pressupostos para o seu exercício. Trata-se de um tema pouco estudado que ganhou relevância em razão das vultosas quantias que foram destinadas a esta atividade, sem garantia da eficácia do gasto empreendido. Por outro lado, desde 2009 o Banco passou a receber aportes do Tesouro que hoje superam suas fontes de custeio originárias, repercutindo no aumento da dívida pública e produzindo enormes distorções que prejudicam a política econômica. No presente trabalho são reconhecidos o papel e a importância dos Bancos Públicos como instrumentos de fomento, em economias em desenvolvimento. Ocorre que o BNDES precisa ser mais parcimonioso na concessão de empréstimos a juros favoráveis, e pautar-se por critérios claros, voltados a garantir, efetivamente, o desenvolvimento econômico e social. Ciente das dificuldades acima aludidas, e em razão das diferentes escolhas das atuações administrativas em relação com a realidade econômica própria de cada Estado, o objetivo final deste trabalho é apresentar instrumentos que permitam a formulação de parâmetros norteadores desta atividade, auxiliando o seu controle

The inoculation with Azospirillum brasilense is one of the main strategies to supplement the inorganic inputs of nitrogen (N) and to increase the root development in maize. However, the beneficial inoculation effects are not always reached, which, in part, is due to genotypic variation in the plant host, resulting in different degrees of outcome. In this context, we aimed to study the genetic control and genomic prediction of maize traits related to the responsiveness to A. brasilense inoculation. For this, 118 maize hybrids were conducted under N stress and N stress plus A. brasilense treatments in controlled conditions over 2016 and 2017 seasons. We evaluated root and shoot traits and performed diallel analyses, association mapping, and genomic prediction methods considering 59,215 Single-Nucleotide Polymorphism (SNP) markers. Our results revealed a quantitative inheritance of the partnership-related maize traits, with both additive and non-additive genetic effects involved in the genetic control. Furthermore, several candidate genes were identified for the maize-A. brasilense association, especially with heterozygous (dis)advantage effects. In general, the prediction accuracies were higher mostly for the inoculated treatment compared to the non-inoculated. Finally, our findings enable a deeper understanding towards the genetic basis of the maize responsiveness to A. brasilense and may support plant breeders to establish selection strategies aiming the development of superior genotypes for this association. / A inoculação com Azospirillum brasilense é uma das principais estratégias para suplementar os insumos inorgânicos de nitrogênio (N) e aumentar o desenvolvimento radicular do milho. No entanto, os efeitos benéficos da inoculação nem sempre são alcançados, o que, em parte, é devido à variação genotípica da planta hospedeira, que ocasiona diferentes graus de resultados. Neste contexto, nosso objetivo foi estudar o controle genético e a predição genômica de caracteres de milho relacionados à responsividade para a inoculação com A brasilense. Para isso, 118 híbridos de milho foram conduzidos sob estresse de N e estresse de N mais A brasilense em condições controladas nos anos de 2016 e 2017. Nós avaliamos características de raiz e parte aérea e realizamos análises dialélicas, mapeamento associativo e métodos de predição genômica considerando 59.215 marcadores Single-Nucleotide Polymorphism (SNP). Nossos resultados revelaram uma herança quantitativa das características do milho relacionadas à essa parceria, com efeitos genéticos aditivos e não-aditivos envolvidos no controle genético. Além disso, vários genes candidatos foram encontrados para a associação milho-A brasilense, especialmente com efeitos de (des)vantagens de heterozigotos. Em geral, as acurácias de predição foram mais maiores principalmente para o tratamento inoculado em comparação ao não inoculado. Finalmente, nossos resultados possibilitam uma compreensão mais aprofundada das bases genéticas da responsividade do milho à A. brasilense e podem auxiliar os melhoristas de plantas a estabelecerem estratégias de seleção visando o desenvolvimento de genótipos superiores para essa associação.