In this thesis I do a comparative study between the Brazilian Portuguese Grammar (henceforth BP) and European Portuguese Grammar (henceforth EP) concerning to the word order, more specifically, with respect to the subject order in relation to the verb in finite declarative phrases. One defends that Verb-Subject order (VS) has been lost in the first language in (in)transitive contexts due a parametric change in the marking of Null Subject Parameter caused by weakness of AGR, what implies to consider that BP is losing the characterization of a prototypical null subject language as the EP for become a non-prodrop subject language as English. In this order, in inaccusative contexts, the visible morphologically agreement between a verbal inflection and the post-verbal DP is generally not observed and this DP can also be definite or indefinite, unlike what is proposed by Belletti s approach. Futhermore, researches have assumed that the subject position with respect to the verb comes from the fact of the syntax be conditioned by discoursive constraints. Therefore, with respect to these aspects, the goals of this research are: a) to develop an analysis about VS order in both grammars, considering the implications of the AGR richness for the position of the subjects ; b) to characterize this richness from the rediscussion of some theoretical proposals; c) to show counter-evidences to the proposal of that the word order codifies the informational structure; d) to explain the possibility of the non-visible morphologically agreement between the post-verbal DP and the verbal inflection in inaccusative contexts and its implications for the type of mechanism concerned to the nominative Case assignment and e) to argue against the universality of the Definiteness Efect on the post-verbal DP. To develope this study, the analysis bases itself on Pricinciples and Parameters, and the Distributed Morphology frameworks. The sentence structures are results from introspection data. During this research, it was possible to conclude that rich AGR that licences and identifies referential null subjects in null subject languages has no correlation with rich AGR that causes the verb movement in the syntax, and that preverbal subjects occupy an A-position in BP and in EP, unlike what is generally proposed. Futhermore, the different behaviour of VS order observed among these languages receives explanatory support from the type of syntactic output generated by each grammar in particular but not from discoursive constraints. With respect to the order in which there is no visibility of morphological agreement between the verbalinflection and the post-verbal DP in inaccusative contexts, I argue that it is a pseudo- VS order, in which Definiteness Efect does not act on. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Nesta tese realizo um estudo comparativo entre a gramática do português brasileiro (doravante PB) e a do português europeu (doravante PE) no que diz respeito à ordem dos constituintes, mais especificamente, do sujeito em relação ao verbo em frases declarativas finitas. Tem-se defendido que a ordem Verbo-Sujeito (VS) tem sido perdida na primeira língua em contextos (in)transitivos em virtude de uma mudança paramétrica na marcação do Parâmetro do Sujeito Nulo ocasionada pelo enfraquecimento de AGR, o que implica considerar que o PB está deixando de ser uma língua de sujeito nulo prototípica como o PE para tornar-se uma língua de sujeito não-nulo como o inglês. Nessa ordem, em contextos inacusativos, a concordância morfologicamente visível entre a flexão verbal e o DP pós-verbal, em geral, não é observada e, ainda, esse DP pode ser definido ou indefinido, ao contrário do que é proposto pela análise belletiana. Ademais, estudos têm assumido que a posição do sujeito em relação ao verbo decorre do fato de a sintaxe estar a serviço de requerimentos de ordem discursiva. Portanto, face a esses aspectos, os objetivos desta pesquisa são: a) desenvolver uma análise sobre a ordem VS em ambas as gramáticas, levando em conta as implicações da riqueza de AGR para a posição dos sujeitos; b) caracterizar essa riqueza , a partir da rediscussão de algumas propostas teóricas; c) apresentar contra-evidências à proposta de que a ordem dos constituintes codifica a estrutura informacional; d) explicar a possibilidade de concordância morfologicamente não-vísível entre o DP pós-verbal e a flexão verbal em contextos inacusativos e suas implicações para o tipo de mecanismo relacionado à atribuição de Caso nominativo e e) argumentar contra a universalidade do Efeito de Definitude sobre o DP pós-verbal. Para a realização desse estudo, a análise fundamenta-se no arcabouço teórico do Modelo de Princípios e Parâmetros e da Morfologia Distribuída, sendo as estruturas frasais resultados de dados de introspecção. No decorrer desta pesquisa, é plausível concluir que AGR rico que licencia e idendifica sujeitos nulos referenciais em línguas de sujeito nulo não tem a ver com AGR rico que motiva o movimento do verbo na sintaxe, e que sujeitos pré-verbais ocupam uma posição-A no PB e no PE, ao contrário do que é comumente proposto na literatura. De mais a mais, o comportamento diferenciado da ordem VS atestado entre essas línguas ganha suporte explicativo no tipo de output sintático gerado por cada gramática em particular e não em requerimentos de ordem discursiva. No que concerne à ordem em que não há visibilidade da concordância morfológica entre a flexão-verbal e o DP pós-verbal em contextos inacusativos, argumento que seja uma pseudo-ordem VS , não sendo atuante sobre esse DP o Efeito de Definitude.

Background: Innovation and collaboration are deemed popular terms that are widely used and agreed on. It is implied that innovation without collaboration seems to be unlikely (Deichmann et al., 2017, Haanæs et al., 2018, Innov8rs, 2018). However, the meaning behind innovations of disruptive, incremental, or radical character remains vague and presumably differs between practitioners, just as it differs among scholars. Also, the literature only implies how collaboration can be used effectively for innovation, whilst focussing more on what forms of collaborations are existing.   Research questions: How is innovation understood in the CIC? How does the CIC work together in order to realise cross-industry collaborations for innovation?   Purpose: This study aims at investigating the variety of innovation understanding and how it is applied in collaborations, based on the views of practitioners from different companies that are part of the Cross Industry Club.   Method: This research is designed as an exploratory case study and follows a qualitative strategy with abductive reasoning. Data is collected through nine semi-structured interviews with representatives from five different companies.   Conclusion: The study revealed that practitioners have other aspects in mind which define their understanding of innovation compared to scholars. By that, the usage of innovation terminology is rather arbitrary in practice. The influence of innovation frameworks on the understanding of innovation and the collaborative work have been revealed. Innovation in a cross-industry collaboration, in its purpose and characteristics, is similar to the work of communities of practice. Challenges that appear during the emergence of a collaboration like the CIC, and values that affect the initial work towards cross-industry projects, have been identified.

Orientador: José Francisco Comenalli Marques Júnior / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-21T08:08:20Z (GMT). No. of bitstreams: 1 Moraes_MaraCabral_M.pdf: 399240 bytes, checksum: bb189c0420b266a0b620156bd10cd435 (MD5) Previous issue date: 2012 / Resumo: INTRODUÇÃO: A ILD vem sendo utilizada como opção terapêutica para recaída de neoplasias hematológicas após transplante alogênico. Contudo, este procedimento não é isento de riscos e sua indicação deve ser criteriosa. Neste trabalho avaliamos a ILD quanto à eficácia da coleta; efeitos adversos para os doadores e pacientes e efeitos quanto à remissão da doença, redução de recaída e mortalidade. MÉTODOS: Foi realizado estudo retrospectivo dos pacientes (n=40) que receberam ILD, no Hemocentro da UNICAMP, de março 1995 a janeiro 2008, e dos respectivos doadores (n=40). As características dos pacientes, complicações e sobrevida foram comparadas a um grupo de pacientes (n=264) que realizou transplante alogênico e não recebeu ILD. RESULTADOS: As 54 coletas de linfócitos analisadas atingiram o alvo estabelecido de '10 POT. 7' CD3+/Kg receptor, sem nenhuma reação grave para os doadores. Não observamos diferença estatística na mortalidade (p= 0,85) ou incidência de DECHa (p=0,231) entre os grupos de pacientes. No entanto, as causas de óbito foram diferentes (p<0,001), com maior mortalidade por doença no grupo em estudo e por outras causas no grupo comparativo. A incidência de DECHc e remissão completa foram maiores no grupo comparativo (p<0,001), enquanto o tempo para o óbito foi maior no grupo em estudo (p=0,009). CONCLUSÕES: A coleta de linfócitos foi eficaz e segura para o doador. Para os pacientes a DECH se sobressaiu como efeito adverso, sem aumento após a ILD. Não foi observado aumento na remissão da doença ou queda na mortalidade após a ILD, contudo, o tempo para o óbito foi maior. Outros fatores e a heterogeneidade das amostras podem ter contribuído para os resultados encontrados / Abstract: BACKGROUND: Donor lymphocyte infusion (DLI) has been used as a therapeutic option for hematological recurrences after allogeneic transplantation. However, this procedure is not devoid of risks and its indication must be precise. In this article we assess DLI regarding collection efficacy; adverse effects in donors and patients and its impact on disease remission, recurrence induction and mortality. METHODS: A retrospective study of patients (n=40) who received DLI and their respective donors was performed at Hemocentro, UNICAMP, between March 1995 and January 2008. Patient characteristics, complications and survival were compared to a comparative group (n=264) consisting of patients who underwent allogeneic transplantation, but did not receive DLI. RESULTS: All the 54 lymphocyte collection procedures reviewed reached the set target of '10 POT. 7' CD3+/Kg receptor, with no serious adverse reactions to the donors. We did not observe statistical difference in mortality (p=0.85) nor in the incidence of acute GVHD (p=0.231) between the two patient groups. However, causes of death were statistically different (p<0.001), with a higher disease-related mortality rate in the study group and for other causes in the comparative group. Chronic GVHD incidence and complete remission were higher in the comparative group (p<0.001), while the overall survival was longer in the study group (p=0.008). CONCLUSIONS: Lymphocyte collection was efficient and safe to the donor. As for patients, GHVD was the most important side effect but did not have a higher incidence after DLI. We did not observe an increase in disease remission or decrease in mortality rate after DLI; however, overall survival was longer. Other factors and the sample heterogeneity might have contributed to the results observed / Mestrado / Clinica Medica / Mestra em Clínica Médica

Ces travaux abordent la question de l'impact du pouvoir et du contexte privé ou public sur la sensibilité à l’influence normative. Nous défendons la thèse selon laquelle le contexte régule les effets du pouvoir sur la sensibilité à l’influence normative. Dans une première partie, nous avons testé l'hypothèse selon laquelle la possession de pouvoir, par la désinhibition qu’il suscite, rendrait les individus moins sensibles à la norme. Nous présentons deux études qui montrent que la possession de pouvoir occasionne des comportements moins conformes aux normes ainsi qu’une présentation de soi socialement moins désirable. Dans une seconde partie, nous testons l’hypothèse selon laquelle l’activation d’un pouvoir élevé entrainerait une plus forte désinhibition des conduites en privé et une plus forte inhibition des conduites en public que l’activation d’un pouvoir faible. Deux études ont été réalisées et tendent à soutenir cette idée. Une dernière recherche a été menée en vue d’examiner le rôle de l’activation contextuelle de la responsabilité sociale sur la relation entre le pouvoir et la sensibilité à l’influence normative. Toutefois, il semble que l’activation de la responsabilité sociale ne médiatise pas les effets modérateurs du contexte sur la normativité des jugements effectués. Dans l'ensemble, ces travaux contribuent à éclaircir les effets paradoxaux du pouvoir qui ont été observés dans la littérature, en proposant un modèle permettant de comprendre comment le contexte génère des processus différents en fonction du pouvoir, quand la désinhibition et l’inhibition sont susceptibles de se produire, et quelles en sont les conséquences sur les conduites individuelles. / This work investigates the impact of social power and private or public context on normative influence. We defend the thesis according to which the context (private vs. public) regulates the effects of power on normative influence. In a first series of research, we tested the hypothesis that the possession of power, by generating disinhibition, would make individuals less sensitive to social norms. Two studies are presented and show that the possession of power causes less normative behaviors as well as less socially desirable self-presentations. In a second series of research, we argue that high power primed individuals would exhibit greater disinhibition in private and greater inhibition in public than low power primed individuals. Two studies were conducted and tend to support this idea. A last study examined social responsibility as a potential moderator of the effects of social power and context on normative influence. However, it seems that social responsibility does not mediates the inhibition and disinhibition patterns we observe. Overall, these studies help to clarify the paradoxical effects of power underlined in the literature, by proposing a model which allows to understand how the context generates different processes depending on social power, when disinhibition and inhibition are likely to occur, and what are the consequences for individual behavior.

TITLE: Contemporary art at school - post internet age AUTHOR: Nikola Tora Dvořáková DEPARTMENT: Department of art education SUPERVISOR: Ph.Dr. Leonora Kitzbergerová, PhD. ABSTRACT: The thesis - Contemporary art at school - post internet age is about current arts occasions, concretely Post internet art. I try to state and define this kind of art and place post internet art to context with art education afterwards. Specific art pieces and their similarities in related zones of art are reflected in didactic transformations. The research is focused on difficulties, which can come into the art education lessons. It is realized with different methods, on different kind of school. In concluision my original art piece is bringing more comprehensive knowledge about post internet art. KEYWORDS: Contemporary art, post internet art, interpreter vs. consumer, new media

Les ribozymes sont des ARN catalytiques fréquemment exploités pour le développement d’outils biochimiques et d’agents thérapeutiques. Ils sont particulièrement intéressants pour effectuer l’inactivation de gènes, en permettant la dégradation d’ARNm ou d’ARN viraux associés à des maladies. Les ribozymes les plus utilisés en ce moment pour le développement d’agents thérapeutiques sont les ribozymes hammerhead et hairpin, qui permettent la reconnaissance spécifique d’ARN simple brin par la formation de structures secondaires stables. In vivo, la majorité des ARN adoptent des structures secondaires et tertiaires complexes et les régions simples brins sont parfois difficiles d’accès. Il serait intéressant de pouvoir cibler des ARN repliés et un motif d’ARN intéressant à cibler est la tige-boucle d’ARN qui peut être importante dans le repliement global des ARN et pour accomplir des fonctions biologiques. Le ribozyme VS de Neurospora fait la reconnaissance de son substrat replié en tigeboucle de façon spécifique par une interaction kissing-loop, mais il n’a jamais été exploité pour faire la reconnaissance d’un ARN cible très différent de son substrat naturel. Le but des travaux présentés dans cette thèse est de déterminer si le ribozyme VS possède l’adaptabilité nécessaire pour l’ingénierie de ribozymes qui clivent des ARN cibles différents du substrat naturel. Dans le cadre de cette thèse, le ribozyme VS a été modifié pour l’adapter à différents substrats et des études de cinétiques ont été réalisées pour évaluer l’impact de ces modifications sur l’activité de clivage du ribozyme. Dans un premier temps, le ribozyme a été modifié pour faire la reconnaissance et le clivage de substrats possédant différentes longueurs de tiges Ib. Le ribozyme a été adapté avec succès à ces substrats de différentes longueurs de tige Ib, avec une activité qui est similaire à celle du ribozyme avec un substrat sans modification. Dans un deuxième temps, c’est l’interaction kissing-loop I/V du ribozyme qui a été substituée de façon rationnelle, dans le but de savoir si un ribozyme VS mutant peut reconnaitre et cliver un substrat ayant une boucle différente de celle de son substrat naturel. L’interaction kissing-loop I/V a été substituée pour les interactions kissing-loop TAR/TAR* de l’ARN du VIH-1 et L22/L88 de l’ARN 23S de Deinococcus radiodurans. La réaction de iii clivage des ribozymes comportant ces nouvelles interactions kissing-loop est toujours observée, mais avec une activité diminuée. Finalement, la sélection in vitro (SELEX) de ribozymes a été effectuée pour permettre un clivage plus efficace d’un substrat mutant avec une nouvelle boucle. Le SELEX a permis la sélection d’un ribozyme qui clive un substrat avec une boucle terminale mutée pour celle de l’ARN TAR du VIH-1 et cela avec une activité de clivage très efficace. L’ensemble de ces études démontre que le ribozyme VS peut être modifié de diverses façons pour la reconnaissance spécifique de différents substrats, tout en conservant une bonne activité de clivage. Ces résultats montrent le grand potentiel d’ingénierie du ribozyme VS et sont prometteurs pour la poursuite d’études d’ingénierie du ribozyme VS, en vue du clivage d’ARN cibles repliés en tige-boucle complètement différents du substrat naturel du ribozyme VS. / Ribozymes are catalytic RNAs frequently exploited for the development of biochemistry tools and therapeutic agents. They are particularly interesting in gene inactivation strategies for the degradation of mRNA and viral RNA genome. Currently, the most common ribozymes used in the development of therapeutic agents are the hammerhead and hairpin ribozymes, which can specifically recognize and cleave target single-stranded RNAs through the formation of stable secondary structures. In vivo, single-stranded RNAs can be difficult to target because most RNA adopt complex secondary and tertiary structures. It could be worthwhile to target folded RNA motifs, and an interesting target is the stem-loop structure because stem-loops are important for the overall folding of RNA molecules and they can perform important biological functions. The Neurospora VS ribozyme recognizes its stem-loop folded substrate via a specific kissing-loop interaction, but it has never been exploited for the recognition of target RNA very different from its natural substrate. The goal of the work presented in this thesis is to determine whether the VS ribozyme possesses the necessary adaptability for engineering ribozymes that target RNAs different from its natural substrate. For this thesis, the VS ribozyme was adapted for the recognition of different substrates and kinetic studies were performed to evaluate the effect of these modifications on the cleavage activity. In a first study, the VS ribozyme was modified to recognize and cleave substrates with different stem Ib lengths. The VS ribozyme was successfully adapted to theses substrates of different lengths with a cleavage activity similar to the unmodified ribozyme and substrate. In a second study, the I/V kissing-loop interaction was substituted by rational design, in order to establish if the VS ribozyme variants could recognize and cleave a substrate with a different loop than its natural substrate. The I/V kissing-loop interaction was substituted for the HIV-1 TAR/TAR* and the Deinococcus radiodurans RNA large rRNA subunit L22/L88 kissing-loop interactions. The cleavage reaction was observed for the ribozymes with these new interactions, but with reduced activity. Finally, in vitro selection (SELEX) was used to enable more efficient cleavage of a mutant substrate with a new loop. SELEX experiments enabled v the selection of ribozyme variants that cleave a substrate with a terminal loop mutated to that of the HIV-1 TAR RNA with a very efficient cleavage activity. All of the studies presented in this thesis show that the VS ribozyme can be modified in various ways for the specific recognition of different substrates, while maintaining efficient cleavage activity. These results demonstrate the great potential of VS ribozyme engineering and are very promising for further engineering studies of VS-derived ribozymes for the cleavage of stem-loop folded target RNA completely different from its natural VS ribozyme substrate.

Ce document constitue l'aboutissement de trois années de thèse. Après avoir introduit et dégagé la problématique générale se rapportant à mon sujet de thèse, à savoir « comment utiliser les données de provenance pour favoriser un Cloud de confiance ? », je présente une description des concepts, modèles et langages se rapportant à ma thèse et l'état de l'art qui peut répondre en partie à cette problématique. En second lieu, je présente la solution basée sur la provenance que j'apporte aux contrôles d'accès, dans les systèmes distribués comme le Cloud : PBAC². Elle repose sur un système combinant à la fois des modèles de provenance (PROV-DM) et de contrôles d'accès (règles génériques de type RBAC avec des politiques d'embrigadement ou de réglementation). Ce système utilise un moteur d'exécution central appelé le médiateur pour renforcer la sécurité et favoriser la confiance dans le Cloud, via la vérification de règles sur une partie du graphe de provenance rétrospective qu'il a reçue. Par ailleurs, je décris l'étude que j'ai faite de trois extensions de PBAC² : (1) l'intégration de l'ontologie PROV-O et ses avantages et inconvénients quant à la taille du (sous-)graphe de provenance reçu par le médiateur ; (2) la construction de l'adaptation de PBAC² avec l'approche de sécurité qu'est la réglementation; (3) la traduction des règles PBAC² en contraintes PROV-CONSTRAINTS. De plus, PBAC² est appliqué sur un exemple réaliste propre au secteur médical. Un prototype de PBAC² et une démonstration sur des exemples concrets avec une machine locale et un système de Cloud réel illustrent la portée de ce travail. En conclusion de la thèse, je propose quatre perspectives de ce travail. / This document is the culmination of three years of thesis. Having introduced and cleared the general issue related to my thesis subject, i.e. « how to use provenance data to enforce trust in the Cloud? », I present a description of the concepts, models and languages related to my thesis and the state of the art that can partially address this issue. Secondly, I present the solution based on provenance that I bring to access controls, in distributed systems such as the Cloud: PBAC². It is based on a system combining both provenance models (PROV-DM) and access controls (generic rules of RBAC type with regimentation and regulation policies). This system uses a central execution engine denoted the mediator to enforce security and foster trust in the Cloud, via rule checking over a part of the retrospective provenance graph it received. Furthermore, I describe the study I made of three PBAC² extensions: (1) the integration of the PROV-O ontology and its pros and cons regarding the size of the (sub)graph received by the mediator; (2) the construction of the PBAC² adaptation with the regulation security approach; (3) the translation of PBAC² rules into PROV CONSTRAINTS constraints. Moreover, PBAC² is applied to a realistic example that belongs to the healthcare sector. A PBAC² prototype and a demonstration on some practical examples with a local machine and a real Cloud system illustrate the scope of this work. In conclusion of the thesis, I propose four perspectives of this work.

No experimento I, objetivou-se avaliar os efeitos de aditivos microbianos e tempo de armazenamento na qualidade de silagens de milho. A cultura do milho foi ensilada sem aditivos (Controle) ou com inoculantes contendo bactérias láticas homofermentativas (Lactobacillus plantarum + Enterococcus faecium + Pediococcus acidilactici) + enzimas celulolíticas e hemiceluloliticas ou heterofermentativas (Lactobacillus buchneri) aplicados na dose 1 × 105 ufc/g. As silagens foram armazenadas em silos experimentais durante 3, 7, 15, 30, 60, 210, 390, 480 ou 570 dias. Foram avaliados: composição químico-bromatológica, produtos de fermentação, perfil microbiológico, perdas fermentativas, estabilidade aeróbia e degradabilidade ruminal in situ. Os inoculantes não afetaram a maior parte das variáveis estudadas. Entretanto, L. buchneri aumentou a concentração de ácido acético das silagens e diminuiu a deterioração aeróbia, confirmada pelo menor acúmulo térmico durante o ensaio de estabilidade aeróbia. O teor de carboidratos solúveis diminuiu ao longo do tempo de armazenamento, reflexo do metabolismo dos açúcares em produtos de fermentação. As concentrações da prolamina como esperado diminuíram e as concentrações de nitrogênio amoniacal e proteína solúvel aumentaram com os tempos de armazenagem prolongados, reflexo da ocorrência de proteólise na silagem de milho. O pH da silagem diminuiu rapidamente nos primeiros sete dias de armazenamentos mantendo-se estável para os demais tempos de estocagem, diferente do ocorrido para a concentração de ácido acético, que aumentou com o tempo de armazenamento. Verificou-se diminuição na contagem de bactérias láticas e leveduras ao longo do armazenamento. A produção de gás e a perda de matéria seca aumentaram com o tempo de armazenamento. O tempo de armazenamento aumentou todas as variáveis de EA. Os ganhos mais significativos para a EA se deram até aproximadamente 60 dias de armazenamento. A degradação ruminal de amido e, consequentemente de MS, foi incrementada ao longo do armazenamento. No experimento II, objetivou-se avaliar o desempenho de vacas leiteiras em função da estratégia de descarregamento da silagem de milho em silo do tipo trincheira: silagem de milho oriunda da metade superior do silo (topo) ou silagem de milho oriunda da metade inferior do silo (base). Foram utilizadas 24 vacas alocadas em 12 blocos casualisados, com arranjo de reversão simples com períodos de 21 dias. Os animais foram alojados em confinamento tie-stall. As dietas foram iso-protéicas (16,5%) e iso-amiláceas (17,0%), com 60% de silagem de milho (% MS). O consumo de matéria seca, produção e composição do leite foram determinados entre os dias 15 e 21 de cada período experimental. Apesar da silagem oriunda da base do silo levar à maior digestibilidade da dieta e menor concentração de nitrogênio ureico do leite (8,95 e 11,35 mg/dL) não houve efeito da dieta no consumo de matéria seca nem na produção de leite. Sob condições ótimas de manejo, a estratégia de descarregamento da silagem de milho não afeta o desempenho de vacas leiteiras. / In the experiment I, the aim was to evaluate the effects of microbial additives and length of storage on the quality of corn silage. Whole-corn plants were ensiled without or with inoculants containing homofermentative (Lactobacillus plantarum + Enterococcus faecium + Pediococcus acidilactici + cellulolytic and hemicellulolytic enzymes) or heterofermentative lactic acid bacteria (Lactobacillus buchneri) applied at 1 × 105 cfu/g. Treated forages were packed and stored in experimental silos for 3, 7, 15, 30, 60, 210, 390, 480, and 570 days. Samples were evaluated for chemical composition, fermentation end-products, microbial counts, fermentation losses, aerobic stability, and ruminal degradability in situ. Inoculants did not affect most of the variables studied. However, L. buchneri increased acetic acid concentration and decreased aerobic deterioration of silages, as indicated by the lower heat accumulation during the exposure to air. Soluble carbohydrates decreased across the storage period, reflecting the conversion of soluble sugars to fermentation end-products. Concentrations of prolamin decreased, whereas ammonia and soluble protein concentrations increased over the storage period, indicating the proteolysis. The silage pH declined rapidly in the first seven days of storage and remained stable for the remaining storage period, unlike for acetic acid concentration that increased with storage period. On the other hand, counts of lactic acid bacteria and yeasts decreased during the storage. Gas production and dry matter loss increased with the length of storage. The storage period increased all variables stability aerobic. Most of significant improvements in stability aerobic were observed during the first 60 days of storage. Ruminal degradability of starch and, consequently, dry matter increased along the storage. In the experiment II, the aim was to evaluate the influence of strategy of silage unload on the performance of dairy cows. Corn silage from a bunker silo was separated at unloading as silage from the upper half of the silo (top) or from the lower half of the silo (bottom) and used to compose total mixed rations fed to 24 lactating cows allocated in 12 randomized blocks, arranged in a cross-over design with 21 periods. Cows were housed in a tie-stall barn. All diets contained 60% of corn silage and were iso-nitrogen (16.5% CP) and iso-starch (17.0% of starch). Dry matter intake, milk yield and composition were determined from day 15 to 21 in each period. Although silage from bottom led to higher total tract DM digestibility and lower milk urea nitrogen concentration (8.95 vs. 11.35 mg/dL), most of evaluated variables were not affected by treatments. Under optimal silo management, the strategy used to unload corn silage does not affect the performance of dairy cows.

De par l'omniprésence des rayonnements ionisants, l’évaluation de leur impact sur les écosystèmes est devenue une préoccupation environnementale majeure. Cependant, l’évaluation des risques environnementaux liés aux expositions chroniques souffre d’un manque de connaissances, d’autant que l’extrapolation des données acquises après exposition aiguë (plus nombreuses) ne semble pas adaptée pour la prédiction des effets des expositions chroniques. Pour exemple, les effets sur les paramètres individuels i.e. la reproduction, diffèrent entre ces deux modes d’expositions, suggérant que les mécanismes moléculaires sous-jacents diffèrent également. Il est donc nécessaire de réaliser des études au niveau individuel et subcellulaire afin de mieux comprendre les différences de radiotoxicité observées entre les deux modes d’irradiation. Les protéines sont les molécules fonctionnelles dans les organismes, elles peuvent être les cibles de dommages oxydatifs (i.e carbonylation), et sont susceptibles d’être des marqueurs pertinents et sensibles de l’exposition aux rayonnements ionisants. Ainsi, l’objectif de ce projet de thèse était d’améliorer la compréhension des mécanismes moléculaires de radiotoxicité (aigu vs chronique) en étudiant particulièrement la contribution du protéome chez un organisme modèle, le nématode Caenorhabditis elegans. Pour ce faire, l’étude des effets d’irradiation gamma aiguë et chronique sur une large gamme de doses (0,5 - 200Gy, dont 4 doses communes aux deux modes d’exposition) a été opérée au niveau individuel, et en particulier sur la reproduction, paramètre susceptible d’influencer directement la dynamique des populations. En complément, la modulation de l'expression des protéines mais aussi leurs dommages (i.e carbonylation) et leur dégradation par le protéasome ont été évalués. Les résultats ont montré que l’irradiation aiguë induit un effet sur le succès d’éclosion et sur la ponte totale dès 30Gy alors que seule la ponte totale est impactée par l’irradiation chronique à partir de 3,3Gy. A l’échelle moléculaire, les niveaux de protéines carbonylées sont très peu modulés après exposition chronique ou aiguë aux rayonnements ionisants. Le protéasome semble être impliqué dans la dégradation des protéines carbonylées après irradiation chronique. En revanche, après irradiation aiguë, celui-ci semble dépassé, suggérant une possible implication d’autres mécanismes de défense (autophagie). Les profils d’expression des protéines, et notamment de protéines impliquées dans l’apoptose, la réparation des dommages à l’ADN, la réplication et la reproduction sont différents après irradiation aiguë et chronique. Ainsi, les protéines nécessaires au développement embryonnaire sont réprimées après irradiation aiguë dès 0,5 Gy alors que celles impliquées dans le développement de la lignée germinale sont surexprimées. Ces dernières sont réprimées après irradiation chronique dès 0,5 Gy. Ces résultats, suggèrent que les mécanismes de radiotoxicité entre les expositions aiguës et chroniques sont bien distincts, et que les effets de l’irradiation aiguë pourraient être dus à une perturbation de l’embryogénèse (via l’accumulation de dommages génotoxiques). A l’inverse, l’irradiation chronique induirait un effet sur la gamétogénèse se traduisant par une baisse de la ponte totale sans affecter l’embryogénèse. Ce projet de recherche nous a permis d’apporter des connaissances sur les cascades d’évènements moléculaires suite à différentes conditions d'irradiation gamma et illustre l’intérêt d’utiliser une approche intégrée pour mieux prédire et comprendre les effets observés sur les grandes fonctions biologiques. De plus ces travaux ont permis de caractériser des marqueurs protéiques d’exposition plus sensibles que les marqueurs individuels puisque l’activité du protéasome et l’expression des protéines est modulée dès 0,5Gy. In fine cet ensemble de données contribuera à améliorer l’évaluation des risques pour l’environnement. / Because of the ubiquitous nature of ionizing radiation, the risk assessment on ecosystems has become a major environmental concern. However, the environmental risk assessment of chronic exposures suffers from a lack of knowledge, especially because the extrapolation of data acquired after acute exposure in order to predict the effects of chronic exposures is not always relevant. Indeed, the effects on the individual parameters, i.e reproduction, differ between these two irradiation modes, suggesting that underlying mechanisms are also different. It is therefore necessary to carry out studies at the individual and at the subcellular level in order to better understand molecular mechanisms governing these differences in the observed effects. Proteins are the functional molecules in organisms, they can be the targets of oxidative damage (i.e carbonylation), and are likely to be relevant and sensitive markers of exposure to ionizing radiation. Thus, the objective of this research project was to improve the understanding of molecular mechanisms of radiotoxicity (acute vs chronic), particularly by studying the proteome contribution, on the biological model Caenorhabditis elegans. The study of the acute and chronic gamma irradiation effects, on a large dose range (between 0.5 and 200Gy, including 4 common doses to both irradiation modes), was performed at the individual level with the reproduction as endpoint, a parameter likely to directly influence the dynamic of populations. In addition, the modulation of protein expression but also their damage (i.e. carbonylation) and their degradation by the proteasome were evaluated. The results showed that acute irradiation induced an effect on hatching success and on total spawning from 30 Gy whereas only total spawning was impacted after chronic irradiation from 3.3 Gy. At the molecular level, the global level of carbonylated proteins was not so modified after chronic or acute exposure to ionizing radiation. The proteasome appears to be involved in the degradation of carbonylated proteins after chronic irradiation whereas after acute irradiation, it seems overtaken, suggesting a possible involvement of other defense mechanisms (autophagy). The protein expression, and particularly proteins involved in apoptosis, DNA repair, replication and reproduction, is differentially modulated after acute and chronic exposure. Thus, the proteins involved in embryonic development are repressed after acute irradiation as soon as 0.5 Gy whereas those involved in the germline development are overexpressed. These results suggest that the radiotoxicity mechanisms between acute and chronic exposures are quite different and that the effects of acute irradiation may be due to an embryogenesis disturbance (via the accumulation of genotoxic damage). Conversely to acute, chronic irradiation induces an effect on gametogenesis, resulting in a decrease of the total spawning without impacting embryogenesis. This research project allowed us to provide knowledge on the molecular cascade events following different gamma irradiation conditions and highlights the need of using an integrated approach to better predict and understand the observed effects on major biological functions. Moreover, this work allowed characterizing more sensitive markers of exposure than the individual ones as the proteasome activity and the protein expression is modulated from 0.5Gy. Ultimately this dataset would help to improve the environmental risk assessment.

The relative importance of local and regional factors to community build-up is a core issue in contemporary ecology. Studies based on research of newly created aquatic habitats may improve understanding of these processes. We monitored zooplankton colonization rates and community assemblage in 20 newly dug and isolated temporary pools in Kokořínsko Protected Landscape Area. The region is characterised by deep stream valleys, extensive land use, low human influence, low waterfowl migration activity and practically absence of large water bodies. After the first hyproperiod all temporary pools were rebuilt to a permanent form. In contrast to expectations, in both cases we observed immediate colonization of newly created habitats by two main groups of zooplankton (crustaceans and rotifers), but only rotifers (Rotifera) were capable of fast successful establishment of viable populations. Cladocerans (Crustacea: Cladocera) and copepods (Crustacea: Copepoda) were observed rarely and usually in low abundances (< 1 ind-l ). 10 months after the filling of permanent pools one ostracod species (Crustacea: Ostracoda), Notodromas monacha, was recorded as well. During the 8 months of temporary pools existence the presence of 1 cladoceran and 1 cyclopoid copepod species and 12 rotifer taxa was recorded, icluding...