Limitierend für pharmakologische Therapien ist oft die Unfähigkeit des Wirkstoffes, biologische Membranen zu überwinden, weswegen häufig Transportmoleküle wie z.B. zellpenetrierende Peptide (CPPs, cell penetrating peptides) benutzt werden. Von den über 100 beschriebenen CPPs wurde bisher nur eine kleine Anzahl systematisch verglichen, was die Auswahl des „richtigen“ CPPs für eine Anwendung erschwert. Ziel dieser vorliegenden Arbeit war es, das pro-apoptotische Peptid KLA mittels CPPs spezifisch in Krebszellen zu transportieren. Untersucht wurden: (I) Verschiedene CPPs in unterschiedlichen Zelltypen zur Selektion der „besten“ CPPs; (II) Der Einflusses des CPP C-Terminus auf die Internalisierung und zelluläre Verteilung; (III) Der zelluläre KLA-Transport mittels CPPs via einer nicht-kovalenten Administration. 22 verschiedene CPPs wurden in sieben Zelltypen untersucht, wobei Toxizität, Zellaufnahme und zelluläre Lokalisation mittels Fluorescein markierten CPPs in Fluoreszenzspektroskopie und konfokaler Mikroskopie betrachtet wurden. Abhängig von der Zellaufnahme wurden die CPPs in drei Gruppen klassifiziert. Die Untersuchung carboxylierter und carboxyamidierter CPP C-Termini ergab, dass in den meisten Fällen ein Carboxyamid die zelluläre Aufnahme begünstigte. Drei CPPs (MPG, Penetratin und Integrin) wurden ausgewählt, um das pro-apoptotische KLA Peptid in zwei Krebszelllinien (MCF-7 Brustkrebszellen und leukämische RAW264.7 Makrophagen) im Vergleich zu Fibroblasten (Cos-7) nicht-kovalent zu transportieren. Der erfolgreiche KLA-Transport hing vom CPP, dessen C-Terminus und der Zelllinie ab. Die Analyse der Viabilität nach CPP:KLA Administration ergab, dass MPG-CONH2:KLA (1:2) toxisch für Makrophagen und Brustkrebszellen, aber nicht für Fibroblasten war. Die Toxizität konnte der Apoptose zugeordnet werden. Die vorliegende Arbeit liefert wichtige Informationen über die Auswahl des passenden CPPs für den nicht-kovalenten Transport des pro-apoptotischen KLA-Peptids. / Limitations in a pharmacological therapy are often due to the inability of drugs to overcome the cell membrane and therefore transporting molecules are being used, e.g. cell penetrating peptides (CPPs). Only a few of the over 100 described CPPs have been compared systematically making the choice of “the” CPP for a given application difficult. The goal of the presented work is the CPP mediated delivery of the pro-apoptotic peptide KLA in breast cancer cells as proof of principle for a therapeutical application. Analysed were (I) Different CPPs in various cell types to select the “best” one, (II) The influence of the CPP C-termini on uptake and localisation, (III) The cellular KLA delivery via a non-covalent CPP administration. 22 CPPs were compared in seven cell types thereby looking at toxicity, cellular uptake and subcellular localisation using fluorescein labelled CPPs for fluorescence spectroscopy and confocal microscopy. The resulting uptake information allowed the classification of the CPPs in three main groups. The evaluation of carboxylated and carboxyamidated CPP C-termini revealed that a carboxyamide mostly enhanced the cellular CPP uptake. Three CPPs were selected (MPG, penetratin and integrin) to deliver the pro-apoptotic KLA peptide in two cancer cell lines (breast cancer MCF-7 cells and RAW264.7 macrophages) compared to fibroblasts (Cos-7) via the non-covalent strategy. A successful KLA delivery depended on the applied CPP, its C-terminus and the used cell line. The biological activity of the pro-apoptotic KLA peptide was determined via the cell viability (MTT assay). The co-incubation of MPG-CONH2:KLA (1:2) was able to induce toxicity in breast cancer cells and leukaemic macrophages, but not in fibroblasts. The viability reductions were then assigned to apoptosis. This work provides important information for the choice of an adequate CPP for the pro-apoptotic KLA peptide delivery and presents the advantage of the non-covalent delivery strategy.

Na presente pesquisa estudamos o comportamento do sujeito pronominal no português uruguaio (PU) falado na região de fronteira entre Brasil e Uruguai (Santana do Livramento-Rivera). Associando alguns pressupostos da Teoria de Princípios e Parâmetros (CHOMSKY, 1981), mais especificamente o Parâmetro do Sujeito Nulo (pro-drop), e da Sociolinguística Variacionista (WEINREICH, LABOV e HERZOG, 1968), analisamos uma amostra de língua oral composta de falas de 37 informantes, investigando as seguintes questões: (1) no português uruguaio teríamos uma mudança linguística em curso no que se refere ao Parâmetro do Sujeito Nulo, de modo que ele venha a se tornar um sujeito pronominal pleno?; (2) quais são os fatores linguísticos e sociais condicionantes dessa produção e em que direção vai essa tendência, português brasileiro (PB) ou espanhol (E)? A análise dos fatores sociais e linguísticos selecionados permitiu observar que o PU também passa por um processo de mudança paramétrica, ainda que mais lenta, devido ao fato de estar submetido à pressão simultânea de duas línguas, e se apresenta como uma língua em que, em alguns contextos, a realização do sujeito pronominal é plena e, em outros, ainda conserva as características das línguas pro-drop. Confirma-se, ainda, que essa disputa não está terminada, embora haja indícios nos dados da geração mais nova de que as regras do espanhol vêm ganhando alguma força, talvez pela pressão da escola e dos meios de comunicação. Tudo indica, então, que a base do PU é o PB, porém algumas características do E estão presentes nesse aspecto da gramática. A análise realizada confirma a maior parte das hipóteses iniciais e dá indícios de que o PU e o PB poderiam configurar-se como duas línguas distintas, da mesma forma que o PB e o PE, como aponta Galves (1998), isto é, tudo parece indicar que, mesmo quando os enunciados superficialmente se parecem, a língua interna (Língua-I) seria diferente e os enunciados dos falantes teriam estatutos diferentes, algo que, no entanto, será necessário demonstrar com um outro modelo de coleta de dados, de natureza mais introspectiva. / In this research we study the behavior of the pronominal subject in the Uruguayan Portuguese language (PU) spoken in the border region between Brazil and Uruguay (Santana do Livramento-Rivera). Associating some assumptions from the Theory of Principles and Parameters (CHOMSKY, 1981), specifically the Null Subject Parameter (pro-drop), and from the Variationist Sociolinguistics (WEINREICH, LABOV and HERZOG, 1968), we analyze a sample of spoken language consisting of 37 informants, in order to investigate the following questions: (1) would we have in the Uruguayan Portuguese language a language change in progress with regard to the Null Subject Parameter, so that it will become a full pronominal subject? (2) what are the linguistic factors and social conditions of production and in which direction will this trend, Brazilian Portuguese (BP) or Spanish (E)? The analysis of social factors and selected language programs suggests that the PU is also undergoing a process of parametric change, albeit slower, due to the fact of being subjected to the pressure of two languages simultaneously, and is presented as a language that, in some contexts, the performance of the pronominal subject is full, and in others it still retains the characteristics of pro-drop languages. It is also confirmed that this dispute is not over, although there are indications in the data of the younger generation that the rules of Spanish are gaining some strength, maybe due to the pressure of school and the media. The analysis confirms most of our initial hypothesis and provides evidence that the PU and CP could be framed as two distinct languages, in the same way as the PB and PE, as appointed by GALVES (1998), that is, everything seems to indicate that, even when the statements superficially resemble, the internal language (Language I) would be different and the production of the speakers would have different statutes, something which, however, would just be demonstrated by means of another more introspective model of data collection.

Made available in DSpace on 2016-08-10T10:39:18Z (GMT). No. of bitstreams: 1 PRISCILLA SILVA ROSA DE ALMEIDA.pdf: 3720637 bytes, checksum: b39a9c071d90058d47a3a8c7aec2c7f3 (MD5) Previous issue date: 2008-07-21 / Soft tissue sarcomas (STS) are tumors with mesodermical origin, comprising about 1% of all adult neoplasms. Because of its effect on the p53 protein coding sequence, and its association with an increased risk for some cancer types, TP53 codon 72 polymorphism has been investigated in several studies. TP53 codon 72 codes for either Arginine (p53Arg), or Proline (p53Pro) at the p53 protein primary sequence. It was demonstrated that such amino acid change affects p53 biochemical and biological properties, and several studies have been developed in order to associate TP53 codon 72 polymorphisms as a risk, and as a prognostic factor for different cancer types. Any published study on the TP53 codon 72 polymorphism in adult soft tissue sarcomas was found in the literature. The present study aimed to investigate TP53Arg/Pro polymorphism as a potential prognostic factor in 100 adult subjects with STS. Patients were assisted at the Hospital Araújo Jorge of the Associação de Combate ao Câncer em Goiás in Goiânia, Brazil. DNA from patients was obtained from formaldehyde-fixed and paraffin-embedded tissue samples stored at the Pathology Department of the institution. Control group included 85 healthy donors randomly selected from Goiânia s population and, for this group, DNA extraction was performed from peripheral blood. Polymorphism genotyping was achieved by using polymerase chain reaction (PCR) with specific primer sets for each polymorphic variant. Statistical analysis was performed by using GenePop Ò web version 3.4 software. In this study, TP53 allelic and genotypic frequencies were investigated for subjects and controls, however, any statistical difference between the two groups was found. Our study supports the evidence that p53Arg is the most frequent allele in Latin American population, but worldwide genic and genetic frequency data are conflicting because of ethnical differences among the studied populations. According to the results, no significant association was demonstrated between TP53 codon 72 polymorphism and clinocopathological characteristics such as gender, age, tumor localization, histology, tumor size, stage, grade, node status, and distant metastasis. The five-year overall survival for the study group was 48.1%. Tumors with p53Pro/Pro genotype demonstrated a reduced survival rate (30%) when compared to p53Arg/Arg (45%), and p53Arg/Pro group (54.9%), but this association was not statistically significant (p = 0.444). In the present study, the p53Arg variant was not statistically associated with a more favorable prognosis in adult STS patients. / Os sarcomas de partes moles (SPM) são tumores de origem mesodérmica, representando cerca de 1% do total das neoplasias em adultos. O polimorfismo do códon 72 do gene TP53 é extensivamente estudado por causar impacto na seqüência codificadora do gene, além de estar associado ao maior risco para o desenvolvimento de alguns tipos de câncer. Este polimorfismo resulta na expressão de arginina (p53Arg) e/ou prolina (p53Pro) na posição 72 da proteína p53. As formas polimórficas de TP53, em relação ao polimorfismo do códon 72, apresentam propriedades bioquímicas e biológicas diferentes, e por esta razão, vários estudos foram conduzidos na tentativa de associar tais formas polimórficas como fator de risco e prognóstico para inúmeras neoplasias. Entretanto, a literatura não relata nenhum estudo que associe este polimorfismo aos sarcomas de partes moles do adulto. Neste contexto, o objetivo do presente estudo foi avaliar o polimorfismo p53Arg/Pro como potencial fator de risco e/ou prognóstico em 100 casos de SPMs em adultos atendidos no Hospital Araújo Jorge da Associação de Combate ao Câncer em Goiás. O grupo controle incluiu 85 indivíduos saudáveis selecionados aleatoriamente da população da cidade de Goiânia. As amostras dos casos constituíram de tecidos fixados em formol e incluídos em parafina e, para a extração de DNA, os tecidos foram previamente desparafinizados. A extração de DNA do grupo controle foi realizada a partir de sangue periférico. Para a genotipagem do polimorfismo, a reação em cadeia da polimerase (PCR) foi realizada utilizando conjuntos de primers específicos para cada variante polimórfica. Após a análise dos dados obtidos, verificou-se que as freqüências alélicas e genotípicas não apresentaram diferenças estatisticamente significativas entre os casos e os controles. Nosso estudo corrobora com as evidências de que o alelo p53Arg é o mais comum em populações latinoamericanas. Entretanto, os dados sobre as freqüências gênicas e genotípicas da literatura mundial são conflitantes, fato que pode ser atribuído às diferenças étnicas descritas entre as populações estudadas. Nenhuma relação estatisticamente significativa foi encontrada entre o polimorfismo do códon 72 de TP53 e as características clínico-patológicas estudadas, como sexo, idade agrupada, localização, histologia, tamanho e grau histológico tumoral, estadiamento e presença de mestástases. A sobrevida global em cinco anos para o grupo estudado foi de 48,1%. As análises de sobrevida em relação ao polimorfismo de TP53 revelaram que os pacientes cujos tumores apresentaram o genótipo p53Pro/Pro tiveram sobrevida inferior (30%), quando comparados ao grupo de pacientes com os genótipos p53Arg/Arg (45%) e p53Arg/Pro (54,9%). Entretanto, essa diferença não foi estatisticamente significativa (p = 0,444). Sabese que a isoforma p53Arg apresenta função apoptótica mais marcante. Esta característica pode conferir ao paciente um melhor prognóstico da doença. No presente trabalho, contudo, não pudemos verificar que esta variante esteve associada a um prognóstico mais favorável em pacientes adultos com SPMs.

A l’heure où l’Angola voit dans les ressources en matières premières et dans la démocratie des opportunités pour le développement, il est urgent de faire une halte sur la question des ressources humaines. Un vrai développement passe nécessairement par la promotion de l’homme et de sa culture habité par la dialectique de la vie et de la mort. Notre recherche sur "L’exclusion culturelle et le salut en Jésus-Christ dans le contexte actuel de l’Angola" prend l’homme dans sa condition culturelle et religieuse. Ainsi, face à l’exclusion comme déni de l’humanité de l’autre et du salut promis à tout homme, nous proposons l’inclusion comme puissance salvifique posée par le Messie Jésus et annoncée dans l’Évangile, inclusion à réaliser sur tous les modes : politique, social, religieux et culturel. En proposant l’inclusion comme paradigme du salut, nous soulignons la mission de l’Église qui est essentiellement témoignage actif de Jésus-Christ dont la pro-existence révèle le visage de l’amour inclusif de Dieu compris comme miséricorde et don total de vie. / At a time when Angola looks into the raw material resources and democracy as the opportunities for development, it is urgent to pause on the question of human resources. A true development necessarily means the promotion of the human person and its lived culture through the dialectics of life and death. Our research on "Cultural Exclusion and Salvation in Jesus Christ in the present context of Angola" takes man in his cultural and religious conditions. So, facing exclusion as a denial of the humaneness of the other and the salvation promised to all men, we propose an inclusion as a salvific force laid down by Jesus the Messiah and as announced in the Gospels, the inclusion to realise in all modes : political, social, religious and cultural. In proposing inclusion as a paradigm of salvation, we emphasize on the mission of the Church which is essentially an active witness of Jesus Christ of which the pro-existence reveals the face of inclusive love of God understood as the mercy and a total gift of life.

Na presente pesquisa estudamos o comportamento do sujeito pronominal no português uruguaio (PU) falado na região de fronteira entre Brasil e Uruguai (Santana do Livramento-Rivera). Associando alguns pressupostos da Teoria de Princípios e Parâmetros (CHOMSKY, 1981), mais especificamente o Parâmetro do Sujeito Nulo (pro-drop), e da Sociolinguística Variacionista (WEINREICH, LABOV e HERZOG, 1968), analisamos uma amostra de língua oral composta de falas de 37 informantes, investigando as seguintes questões: (1) no português uruguaio teríamos uma mudança linguística em curso no que se refere ao Parâmetro do Sujeito Nulo, de modo que ele venha a se tornar um sujeito pronominal pleno?; (2) quais são os fatores linguísticos e sociais condicionantes dessa produção e em que direção vai essa tendência, português brasileiro (PB) ou espanhol (E)? A análise dos fatores sociais e linguísticos selecionados permitiu observar que o PU também passa por um processo de mudança paramétrica, ainda que mais lenta, devido ao fato de estar submetido à pressão simultânea de duas línguas, e se apresenta como uma língua em que, em alguns contextos, a realização do sujeito pronominal é plena e, em outros, ainda conserva as características das línguas pro-drop. Confirma-se, ainda, que essa disputa não está terminada, embora haja indícios nos dados da geração mais nova de que as regras do espanhol vêm ganhando alguma força, talvez pela pressão da escola e dos meios de comunicação. Tudo indica, então, que a base do PU é o PB, porém algumas características do E estão presentes nesse aspecto da gramática. A análise realizada confirma a maior parte das hipóteses iniciais e dá indícios de que o PU e o PB poderiam configurar-se como duas línguas distintas, da mesma forma que o PB e o PE, como aponta Galves (1998), isto é, tudo parece indicar que, mesmo quando os enunciados superficialmente se parecem, a língua interna (Língua-I) seria diferente e os enunciados dos falantes teriam estatutos diferentes, algo que, no entanto, será necessário demonstrar com um outro modelo de coleta de dados, de natureza mais introspectiva. / In this research we study the behavior of the pronominal subject in the Uruguayan Portuguese language (PU) spoken in the border region between Brazil and Uruguay (Santana do Livramento-Rivera). Associating some assumptions from the Theory of Principles and Parameters (CHOMSKY, 1981), specifically the Null Subject Parameter (pro-drop), and from the Variationist Sociolinguistics (WEINREICH, LABOV and HERZOG, 1968), we analyze a sample of spoken language consisting of 37 informants, in order to investigate the following questions: (1) would we have in the Uruguayan Portuguese language a language change in progress with regard to the Null Subject Parameter, so that it will become a full pronominal subject? (2) what are the linguistic factors and social conditions of production and in which direction will this trend, Brazilian Portuguese (BP) or Spanish (E)? The analysis of social factors and selected language programs suggests that the PU is also undergoing a process of parametric change, albeit slower, due to the fact of being subjected to the pressure of two languages simultaneously, and is presented as a language that, in some contexts, the performance of the pronominal subject is full, and in others it still retains the characteristics of pro-drop languages. It is also confirmed that this dispute is not over, although there are indications in the data of the younger generation that the rules of Spanish are gaining some strength, maybe due to the pressure of school and the media. The analysis confirms most of our initial hypothesis and provides evidence that the PU and CP could be framed as two distinct languages, in the same way as the PB and PE, as appointed by GALVES (1998), that is, everything seems to indicate that, even when the statements superficially resemble, the internal language (Language I) would be different and the production of the speakers would have different statutes, something which, however, would just be demonstrated by means of another more introspective model of data collection.

The aim of this paper is to study whether there is a correlation between how subject pronouns are introduced in Spanish teaching and the pupils' difficulties regarding both their use and the understanding of verb conjugation. What is being investigated is: a) to what extent and in what contexts subject pronouns appear in Spanish textbooks, and b) how secondary education teachers of Spanish work with subject pronouns in the classroom. My conclusion is that the subject pronouns are under-represented in Spanish textbooks, probably due to the interpretation of the null subject parameter and Spanish as a pro-drop language. As far as the work with subject pronouns in the classroom is concerned, a more in-depth analysis needs to be carried out in order to be able to demonstrate a reliable correlation between the teaching method and the students' results. / Syftet med detta arbete är att studera om det finns ett samband mellan hur subjektspronomina introduceras i spanskaundervisningen och elevernas svårigheter kring både användandet av dessa och förståelsen för verbböjning. Det som undersöks är: a) i vilken utsträckning och i vilka sammanhang förekommer subjektspronomina i läromedel för undervisning i spanska, och b) hur högstadielärare i spanska arbetar med subjektspronomina i klassrummet. Min slutsats är att subjektspronomina finns underrepresenterade i läroböcker i spanska, förmodligen på grund av den tolkning som görs av nollsubjektsparametern och spanska som pro-drop språk. Vad gäller arbetet med subjektspronomina i klassrummet behöver en mer fördjupad analys genomföras för att det ska kunna påvisas ett tillförlitligt samband mellan undervisningsmetod och elevernas resultat.

This work offers a complete critical overview of the samizdat magazine Sado-Maso1 published in Prague in the years 1983-1986. I tried to make a complete overview and characteristics of the magazine - the main themes of each issue, historical, ideological and literary context of publishing of the Sado-Maso magazine. I also mention publications which were connected with the S-M magazine - independent movies, other samizdat magazines and books. In the Attachment there is article bibliography (containing indexes), covers of the Sado-Maso individual issues, interviews with authors and conterporary witnesses, copies of ,StB' documentary where Sado-Maso magazine was mentioned connected with Klín and Satan Actions. Egon Bondy's essay Zaplaťpánbů za "underground" published exclusively in Sado- Maso Nr. 7 ½ is presented here in commented edition. The goal of this work is to provide information about a not well-known part of Czechoslovak underground culture - a part which still has not been paid much attention on the side of literary history. 1 The title ,Sado-Maso' and the abreviation ,S-M' are used in the same meaning. Keywords SAMIZDAT, MAGAZINES, SADO-MASO, JEN PRO BLÁZNY, OPIUM PRO LID, (ZEN)BUDDHISM, ECOLOGY, ANARCHISM, PSYCHOLOGY, PUNK, MOVIES OF THE SO-CALLED INDEPENDENT PRODUCTION, VISUAL ARTS,...

Dentre as variações morfológicas possivelmente associadas ao potencial fisiológico das sementes está o tamanho do embrião. Uma das abordagens atuais da pesquisa sobre análise de imagens de sementes radiografadas é verificar até que ponto a relação entre o grau de desenvolvimento embrionário e o espaço disponível na cavidade interna da semente está associada ao desempenho da semente. Os objetivos deste trabalho foram verificar a eficiência de diferentes testes na avaliação do vigor de sementes de algodão ao longo do armazenamento, verificar a possível relação entre o espaço livre na cavidade interna e o potencial fisiológico das sementes durante o armazenamento e verificar se há relação entre o espaço livre e o desempenho de sementes de algodão. Diferentes lotes de sementes de algodão, cultivares BRS 293 e FMT 709, foram armazenados em câmara fria e seca (10 °C e 30% de UR do ar) e em ambiente natural, durante oito meses. As avaliações da germinação e do vigor (primeira contagem de germinação, germinação a baixa temperatura, envelhecimento acelerado tradicional e com solução saturada de NaCl, emergência de plântulas em campo e análise computadorizada de imagens de plântulas - SVIS®) foram realizadas a cada dois meses de armazenamento. Para avaliação do espaço livre, as sementes de cada lote foram radiografadas e analisadas por meio dos softwares Tomato Analyzer e Image-Pro® Plus. Os resultados indicaram que os testes de envelhecimento acelerado (tradicional e com solução saturada de NaCl), emergência de plântulas em campo e SVIS® (comprimento de plântulas) são adequados para avaliação do vigor de sementes de algodão, sendo o envelhecimento acelerado tradicional e as análises SVIS® (comprimento de plântulas) sensíveis para detectar redução no potencial fisiológico das sementes durante o armazenamento. No entanto, a queda do vigor das sementes de algodão não está associada ao espaço livre na cavidade interna das sementes. Contudo, há indícios de uma relação inversamente proporcional entre o espaço livre e a germinação de sementes de algodão. / One of the morphological variations usually referred as associated to seed physiological potential is the embryo size. A primary interest of image analysis of Xrayed seeds is to identify whether the ratio between the degree of embryo development and the available space in the internal seed cavity is related to seed performance. The objectives of this research were to verify the accuracy of different tests to assess the vigor of cotton seeds during storage, to check the possible relationship between the free space and cotton seed physiological potential during storage and to verify if the relationship between the free space and cotton seed performance exist. Cotton seed lots of BRS 293 and FMT 709 cultivars were stored during eight months under two environments: cold and dry chamber (10 °C and 30% relative air humidity) and laboratory non-controlled condition. Evaluations of seed germination and vigor (germination first count, cool germination, traditional and saturated salt accelerated aging, field seedling emergence and seedling imaging automated analysis - SVIS®) were performed at two months intervals during storage. Free space was evaluated in radiographed seeds from each lot using Tomato Analyzer and Image-Pro® Plus softwares. Results showed that the accelerated aging tests (traditional and with saturated NaCl), field seedling emergence and SVIS® (seedling length) are suitable for evaluate the cotton seed vigor and the traditional accelerated aging and SVIS® (seedling length) are sensitive to detect reductions in seed physiological potential during storage. However, the reduction of cotton seed vigor is not associated with the free space in the internal cavity of the seed. Nevertheless, there is evidence of an inverse relationship between the free space and germination of cotton seed.

- PROCEDURAL STATUS OF INTERNATIONAL NON-GOVERNMENTAL ORGANISATIONS BEFORE INTERNATIONAL HUMAN RIGHTS COURTS The position of international NGOs within the system of international law or specifically their position before individual international bodies is a topic that the international law theory does not consider very much. Hence, this master's thesis aims to present what role do the international NGOs play in the international arena and particularly places the focus on the procedural roles they may take upon in the european and the inter-american human rights systems. The thesis is divided into 3 main parts and several chapters. The first part deals with the history of international NGOs and delimiting the term. Regarding the latter several definitions are presented and assessed and the differences between the international NGOs and transnational corporations are drawn. The thesis then explores the position of international NGOs as subjects of international law and their role in human rights protection which reflects the topis of the thesis. The second part is devoted to particular roles of international NGOs before the selected courts. The first chapter deconstructs the institute of amicus curiae including the way it is embeded into the systems of both courts and the way it is made use of. This...

La maladie de Crohn (MC) est une maladie inflammatoire chronique du tube digestif caractérisée par un état d’hyperactivation du système immunitaire intestinal. L’étiologie de cette maladie est encore mal connue mais les données cliniques et expérimentales montrent que l’étiologie de la MC fait intervenir des facteurs environnementaux, génétiques et infectieux entraînant une réponse immunitaire aberrante. A ce jour, aucun traitement spécifique n’est disponible et la MC représente un problème majeur de santé publique de par ses conséquences invalidantes, sa nature chronique et récidivante, et sa prévalence croissante. Plusieurs groupes dont le nôtre ont rapporté une colonisation anormale de la muqueuse intestinale des patients atteints de MC par des souches de E. coli adhérentes et invasives dénommées AIEC. La caractérisation des souches AIEC a montré qu’elles sont capables d’adhérer et d’envahir des cellules épithéliales intestinales, de survivre et se multiplier dans des macrophages et d’induire une forte réponse pro-inflammatoire. Des études récentes ont montré que l’autophagie est induite dans les cellules hôtes en réponse à l’infection par les AIEC afin de contrôler la réplication intracellulaire des AIEC.Les travaux menés au cours de cette thèse visent à étudier les mechanismes moleculaires impliqués dans l’interaction entre l’hôte et les AIEC avec 2 objectifs principaux : (i) étudier l’implication des exosomes dans les interactions hôte/AIEC, (ii) étudier l’implication de la voie de signalisation GCN2/eIF2α/ATF4 dans la réponse autophagique à l’infection par les AIEC. Concernant le premier axe de recherche, les résultats ont permis de mettre en lumière une fonction jusque-là inconnue des exosomes dans la communication cellule à cellule au cours des réponses de l’hôte à l’infection par les AIEC. Ainsi, nous avons mis en évidence que les exosomes sont impliqués à la fois dans la réplication intracellulaire des AIEC et dans les réponses immunitaires innées des cellules hôtes à l’infection. Les résultats du deuxième axe de recherche ont permis de révéler un rôle essentiel de la voie de signalisation GCN2/eIF2α/ATF4 dans la réponse autophagique de l’hôte à l’infection par les AIEC associés à la MC. Nous avons démontré que l’infection par les souches AIEC conduit à l’activation de cette voie de signalisation, entraînant la mise en place d’une machinerie autophagique efficace pour contrôler la réplication intracellulaire des AIEC et inhiber l’inflammation. / Crohn's disease (CD) is a chronic inflammatory disease of the digestive tract characterized by a state of hyperactivation of the intestinal immune system. The etiology of this disease is still poorly understood, but clinical and experimental data show that the etiology of CD involves environmental, genetic and infectious factors leading to an aberrant immune response. To date, no specific treatment is available and CD represents a major public health problem due to its disabling consequences, its chronic and recurrent nature, and its increasing prevalence. Several groups, including ours, have reported abnormal colonization of the intestinal mucosa of CD patients with adherent and invasive E. coli strains called AIEC. The characterization of AIEC strains has shown that they are able to adhere and invade intestinal epithelial cells, survive and multiply in macrophages and induce a strong pro-inflammatory response. Recent studies have shown that autophagy is induced in host cells in response to AIEC infection in order to control the intracellular replication of AIEC. The work conducted during this thesis aims to investigate the molecular mechanisms involved in interaction between host and AIEC with 2 main objectives: (i) to study the involvement of exosomes in host / AIEC interactions, (ii) to study the involvement of the GCN2 / eIF2α / ATF4 signaling pathway in the response autophagic to infection with AIEC. With regard to the first line of research, the results shed light on a hitherto unknown function of exosomes in cell-to-cell communication during host responses to AIEC infection. Thus, we have shown that exosomes are involved in both intracellular replication of AIECs and innate immune responses of host cells to infection. The results of the second line of research revealed a critical role for the GCN2 / eIF2α / ATF4 signaling pathway in the host autophagic response to AIEC infection associated with CD. We have shown that infection with AIEC strains leads to activation of this signaling pathway, leading to the establishment of autophagic machinery that is effective in controlling intracellular replication of AIEC and inhibiting inflammation.