Les objectifs de cette thèse sont d'analyser l'influence des processus humides sur les dépressions des moyennes latitudes en s'intéressant plus particulièrement aux déplacements des dépressions et à la formation des vents forts près de la surface. Ces recherches ont été effectuées à l'aide de simulations idéalisées du modèle de méso-échelle Méso-NH. La première étude s'est focalisée sur l'effet des processus humides sur le déplacement des dépressions dans la direction perpendiculaire à l'axe du courant-jet, c'est à dire principalement le déplacement vers les pôles. Le mécanisme expliquant ce déplacement est dû à l'advection non-linéaire de la dépression de surface par les anomalies cyclonique et anticyclonique d'altitude se trouvant de part et d'autre de la dépression de surface. En présence d'humidité, l'anomalie anticyclonique d'altitude se renforce du fait du dégagement de chaleur latente dans les zones d'ascendances, et ainsi le déplacement perpendiculairement à l'axe du jet et vers les pôles s'accélère. On montre aussi que le déplacement vers l'est des dépressions s'accélère en incluant les processus humides. La seconde étude porte sur la formation des vents forts près de la surface et notamment sur ce qu'on appelle les "sting jets". Les "sting jets" sont des jets dont les masses d'air associées descendent rapidement du milieu de la troposphère au niveau de la tête du nuage jusqu'au sommet de la couche limite et peuvent déclencher des vents dévastateurs en surface. Ceux-ci se forment lorsque le front chaud se déplace à l'arrière de la dépression et qu'une fracture frontale apparaît. Les simulations montrent que, lorsqu'une dépression traverse l'axe du courant-jet de son côté chaud à son côté froid, le retour en arrière du front chaud se produit tandis qu'une dépression naissant côté froid du courant-jet possède un front chaud beaucoup moins actif. Ainsi, la formation d'un " sting jet " n'apparaît que dans le premier cas , moyennant une résolution verticale élevée. C'est la présence d'un forçage géostrophique dans un environnement neutre vis-à-vis de l'instabilité symétrique qui facilite la descente des masses d'air et la formation du " sting jet ". / The objective of this thesis is to analyze the influence of moist processes on mid-latitude cyclones, and specifically the tracks of cyclones and the formation of strong winds near the surface. These researches have been made using idealized simulations of the mesoscale model Méso-NH. The first study focused on the effects of moist processes on the motion perpendicularly to the zonal jet axis, poleward. The main mechanism explaining this motion involves the nonlinear advection of the surface cyclone by the upper-level cyclonic and anticyclonic perturbations located on both sides of the surface cyclone. In the presence of humidity, the upper-level anticyclonic anomaly strengthens due to the latent heat release in the ascent zones, and the poleward motion is reinforced. The eastward motion of the cyclone is also reinforced with the inclusion of moist processes. The second study focuses on the formation of near-surface strong winds which are triggered by the so-called " sting-jets ". These jets correspond to descending air masses from the mid-troposphere near the cloud head down to the top of the boundary layer, and can trigger devastating surface winds. In the case where synoptic perturbations are initialized south of the baroclinic zonal jet, the bent-back warm front phase occurs, whereas when they are initialized on the zonal jet axis, the surface cyclone has a less active warm front. Thus, the formation of a sting-jet only occurs in the first case, and with a high vertical resolution. It is the geostrophic forcing in a globally neutral environment relative to conditional symmetric instability which facilitates the airstreams descent and the formation of the sting-jet.

Les inhibiteurs de poly(ADP-ribose) polymérase (PARPi) ciblent sélectivement les cellules porteuses de défauts des voies de réparation de l’ADN tels que les mutations de BRCA1/2 et les défauts d’ERCC1. Sur le plan clinique, plusieurs PARPi ont été approuvés pour le traitement des cancers BRCA-mutés ou platine-sensibles du sein et de l’ovaire, et des essais cliniques sont en cours pour évaluer l’efficacité des PARPi dans le cancer bronchique non-à-petites cellules (CBNPC) platine-sensible. Alors que les PARPi ont un fort potentiel thérapeutique dans les cancers comportant des défauts de réparation de l’ADN, de plus en plus d’essais cliniques évaluent également l’efficacité de ces médicaments en combinaison avec les « inhibiteurs d’immune checkpoints » (ICI) dans diverses populations de patients. Dans ce contexte, il est essentiel de mieux comprendre comment les PARPi modulent la réponse immunitaire anti-tumorale, et d’étudier le potentiel immunologique inhérent de ces médicaments.Dans cette étude, nous avons établi que les cellules de CBNPC déficientes en ERCC1 expriment fortement la signature interféron (IFN) de type I, et que les tumeurs de CBNPC ayant une faible expression d’ERCC1 ont un infiltrat lymphocytaire renforcé. En utilisant des lignées cellulaires isogéniques et des xénogreffes dérivées de patients, nous avons montré que plusieurs PARPi, notamment l’olaparib et le rucaparib, ont des propriétés immunomodulatrices dans les modèles de CBNPC ERCC1-déficients et de cancers du sein triple-négatifs (CSTN) BRCA1-mutés. D’un point de vue mécanistique, les PARPi génèrent des fragments d’ADN cytoplasmiques ayant les caractéristiques de micronoyaux ; ceux-ci activent la voie cGAS/STING et déclenchent une réponse IFN de type I, associée à la sécrétion de la cytokine CCL5. De manière importante, ces effets sont largement diminués dans les cellules de CSTN BRCA1-révertantes et les cellules de CBNPC ré-exprimant ERCC1, ce qui suggère que les défauts de réparation de l’ADN amplifient les phénotypes immunitaires associés au traitement par PARPi. En outre, ces effets sont totalement abrogés dans les cellules de CSTN PARP1-neutralisées, ce qui confirme que les phénotypes observés dépendent d’un effet spécifique des PARPi sur leur cible.Au-delà de leur potentiel d’activation d’une immunité spécifique des cellules cancéreuses via cGAS/STING et la signalisation IFN de type I, nous avons également constaté que les PARPi potentialisent les effets inducteurs de l‘IFN de type II sur l’expression de PD-L1 dans des lignées cellulaires et cellules tumorales fraîches de patients CBNPC, surtout en présence de défauts d’ERCC1. De plus, nous avons montré que certains PARPi, utilisés à des concentrations létales, activent de manière indépendante les éléments moléculaires clés de la mort cellulaire immunogénique, dont l’exposition de la calréticuline à la surface des cellules cancéreuses, la sécrétion d’ATP et le relargage d’HMGB1 en grandes quantités dans le milieu extracellulaire.Dans l’ensemble, ces données précliniques suggèrent que les PARPi ont des propriétés immunomodulatrices intrinsèques qui participent à l’activation de réponses immunitaires anti-tumorales ; ce potentiel pourrait être exploité cliniquement en combinaison avec les ICI dans des populations adéquatement sélectionnées au plan moléculaire. / Poly(ADP-ribose) polymerase inhibitors (PARPi) selectively target cancer cells with DNA repair deficiencies such as BRCA1/2 mutations or ERCC1 defects. Clinically, several PARPi are currently approved for the treatment of BRCA-mutant or platinum-sensitive advanced ovarian and breast cancers, and ongoing clinical trials are investigating the efficacy of PARPi in platinum-sensitive Non-Small Cell Lung Cancer (NSCLC). While PARPi constitute potent targeted therapies for the treatment of DNA repair-deficient malignancies, an increasing number of clinical trials are also evaluating their efficacy in combination with immune checkpoint inhibitor (ICI) in various populations. In this context, it is of critical importance to better understand how PARPi might modulate immune responses against cancer, and to investigate the inherent immunological potential of these agents.In this study, we show that ERCC1-defective NSCLC cells exhibit an enhanced type I interferon (IFN) transcriptomic signature and that low ERCC1 expression correlates with increased lymphocytic infiltration in human NSCLC tumours. Using isogenic cell lines and patient-derived xenografts, we further demonstrate that several clinical PARPi, including olaparib and rucaparib, display cell-autonomous immunomodulatory properties in ERCC1-defective NSCLC and BRCA1-mutant triple-negative breast cancer (TNBC) models. Mechanistically, PARPi generate cytoplasmic chromatin fragments with micronuclei characteristics; this activates the cGAS/STING pathway and elicits downstream type I IFN signalling and CCL5 secretion. Importantly, these effects are suppressed in BRCA1-reverted TNBC cells and ERCC1-rescued NSCLC cells, suggesting that DNA repair defects exacerbate the innate immunity-related phenotypes triggered by PARPi. Similarly, these effects are totally abrogated in PARP1-null TNBC cells, supporting the on-target effect of PARPi in mediating such phenotypes. Besides this potential to activate tumour cell-autonomous immunity through cGAS/STING and type I IFN signalling, we also observed that PARPi synergize with type II IFN to induce PD-L1 expression in NSCLC cell lines and fresh patient tumour cells, especially in the ERCC1-deficient setting. Moreover, we show that lethal concentrations of some PARPi independently activate the key damage-associated molecular patterns dictating the immunogenicity of cancer cell death, including calreticulin exposure at the tumour cell surface, ATP secretion and HMGB1 release in the extracellular compartment.Together, these preclinical data suggest that PARPi have intrinsic immunomodulatory properties that activate anti-cancer immune responses; this could be exploited clinically in combination with ICI in appropriately molecularly-selected populations.

HINTERGRUND: Eine inadäquate Aktivierung von Typ-1-IFN kann in der Entstehung von Autoimmunität und Autoinflammation resultieren. Die einer solchen dysregulierten Typ-1-IFN-Achse zu Grunde liegenden Störungen werden primär über das angeborene Immunsystem vermittelt. Krankheitsbilder, die durch eine chronische Typ-1-IFN-Aktivierung bedingt sind, werden daher unter dem Begriff Typ-1-Interferonopathien zusammengefasst. Diese Gruppe seltener, genetisch bedingter Erkrankungen zeichnet sich durch eine große symptomatische Bandbreite aus, wobei vor allem neurologische und kutane Manifestationen im Vordergrund stehen. Bisher aufgeklärte Pathomechanismen dieser systemisch-entzündlichen Erkrankungen haben einen Einblick in neue zellintrinsische Mechanismen gegeben, die zu einer Typ-1-IFN-Aktivierung führen und auf Störungen im Metabolismus und der immunologischen Erkennung intrazellulärer Nukleinsäuren beruhen. Daraus ergeben sich auch Erkenntnisse hinsichtlich des Einsatzes einer immunmodulatorischen Therapie zur kausalen Behandlung von Typ-1-Interferonopathien. FRAGESTELLUNG: Typ-1-Interferonopathien gehören zu den genetisch bedingten, seltenen Erkrankungen. Eine Diagnosestellung ist jedoch aufgrund mangelnder Kenntnisse über die Krankheitsursache häufig nicht möglich, sodass kausale Therapieansätze für viele Patienten nicht existieren. Die Aufklärung der genetischen Ursache solcher Erkrankungen ist demnach von essentieller Bedeutung. Auch die hier untersuchten Familien leiden an bisher nicht molekulargenetisch diagnostizierten Krankheiten, deren Phänotyp jedoch eine Typ-1-Interferonopathie vermuten lässt. Vor diesem Hintergrund war es das Ziel dieser Arbeit, bisher unbekannte Gene zu identifizieren und zu charakterisieren, welche Krankheitsbilder mit chronischer Typ-1-IFN-Aktivierung verursachen. MATERIAL UND METHODEN: Anhand klinischer Daten wurden zunächst die Krankheitsbilder der zwei betroffenen Familien charakterisiert und hinsichtlich der Gemeinsamkeiten mit bekannten Typ-1-Interferonopathien untersucht. Zudem wurde das Vorliegen einer chronischen Typ-1-IFN-Aktivierung bei den erkrankten Familienmitgliedern untersucht. Mit Hilfe von Exomanalysen und anschließenden bioinformatischen Analysen wurde nach dem ursächlichen Krankheitsgen gesucht. Die in diesem Rahmen in der Familie 1 nachgewiesene STING-Mutation wurde unter Verwendung von molekularbiologischen sowie zellbiologischen Analysen eingehend untersucht und zudem ihre Konsequenzen auf die IFN-Achse und die Stabilität des STING-Dimers mittels Strukturmodellierungen charakterisiert. Hinsichtlich der Aufklärung der genetischen Ursache der Erkrankung der Familie 2 wurde ebenfalls eine Exomanalyse durchgeführt. Da dabei jedoch keine Mutation identifiziert werden konnte, erfolgte eine Analyse differentiell exprimierter Transkripte mit Hilfe von Transkriptomdaten von Fibroblasten gesunder Kontrollen im Vergleich zu erkrankten Familienmitgliedern der Familie 2. ERGEBNISSE: Im Verlauf dieser Arbeit konnte in der Familie 1 mit einem familiären Chilblain Lupus eine kausale heterozygote Mutation im STING-Gen identifiziert werden. Diese bisher nicht beschriebene Mutation bedingt den Aminosäureaustausch Gly166Glu (G166E) im hochkonservierten Dimerinterface des STING-Proteins. Strukturmodelle ergaben Hinweise auf strukturverändernde Eigenschaften der Mutation G166E, die das STING-Dimer konstitutiv aktivieren. Dies konnte experimentell bestätigt werden, da in den peripheren Blutzellen der Patienten eine erhöhte IFN-Signatur nachgewiesen werden konnte und Patientenfibroblasten eine erhöhte Produktion von IFN-β sowie eine vermehrte Phosphorylierung von IRF3 zeigten. Um den Einfluss einer therapeutischen Immunmodulation über eine JAK-Inhibition zu untersuchen, wurden zwei Erkrankte der Familie 1 mit dem JAK-Inhibitor Tofacitinib über einen Zeitraum von 17 Tagen behandelt. Es konnte dabei in vivo eine signifikant reduzierte IFN-Signatur nachgewiesen werden. Mit Hilfe der Exomanalyse konnte in der konsanguinen Familie 2 mit einem AGS-ähnlichen Phänotyp keine kausale Mutation identifiziert werden. Es zeigte sich zudem, dass in den Patientenzellen keine erhöhte IFN-Signatur sowie keine erhöhte IFN-β Produktion nachweisbar waren. Die vergleichende Transkriptomanalyse ergab keine auffällige differentielle Expression von Genen des Aminosäure-Metabolismus, der Seneszenz, des Zellzyklus, der DNA-Schadensantwort und DNA-Reparatur sowie von Genen immunologischer Prozesse. Allerdings wurde die vollständig fehlende Expression der Gene COL6A1 und COL6A2 in den Patientenzellen nachgewiesen. SCHLUSSFOLGERUNG: Die identifizierte STING-Mutation der Familie 1 führt zu einer chronischen Aktivierung der Typ-1-IFN-Achse und ist folglich als Gain-of-function Mutation anzusehen. Die Behandlung mit dem immunmodulierenden JAK-Inhibitor Tofacitinib führt zu einer Verminderung der IFN-Signatur und bietet somit einen vielversprechenden Ansatz für eine kausale Therapie. Weiterführende Untersuchungen, vor allem bezüglich der Langzeiteffekte, sind hier jedoch noch erforderlich. Die Erkrankung der Familie 2 ist dagegen vermutlich nicht den Typ-1-Interferonopathien zuzuordnen. Vielmehr scheint es zum jetzigen Zeitpunkt wahrscheinlich, dass eine Mutation innerhalb eines regulatorischen Elements, das die Expression der Gene COL6A1 und COL6A2 reguliert, für den Phänotyp der Erkrankten verantwortlich ist. Weiterführende Untersuchungen sind hier zukünftig von großem Interesse.

Nesta dissertação pretendo reler o debate entre Ronald Dworkin e o positivismo jurídico. Farei isto sob o prisma da filosofia analítica, especificamente, contextualizando o debate no âmbito de uma discussão travada entre três teorias semânticas específicas: a descricional, o externalismo semântico e o bi-dimensionalismo ambicioso. Há algum tempo Dworkin lançou uma crítica ao positivismo, qual seja, o positivismo jurídico pretende reduzir a forma direito de como as coisas são à conformação puramente descritiva de como o mundo é. Disse, ainda, que somente quando concebido como uma teoria semântica é que o positivismo jurídico tornar-se-ia inteligível. Os posivistas, a seu turno, argumentam que a Jurisprudência analítica é um projeto teórico pelo direito e não pelo significado do termo direito e que, portanto, deveríamos manter separados dois tipos de questionamentos: O que é direito? e O que é direito?. Se tudo correr bem, ao reler o debate entre os positivistas e Ronald Dworkin a partir do instrumental obtido no âmbito da teoria semântica, poderemos perceber que pode ser verdade que o positivismo jurídico, enquanto projeto teórico, é sobre o direito, o referente, e não sobre o direito, o termo; entretanto, a forma pela qual o positivismo concebe o questionamento O que é direito?, ele mesmo, parece acabar por qualificá-lo, num sentido não trivial, como semântico. / I intend to reread the debate between Ronald Dworkin and legal positivism. I will do that through the prism of analytic philosophy, specifically in the context of the debate between three specific semantic theories: descriptional, externalism and the ambitious bidimensionalism. Dworkin criticized legal positivism: the legal positivism aims to reduce the law-way of things to the purely descriptive form of the world. He also said that only when conceived as a semantic theory is that legal positivism would become intelligible. The posivists argue that analytical Jurisprudence is a theoretical project about law and not about the meaning of \"law\", therefore we should keep separated two types of questions: \"What is law?\" and \"What is \'law\'?\". If all goes well, when rereading the debate through the prism of the discussion in the context of semantic theories, we will realize that it may be true that legal positivism is about law, the referent, and not about \"law\". However, the way in which positivism conceives the question \"What is law?\" seems to qualify it as semantic in a nontrivial sense.

Made available in DSpace on 2014-06-11T19:30:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-04-20Bitstream added on 2014-06-13T18:41:14Z : No. of bitstreams: 1 junqueira_mep_dr_botfm.pdf: 683998 bytes, checksum: e97592029abff7e2b2e4961aed2d337c (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Universidade Estadual Paulista (UNESP) / Gap / Nossos estudos objetivaram caracterizar as respostas imune inata e específica induzidas pelas peçonhas do muco e do ferrão do bagre Cathorops agassizii . A coleta dos espécimes foi realizada no complexo Baía-Estuário de Santos e São Vicente, localizado no litoral sul do Estado de São Paulo. As peçonhas (do Muco e do Ferrão) apresentaram perfil eletroforético similar entre si. Induzida a inflamação aguda em um modelo experimental murino, as peçonhas apresentaram igualmente a capacidade de induzir aumento da permeabilidade vascular e também edema de pata. A detecção de Leucotrieno B4 e Prostaglandina E2 no lavado da cavidade peritoneal dos camundongos injetados, com ambas as peçonhas, corroboram esta hipótese. Nossos resultados através da microscopia intravital mostraram que as peçonhas induzem um grande número de leucócitos rolantes nas vênulas pós-capilares com focos de extravasamento leucocitário, principalmente de neutrófilos seguido pelo influxo de macrófagos. Além disso, a peçonha do Ferrão induziu uma resolução mais rápida do influxo leucocitário ao contrário da peçonha do Muco que manteve o infiltrado macrofágico por até 7 dias. De maneira interessante, somente a citocina IL-6 foi detectada no lavado peritoneal induzida principalmente pela peçonha do Muco e as quimiocinas KC e MCP-1, por ambas as peçonhas, expressando naquele momento, a participação destes mediadores no recrutamento de neutrófilos e macrófagos para o sítio da lesão. As peçonhas foram eficazes ao induzir uma produção primária e secundária de anticorpos das classes IgM e IgG anti-venenos. Observamos ainda, uma especificidade dos anticorpos produzidos para os componentes das próprias peçonhas e também uma reatividade antigênica cruzada entre elas. / Our studies aimed to characterize the innate and specific immune responses induced by poisons of the mucus and sting of the catfish Cathorops agassizii. The collection of specimens was accomplished in the complex Bay-Estuary System of Santos and São Vicente located in the south coast of the São Paulo State. Poisons of the Mucus and Sting presented similarities in their electrophorectical profiles. In basis of an Induced process of inflammation throughout an experimental murine model, both poisons equally showed the capacity to increase the vascular permeability and also paw edema. The Leucotrien B4'S and Prostaglandine E2 detection in washings of mice peritoneal cavity injected with both poisons corroborates this hypothesis. Our results through an intravital microscopy procedure also showed that the poisons induced a great number of rolling leukocytes in the post-capillary venules with focus on the leukocyte overflow mainly neutrophiles followed by macrophage influx. Besides, the sting poison Sting induced a faster resolution of the leukocyte influx, as well as, the Mucus poison unlike maintained a macrophage infiltrated for up to 7 days. In an interesting way, only the cytokine IL-6 was detected in peritoneal washings induced mainly by the Mucus poison although Quimiokinins KC and MCP-1 were for both poisons, expressing on that moment the participation of these mediators in the neutrophile and macrophage recruitment for the lesion site. The poisons were also effective in the induction of a primary and secondary production of IgM and IgG classes of antibodies. In this research we still observed a specificity of the antibodies produced for the components of the own poisons and also a crossed antigenic reactivity among them.

Les interférons de type I (IFN I) sont des cytokines antivirales aux propriétés puissantes. L’induction, la transmission et la résolution de la réponse immunitaire engendrée par les IFN I est minutieusement régulée. Le concept d’interféronopathie de type I, récemment individualisé par notre équipe, repose sur l’hypothèse que certaines pathologies seraient secondaires au déséquilibre de ces voies de signalisation complexes et à la sécrétion excessive et inappropriée d’IFN I. L’inhibition de celle-ci par des thérapeutiques ciblées permettrait de valider cette hypothèse, si les symptômes allégués s’amélioraient, voire disparaissaient. Ce travail de thèse s’est initialement concentré sur la caractérisation clinique et biologique des interféronopathies monogéniques et polygéniques, et secondairement sur l’identification moléculaire de nouvelles mutations du gène TMEM173 à l’origine de l’interféronopathie liée à STING, également appelée SAVI (STING associated vasculopathy with onset in infancy), syndrome auto-inflammatoire associant une atteinte sévère cutanée et pulmonaire. De nouvelles techniques ont permis la sélection de patients présentant une augmentation de l’IFN I en comparaison à des contrôles sains : la signature IFN I, qPCR de 6 gènes stimulés par l’IFN (IFN stimulated genes – ISGs) et le dosage d’IFN alpha sérique ou plasmatique par méthode du SIMOA (single molecule array) permettant la détection de molécules d’IFN de l’ordre du femtogramme (10-18g). Ces méthodes nous ont ainsi permis d’élargir le spectre clinique phénotypique des interféronopathies de type I, initialement considéré comme essentiellement neurologique. Les patients atteints du syndrome d’Aicardi-Goutières, première interféronopathie monogénique décrite, présentaient les signes suivants : dystonie, spasticité, décalage des acquisitions, calcifications intra-cérébrales et anomalies de la substance blanche. Cependant, l’utilisation systématique de nos méthodes de criblage associée à l’avènement des technologies de séquençage à haut débit (next generation sequencing – NGS) a permis de révéler un phénotype plus large, caractéristique des interféronopathies de type I : sur le plan cutané (engelures, vascularite nécrosante des extrémités, sclérodermie), pulmonaire (pneumopathie interstitielle isolée ou non), musculo-squelettique (arthralgies, arthrites, arthropathie de Jaccoud, myalgies et myosites), ophtalmologique (glaucome), néphrologique (néphropathies lupiques), gastro-entérologique (maladies inflammatoires chroniques intestinales précoces), associées à de l’auto-immunité ou un déficit immunitaire inconstants. Notre méthode de sélection nous a notamment permis d’identifier des patients présentant de manière variable des signes cardinaux de SAVI et une de trois nouvelles mutations activatrices dans une région spécifique du gène TMEM173 (codant pour STING). Ces mutations circonscrivent une région de la protéine à ce jour encore jamais impliquée dans le contrôle de la voie de l’IFN I. STING est une protéine du réticulum endoplasmique qui agit comme adaptateur cytosolique de senseurs intracellulaires d’ADN viral dans une voie de signalisation de l’IFN I. STING active TBK1 (TANK-binding kinase) et permet la transcription des IFN I par la phosphorylation d’IRF3. La Janus Kinase 1 (JAK1) et la tyrosine kinase 2 (TYK2) sont activées suite à la stimulation des récepteurs de l’IFN I et phosphorylent les facteurs de transcription STAT1 et STAT2, conduisant à l’expression de nombreux ISGs. Les analyses génétiques, de conformation tridimensionnelle, sur un modèle cellulaire in vitro (HEK293T) et ex vivo sur cellules mononuclées périphériques des patients nous ont ainsi permis de mettre en évidence pour ces mutations un caractère constitutionnellement activé, indépendant de la liaison au ligand cGAMP, mais transmettant ce signal à travers la voie d’aval par TBK1. (...) / Type I interferons (IFN I) are antiviral cytokines with potent properties. Hence, the induction, transmission and resolution of the immune response generated by IFN I is tightly regulated. The concept of the type I interferonopathies, recently formulated by our team, rests on the assumption that some diseases arise from a disturbance of this complex signalling pathway, leading to excessive and inappropriate IFN I secretion. On this basis, targeted therapeutics should improve or cure features of such type I interferonopathies, thereby providing a validation of the underlying hypothesis. This PhD project initially focused on the clinical and biological characterisation of monogenic and polygenic interferonopathies, and secondarily on the molecular identification of novel mutations in the gene TMEM173 causing the interferonopathy called STING associated vasculopathy with onset in infancy (SAVI), an auto-inflammatory syndrome with severe cutaneous and pulmonary features. Our selection of patients in comparison to healthy controls was made possible through the use of novel screening tools: IFN signature (qPCR of 6 IFN stimulated genes – ISGs), and measurement of IFN alpha protein levels in serum or plasma (SIMOA-single molecule array - enabling the detection of molecules of IFN in the femtogram [10-18g]) range. In this way, we have been able to expand the phenotypic spectrum of the interferonopathies, which was initially considered as primarily neurological. Patients with Aicardi-Goutières syndrome (AGS), the first described of the monogenic interferonopathies, exhibit dystonia, spasticity, developmental delay, intra-cranial calcifications and white matter abnormalities. However, the systematic use of our interferon screening assays, plus the advent of next-generation sequencing technology, has revealed a much broader set of features relevant to this novel disease grouping – involving the skin (chilblains, necrotising vasculitis, scleroderma), lungs (isolated lung interstitial disease or associated with other signs), musculoskeletal system (joint pain, arthritis, Jaccoud’s arthropathy, muscle pain and myositis), eyes (glaucoma), kidneys (lupus nephritis) and gastro-intestinal tract (early inflammatory bowel disease), as well features of autoimmunity and immunodeficiency. Using our screening assays enabled us to identify three patients variably exhibiting the core features of SAVI, all of whom were found to harbour distinct novel activating mutations in STING. These mutations highlight a protein domain not previously implicated in the control of IFN I signalling. STING is an endoplasmic reticulum protein, acting as a cytosolic adaptor of intracellular sensors of viral DNA in the type I IFN signalling pathway. STING activates TANK-binding kinase (TBK1), allowing transcription of IFN I through phosphorylation of IRF3. Janus kinase 1 (JAK1) and tyrosine kinase 2 (TYK2) are activated following stimulation of the IFN I receptor, leading to phosphorylation of the transcription factors STAT1 and STAT2 and the subsequent induction of a large number of ISGs. Genetic analysis, conformational studies, an in vitro cellular model (HEK293T) and ex vivo experimental data (using patient peripheral blood mononuclear cells - PBMCs) enabled us to confirm the constitutive activating nature of these variants, and show that this activation did not require binding with cGAMP, but was dependent on signalling through TBK1. Ruxolitinib, a JAK1/2 inhibitor, could antagonise this constitutive activation ex vivo. These results indicate a promising therapeutic approach in such patients, and more widely in the monogenic, and perhaps even, polygenic, interferonopathy context.

In order to evaluate the effects of an integrated curriculum on the learning of popular music, the Sting Curriculum was designed for senior secondary students of mixed ability. This nine week program was presented to a sample of 124 students aged between 16 and 18 years in urban Sydney (Australia).The results of tests conducted indicate that students in the sample achieved high scores when a greater emphasis was placed on performance than on the listening and composition activities. The principal findings of the study suggest that the Sting Curriculum was successful as a vehicle for learning popular music, providing students with an integrated and sequential program that motivated participants to become immersed in the music. Furthermore, in the context of an integrated curriculum, popular music learning was enhanced when teachers utilised a pedgogical approach which emphasised the performance activity. / Doctor of Philosophy (PhD)

Polistes dominula and P. fuscatus often nest in bird nest boxes. Potential competition between wasps and birds was studied by removing wasp nests from some boxes. No difference in nesting success of breeding birds was found between boxes with wasp nests and those in which wasp nests were removed. Boxes that never had a wasp nest and boxes from which wasp nests were removed differed greatly in bird occupancy, suggesting that birds detected previous wasp presence. Some bird species gain protection by nesting near wasp nests. Birds may prefer to nest near wasp species that inflict higher sting pain levels. A rank correlation of data from published studies provided no evidence that pain level influences which wasp nests are most attractive to nesting birds. A comprehensive table of bird-wasp nesting associations (listing 121 bird species, 28 wasp species and 4 bee species) is included here.

Nesta dissertação pretendo reler o debate entre Ronald Dworkin e o positivismo jurídico. Farei isto sob o prisma da filosofia analítica, especificamente, contextualizando o debate no âmbito de uma discussão travada entre três teorias semânticas específicas: a descricional, o externalismo semântico e o bi-dimensionalismo ambicioso. Há algum tempo Dworkin lançou uma crítica ao positivismo, qual seja, o positivismo jurídico pretende reduzir a forma direito de como as coisas são à conformação puramente descritiva de como o mundo é. Disse, ainda, que somente quando concebido como uma teoria semântica é que o positivismo jurídico tornar-se-ia inteligível. Os posivistas, a seu turno, argumentam que a Jurisprudência analítica é um projeto teórico pelo direito e não pelo significado do termo direito e que, portanto, deveríamos manter separados dois tipos de questionamentos: O que é direito? e O que é direito?. Se tudo correr bem, ao reler o debate entre os positivistas e Ronald Dworkin a partir do instrumental obtido no âmbito da teoria semântica, poderemos perceber que pode ser verdade que o positivismo jurídico, enquanto projeto teórico, é sobre o direito, o referente, e não sobre o direito, o termo; entretanto, a forma pela qual o positivismo concebe o questionamento O que é direito?, ele mesmo, parece acabar por qualificá-lo, num sentido não trivial, como semântico. / I intend to reread the debate between Ronald Dworkin and legal positivism. I will do that through the prism of analytic philosophy, specifically in the context of the debate between three specific semantic theories: descriptional, externalism and the ambitious bidimensionalism. Dworkin criticized legal positivism: the legal positivism aims to reduce the law-way of things to the purely descriptive form of the world. He also said that only when conceived as a semantic theory is that legal positivism would become intelligible. The posivists argue that analytical Jurisprudence is a theoretical project about law and not about the meaning of \"law\", therefore we should keep separated two types of questions: \"What is law?\" and \"What is \'law\'?\". If all goes well, when rereading the debate through the prism of the discussion in the context of semantic theories, we will realize that it may be true that legal positivism is about law, the referent, and not about \"law\". However, the way in which positivism conceives the question \"What is law?\" seems to qualify it as semantic in a nontrivial sense.

This thesis was funded by the Natural Environment Research Council (NERC) and is presented in an alternative thesis format. The thesis consists of three separate journal articles which form a coherent research project. Paper 1 investigates the development of strong winds in a dry, idealised extratropical cyclone using the horizontal momentum equation. In particular, the southwest wind maximum that develops was found to contain air parcels from three airstreams. The development of the horizontal along-flow forces around the cyclone and along trajectories entering the southwest wind maximum were analysed. An attempt to extend this methodology to a moist, idealised extratropical cyclone was made. However, the effect of adding moisture to the initial condition was found to be negligible. The reasons for this are explored in Paper 2, which documents this finding: that the effect of moisture on the development of an idealised, baroclinic wave is sensitive to the choice of initial condition. Paper 3 applies the horizontal momentum equation diagnostics to an intense, marine extratropical cyclone that brought strong winds to Ireland and the United Kingdom on 12 February 2014. The development of strong winds in Cyclone Tini was investigated by turning off latent heat release and surface fluxes. In the absence of latent heat release a weaker wind maximum developed. However, the simulation without surface fluxes had a very similar vertical structure of the horizontal wind to the full-physics simulation, but a weaker surface wind maximum. The reason for this weaker wind maximum was analysed using the quasigeostrophic omega equation. This analysis demonstrated a maximum in forcing for descent southwest of the low both in the full-physics simulation and in the simulation without surface fluxes, however strong winds were prevented from reaching the surface in the simulation without surface fluxes because of a more stable boundary layer around the bent-back front.