La biologie moléculaire et, plus spécifiquement, la régulation de l’expression génique ont été révolutionnées par la découverte des microARN (miARN). Ces petits ARN d’une vingtaine de nucléotides sont impliqués dans la majorité des processus cellulaires et leur expression est dérégulée dans plusieurs maladies, comme le cancer. Un miARN reconnaît ses cibles principalement par son noyau, ce qui lui permet de réguler simultanément la traduction de centaines d’ARN messagers. Nos travaux ont montré l’existence d’une boucle de rétro-activation négative, entre deux miARN du polycistron miR-17-92 et trois facteurs de transcription de la famille E2F. E2F1, 2 et 3 induisent la transcription de miR-20 et miR-17 qui par la suite inhibent leur traduction. Nos résultats suggèrent l’implication de cette boucle dans la résistance à l’apoptose induite par E2F1 dans les cellules du cancer de la prostate, ce qui expliquerait en partie le potentiel oncogénique du polycistron miR-17-92. L’étude de ce motif de régulation nous a donc permis de réaliser le potentiel incroyable qu’ont les miARN à inhiber la traduction de plusieurs gènes. Basé sur les règles de reconnaissance des miARN, nous avons développé et validé MultiTar. Cet outil bioinformatique permet de trouver la séquence d’un miARN artificiel ayant le potentiel d’inhiber la traduction de gènes d’intérêts choisis par l’utilisateur. Afin de valider MultiTar, nous avons généré des multitargets pouvant inhiber l’expression des trois E2F, ce qui nous a permis de comparer leur efficacité à celle de miR-20. Nos miARN artificiels ont la capacité d’inhiber la traduction des E2F et de neutraliser leur fonction redondante de la progression du cycle cellulaire de façon similaire ou supérieur à miR-20. La fonctionnalité de notre programme, ouvre la voie à une stratégie flexible pouvant cibler le caractère multigénique de différents processus cellulaires ou maladies complexes, tel que le cancer. L’utilisation de miARN artificiels pourrait donc représenter une alternative intéressante aux stratégies déjà existantes, qui sont limitées à inhiber des cibles uniques. En plus d’élucider un réseau de régulation complexe impliquant les miARN, nous avons pu tirer profit de leur potentiel d’inhibition par la conception de miARN artificiels. / miRNAs are powerful regulators of gene expression in mammals. These small RNAs of around 20 nucleotides are involved in several cellular processes and diseases. MiRNAs recognize their targets mainly by a region comprising nucleotides 2-8, known as the seed. This characteristic gives them the potential to inhibit hundreds of messenger RNAs. Our first goal was to better characterize the complex network involving miRNAs in the regulation of gene expression. To achieve this, we studied the relation between a family of transcription factors, the E2Fs, and a family of miRNAs, the miR-17-92 cluster. Our results suggest a negative feedback loop involving miR-17, miR-20a, E2F1, E2F2 and E2F3. In this loop E2F1, 2 and 3 activate the transcription of the two miRNAs that inhibit their translation in return. The inhibition of the antiapoptotic function of E2F1 by miR-17 and miR-20 in a prostate cancer context, could explain the oncogenic potential of the miR-17-92 cluster that was previously reported. Studying the miR-20/E2F feedback loop made us realize how powerful was the ability of miRNAs to inhibit several targets. To overcome the lack of efficient tools able to inhibit simultaneously the expression of multiple genes, our second goal was to develop MultiTar, an algorithm able to design artificial miRNAs that target a set of predetermined genes. MultiTar was validated in silico, using known targets of endogenous miRNAs and in vivo, taking advantage of our experience with the E2F context. We designed artificial miRNAs against E2F1-3 and expressed them both in normal human fibroblasts and prostate cancer cells where they inhibited cell proliferation and induced cellular senescence. The observed phenotypes were precisely those known for inhibiting E2F activities. Hence, MultiTar can efficiently design artificial micro RNAs able to target multiple genes and is thus a flexible tool that can address the issue of multigenic diseases and complex cellular processes. The use of multitargets could be an alternative to overcome the limits of drugs or siRNAs that are designed generally to regulate only one target.

This work builds up on the greatest minds of Cambridge Holography: Adrian Cable, Edward Buckley, Jonathan Freeman, and Christoph Bay. Cable and Buckley, developed an OSPR algorithm which was the first to provide high-quality real-time hologram generation using general-purpose hardware while Freeman designed a method to correct arbitrary aberrations. As ingenious as the method was, the calculations were extensively lengthy. Addressing this issue, a variant of OSPR suited for correcting spatially-varying aberration is presented. The algorithm combines the approaches of Cable, Buckley and Freeman to provide real-time hologram generation while incorporating various corrections (aberration, distortion, and pixel shape envelope). A high-performance implementation on a mid-range GPU achieved hologram generation up to 12 fps. Following topic studied is an adaptive optical correction. This work attempts to construct a set of methods, forming an automated testbed for holographic projectors. Each model, after exiting the production line is placed on such testbed, having all of its imperfections characterized. Once calibrated, each model is able to display highest-quality image throughout its life-span. An application of this work to industry was carried in collaboration with Dr Phillip Hands (University of Edinburgh) and LumeJET. Three demonstrators are constructed intending for a cost-effective system for holographic lithography. They are characterized using the developed testbed. Using the supersampled Adaptive OSPR algorithm, the diffraction limit was surpassed 2.75 times allowing to increase the patterning area. This combines approaches of Cable, Buckley, Freeman and Bay to achieve a wide field-of-view and high pixel-count replay field using off-the-shelf components. This thesis is finished describing the work on 3D holography carried with Penteract28. It is shown that the 2D hologram in the presence of spatially-varying aberrations is mathematically equivalent to a 3D hologram. The same implementation of the algorithm can be used to provide real-time 3D hologram generation.

Self-adaptive systems are capable of autonomously adjusting their behavior at runtime to accomplish particular adaptation goals. The most common way to realize self-adaption is using a feedback loop(s) which contains four actions: collect runtime data from the system and its environment, analyze the collected data, decide if an adaptation plan is required, and act according to the adaptation plan for achieving the adaptation goals. Existing approaches achieve the adaptation goals by using formal methods, and exhaustively verify all the available adaptation options, i.e., adaptation space. However, verifying the entire adaptation space is often not feasible since it requires time and resources. In this thesis, we present an approach which uses machine learning to reduce the adaptation space in self-adaptive systems. The approach integrates with the feedback loop and selects a subset of the adaptation options that are valid in the current situation. The approach is applied on the simulator of a self-adaptive Internet of Things application which is deployed in KU Leuven, Belgium. We compare our results with a formal model based self-adaptation approach called ActivFORMS. The results show that on average the adaptation space is reduced by 81.2% and the adaptation time by 85% compared to ActivFORMS while achieving the same quality guarantees.

No description available.

Aufgrund des weltweit steigenden Energiebedarfs, dessen Deckung derzeit größtenteils auf fossilen Brennstoffen basiert, ist es nötig geworden, die Entwicklung alternativer Möglichkeiten zur Erzeugung von Elektroenergie als Primärenergie voranzutreiben. Eine dieser alternativen Möglichkeiten ist die Brennstoffezellentechnologie, welche sowohl in stationären als auch mobilen Anwendungen zum Einsatz kommen kann. Ihrer weitreichenden Verbreitung stehen bislang die aufgrund des großen Fertigungsaufwandes hohen Herstellungskosten der benötigten Komponenten im Wege. Hierzu zählen die Gasdiffusionslagen des weit verbreiteten Typs der wasserstoffbetriebenen Polymerelektrolytbrennstoffzelle. Es treten zwischen den einzelnen Fertigungsschritten im Herstellungsprozess dieser Gasdiffusionslagen Wechselwirkungen auf, die zu unerwünschten Materialveränderungen führen. Die Ursachen dieser Wechselwirkungen sind nicht vollends verstanden. Eine Vertiefung des Verständnisses der Herstellungsprozesse soll die Grundlage für eine Optimierung der Prozessführung bilden. Es sollen eine Kostenreduktion sowie eine Leistungssteigerung der Gasdiffusionslagen ermöglicht werden.:1 Einleitung 1 2 Stand der Technik 5 2.1 Brennstoffzellen 5 2.2 Gasdiffusionslagen 11 3 Problemstellung und Zielsetzung 17 4 Analyse und Klassifizierung von GDL-Fehlern 20 4.1 Fehlerklassifizierung 22 4.2 Fehleridentifizierung 26 4.3 Auswahl zu analysierender Fehlerbilder 27 4.4 Charakteristika der ausgewählten Fehlerbilder 42 4.4.1 Bahndeformationen 42 4.4.2 Umlaufende Verdickungen von Wickeln in Umfangsrichtung 44 4.4.3 Längs- und Queraufrauhungen sowie Rauhspuren 45 5 Theoretische Grundlagen 49 5.1 Physikalische und mechanische Grundlagen 49 5.1.1 Zug-, Biege- und Druckspannungen in Warenbahnen 49 5.1.2 Elastizitäts- und Kompressionsmoduli 52 5.1.3 Elastizität und Plastizität 53 5.1.4 Umformmechanismen im GDL-Basisvliesstoff und Versagensarten von Fasern 54 5.2 Statistik 55 5.2.1 Korrelationsanalyse 55 5.2.2 Regressionsanalyse 56 5.2.3 Zweistichproben-t-Tests und Konfidenzintervalle 56 5.2.4 Stichprobenumfang 57 5.3 Qualitätsregelkreise 58 6 Eingesetzte Untersuchungsmethoden 60 6.1 Mechanische Eigenschaften 64 6.1.1 Höchstzugkraft und Höchstzugkraftdehnung 64 6.1.2 Elastizitätsmodul und Kompressibilität 66 6.1.3 Elastische und plastische Deformation bei Zugbelastungen 67 6.1.4 Flächenmasse 70 6.1.5 Biegesteifigkeit 72 6.1.6 Dickenmessung 74 6.2 Thermische Eigenschaften 75 6.2.1 Wärmeleitfähigkeit 75 6.3 Bildgebende Verfahren 78 6.3.1 Schliffbildmikroskopie 78 6.3.2 Rasterelektronenmikroskopie 78 6.3.3 µ-Computertomographie 79 7 Herstellungsverfahren der untersuchten Gasdiffusionslagen im Überblick 81 8 Basisvliesstoffherstellung 84 8.1 Prozess der Vliesbildung und Verfestigung 84 8.2 Charakterisierung des GDL-Basisvliesstoffes 90 8.3 Fehlerbilder des GDL-Basisvliesstoffes 103 9 Dickenkalibrierung 113 9.1 Prozess der Dickenkalibrierung des GDL-Basisvliesstoffes 113 9.2 Charakterisierung des dickenkalibrierten GDL-Basisvliesstoffes 120 9.3 Fehlerbilder des dickenkalibrierten GDL-Basisvliesstoffes 130 9.3.1 Prozessbeobachtung 130 9.3.2 Hypothesenbildung und Verifikation 135 9.3.3 Maßnahmen zur Fehlervermeidung 146 10 Carbonisierung 156 10.1 Prozess der Carbonisierung 156 10.2 Charakterisierung carbonisierten GDL-Substrates 157 10.3 Fehlerbilder im Carbonisierprozess 163 11 Data Mining für die GDL-Herstellung 167 11.1 Datenerhebung 167 11.2 Auszuwertende Parameter 172 11.3 Ergebnisse der Parameteranalysen 173 12 Qualitätsregelkreise zum GDL-Produktionsprozess 178 12.1 Wulstbildung und Längsaufrauhung 178 12.2 Queraufrauhung 181 13 Zusammenfassung und Ausblick 184 14 Literaturverzeichnis 186 15 Abbildungsverzeichnis 192 16 Abkürzungsverzeichnis 201 17 Formelverzeichnis 203 18 Anlagenverzeichnis 204 / Due to worldwide increasing energy consumption, which is mainly covered by fossile fuels nowadays, it has become a necessity to further develop alternative possibilities to create electricity as primary energy. One alternative technology to accomplish this is fuel cell technology which can be used in stationary as well as in mobile applications. One aspect hindering its widespread use is the high manufacturing cost of the needed components due to the complicated production processes. Among these are gad diffusion layers of the commonly used hydrogen-driven polymer electrolyte fuel cells. There are interactions occurring between the several production steps leading to unwanted changes in material properties. The causes of these interactions are not completely understood. A deeper understanding of these shall be the basis for optimizations in process design and therefore cost reductions and improvements in performance of gas diffusion layers can be achieved.:1 Einleitung 1 2 Stand der Technik 5 2.1 Brennstoffzellen 5 2.2 Gasdiffusionslagen 11 3 Problemstellung und Zielsetzung 17 4 Analyse und Klassifizierung von GDL-Fehlern 20 4.1 Fehlerklassifizierung 22 4.2 Fehleridentifizierung 26 4.3 Auswahl zu analysierender Fehlerbilder 27 4.4 Charakteristika der ausgewählten Fehlerbilder 42 4.4.1 Bahndeformationen 42 4.4.2 Umlaufende Verdickungen von Wickeln in Umfangsrichtung 44 4.4.3 Längs- und Queraufrauhungen sowie Rauhspuren 45 5 Theoretische Grundlagen 49 5.1 Physikalische und mechanische Grundlagen 49 5.1.1 Zug-, Biege- und Druckspannungen in Warenbahnen 49 5.1.2 Elastizitäts- und Kompressionsmoduli 52 5.1.3 Elastizität und Plastizität 53 5.1.4 Umformmechanismen im GDL-Basisvliesstoff und Versagensarten von Fasern 54 5.2 Statistik 55 5.2.1 Korrelationsanalyse 55 5.2.2 Regressionsanalyse 56 5.2.3 Zweistichproben-t-Tests und Konfidenzintervalle 56 5.2.4 Stichprobenumfang 57 5.3 Qualitätsregelkreise 58 6 Eingesetzte Untersuchungsmethoden 60 6.1 Mechanische Eigenschaften 64 6.1.1 Höchstzugkraft und Höchstzugkraftdehnung 64 6.1.2 Elastizitätsmodul und Kompressibilität 66 6.1.3 Elastische und plastische Deformation bei Zugbelastungen 67 6.1.4 Flächenmasse 70 6.1.5 Biegesteifigkeit 72 6.1.6 Dickenmessung 74 6.2 Thermische Eigenschaften 75 6.2.1 Wärmeleitfähigkeit 75 6.3 Bildgebende Verfahren 78 6.3.1 Schliffbildmikroskopie 78 6.3.2 Rasterelektronenmikroskopie 78 6.3.3 µ-Computertomographie 79 7 Herstellungsverfahren der untersuchten Gasdiffusionslagen im Überblick 81 8 Basisvliesstoffherstellung 84 8.1 Prozess der Vliesbildung und Verfestigung 84 8.2 Charakterisierung des GDL-Basisvliesstoffes 90 8.3 Fehlerbilder des GDL-Basisvliesstoffes 103 9 Dickenkalibrierung 113 9.1 Prozess der Dickenkalibrierung des GDL-Basisvliesstoffes 113 9.2 Charakterisierung des dickenkalibrierten GDL-Basisvliesstoffes 120 9.3 Fehlerbilder des dickenkalibrierten GDL-Basisvliesstoffes 130 9.3.1 Prozessbeobachtung 130 9.3.2 Hypothesenbildung und Verifikation 135 9.3.3 Maßnahmen zur Fehlervermeidung 146 10 Carbonisierung 156 10.1 Prozess der Carbonisierung 156 10.2 Charakterisierung carbonisierten GDL-Substrates 157 10.3 Fehlerbilder im Carbonisierprozess 163 11 Data Mining für die GDL-Herstellung 167 11.1 Datenerhebung 167 11.2 Auszuwertende Parameter 172 11.3 Ergebnisse der Parameteranalysen 173 12 Qualitätsregelkreise zum GDL-Produktionsprozess 178 12.1 Wulstbildung und Längsaufrauhung 178 12.2 Queraufrauhung 181 13 Zusammenfassung und Ausblick 184 14 Literaturverzeichnis 186 15 Abbildungsverzeichnis 192 16 Abkürzungsverzeichnis 201 17 Formelverzeichnis 203 18 Anlagenverzeichnis 204

A slab-coupled optical fiber sensor (SCOS) falls into a narrow class of all-dielectric optical fiber electric field sensors, which makes it a perfect candidate for measurements of high electric fields in environments where presence of conductors is highly perturbing to the system under test. Its nonlinear response to high fields requires a new nonlinear calibration technique. A nonlinear calibration method is explained and demonstrated to successfully measure high electric fields, as well as high voltages with dynamic range up to 50 dB. Furthermore, a SCOS can be fitted into narrow spaces and make highly localized measurements due to its small size. This allows a SCOS to be integrated inside a standard high voltage coaxial cable, such as RG-218. Effects of partial discharge and arcing is minimized by development of a fabrication method to avoid introduction of impurities, especially air-bubbles, into the cable during SCOS insertion. Low perturbation of the measured voltage is shown by simulating the introduced voltage reflections to be on the order of –50 dB. It is also shown that a SCOS can be inserted into other cables without significant perturbation to the voltage. A complete high voltage and high electric field measurement system is built based on the high-voltage modifications of the SCOS technology. The coaxial SCOS is enhanced for robustness. Enhancements include packaging a SCOS into stronger ceramic trough, strengthening the fiber with kevlar reinforced furcation tubing and protecting the sensor with metal braces and protective shells. The interrogator is protected from electromagnetic interference with an RF-shielded box. Reduction in power losses introduced by the new PANDA-SCOS technology allows interrogator bandwidths to be increased up to 1.2 GHz. The whole measurement process is streamlined with dedicated software, developed specifically for high voltage and electric field measurements with support for the nonlinear calibration.

A slab-coupled optical fiber sensor (SCOS) falls into a narrow class of all-dielectric optical fiber electric field sensors, which makes it a perfect candidate for measurements of high electric fields in environments where presence of conductors is highly perturbing to the system under test. Its nonlinear response to high fields requires a new nonlinear calibration technique. A nonlinear calibration method is explained and demonstrated to successfully measure high electric fields, as well as high voltages with dynamic range up to 50 dB. Furthermore, a SCOS can be fitted into narrow spaces and make highly localized measurements due to its small size. This allows a SCOS to be integrated inside a standard high voltage coaxial cable, such as RG-218. Effects of partial discharge and arcing is minimized by development of a fabrication method to avoid introduction of impurities, especially air-bubbles, into the cable during SCOS insertion. Low perturbation of the measured voltage is shown by simulating the introduced voltage reflections to be on the order of −50 dB. It is also shown that a SCOS can be inserted into other cables without significant perturbation to the voltage.A complete high voltage and high electric field measurement system is built based on the high-voltage modifications of the SCOS technology. The coaxial SCOS is enhanced for robustness. Enhancements include packaging a SCOS into stronger ceramic trough, strengthening the fiber with kevlar reinforced furcation tubing and protecting the sensor with metal braces and protective shells. The interrogator is protected from electromagnetic interference with an RF-shielded box. Reduction in power losses introduced by the new PANDA-SCOS technology allows interrogator bandwidths to be increased up to 1.2 GHz. The whole measurement process is streamlined with dedicated software, developed specifically for high voltage and electric field measurements with support for the nonlinear calibration.

As an object at the boundary between virtual and physical reality, the screen exists both as a displayer and as a thing displayed, thus functioning as a mediator. The screen's virtual imagery produces a sense of immersion in its viewer, yet at the same time the materiality of the screen produces a sense of rejection from the viewer's complete involvement in the virtual world. The experience of the screen is thus an oscillation between these two states of immersion and rejection. Nowadays, as interactivity becomes a central component of the relationship between viewers and many artworks, the viewer experience of the screen is changing. Unlike the screen experience in non-interactive artworks, such as the traditional static screen of painting or the moving screen of video art in the 1970s, interactive media screen experiences can provide viewers with a more immersive, immediate, and therefore, more intense experience. For example, many digital media artworks provide an interactive experience for viewers by capturing their face or body though real-time computer vision techniques. In this situation, as the camera and the monitor in the artwork encapsulate the interactor's body in an instant feedback loop, the interactor becomes a part of the interface mechanism and responds to the artwork as the system leads or even provokes them. This thesis claims that this kind of direct mirroring in interactive screen-based media artworks does not allow the viewer the critical distance or time needed for self-reflection. The thesis examines the previous aesthetics of spatial and temporal perception, such as presentness and instantaneousness, and the notions of passage and of psychological perception such as reflection, reflexiveness and auratic experience, looking at how these aesthetics can be integrated into new media screen experiences. Based on this theoretical research, the thesis claims that interactive screen spaces can act as a site for expression and representation, both through a doubling effect between the physical and virtual worlds, and through manifold spatial and temporal mappings with the screen experience. These claims are further supported through exploration of screen-based media installations created by the author since 2003.

Made available in DSpace on 2015-04-20T12:33:26Z (GMT). No. of bitstreams: 1 Leandro.pdf: 2831865 bytes, checksum: 5804d85c95f338cf4054c799f4dfd45d (MD5) Previous issue date: 2011-10-07 / Voice calls based on Voice over Internet Protocol (VoIP) technology are liable to several impairments from both application and network layer, such as codec compression, end-to-end delay, and packet loss. For years, this problem has been challenging researchers and practitioners, who have been designing and improving QoS control mechanisms for VoIP applications. Such mechanisms aim to make optimum use of network and terminal resources so as to minimize the effects of network impairments on voice quality. Among the several proposed QoS control mechanisms for VoIP, some of them seek to adapt the voice flow or other VoIP-related parameters in accordance with significant changes in the network, end users preferences, or service providers requirements. VoIP systems are particularly likely to require a dynamic adaptation solution for dealing with the complex trade-off between speech quality and impairments, because of the decentralized control nature of IP networks and the stochastic nature of data packet delivery. Although the existing adaptive solutions for QoS control of VoIP show some performance improvement and exhibit some sort of feedback, they do not provide explicit focus on the control loop. This document shows the current progress of our thesis, which addresses the adjustment of internal parameters of VoIP terminals (at application layer) that affect the voice flow, with the aim of improving speech quality in response to changes in network conditions. It is not in the scope of the thesis to propose adaptive solutions that focus exclusively on signaling, billing, security issues, or operate at the network layer. Therefore, this thesis addresses the problem of how adjust encoding parameters in response to variations in delay and packet loss, in order to optimize speech quality. The objective is to optimize user-perceptible attributes of speech, under the perspective of self-adaptive software systems. The emphasis is not to develop new audio codecs, but to build a control loop in the core of sender and receiver terminals to adapt voice flow settings according to network conditions. The main contributions of this thesis are the following: determination of user s perception during codec switching; parametrization of codec precedence for supporting codec switching decision; explicit design of a monitoring analysis planning execution control loop as the core of the adaptation process; and efficiency analysis of feedback message exchanging. / Chamadas de voz baseadas na tecnologia VoIP (Voice over Internet Protocol) estão suscetíveis a degradações diversas, provenientes tanto da camada de aplicação, como da camada de rede, tais como compressão do codec, atraso fim a fim e perda de pacotes. Durante anos, esse problema tem desafiado pesquisadores e profissionais, que têm concebido e melhorado mecanismos de controle de QoS para aplicações VoIP. Tais mecanismos visam otimizar a utilização dos recursos da rede e do terminal VoIP de modo a minimizar os efeitos deletérios da rede subjacente sobre a qualidade de voz. Entre as várias propostas de mecanismos de controle de QoS para VoIP, alguns deles procuram adaptar o fluxo de voz ou outros parâmetros VoIP de acordo com mudanças significativas na rede, preferências de usuário, ou requisitos dos provedores de serviços VoIP. Sistemas VoIP particularmente exigem soluções de adaptação dinâmica para lidar com a complexa relação de compromisso entre qualidade de voz e fatores de degradação, por causa da natureza descentralizada e estocástica das redes IP na entrega de pacotes de voz. Embora as soluções adaptativas existentes para controle de QoS em VoIP mostrem alguma melhora de desempenho e apresentem algum tipo de feedback, elas não fornecem foco explícito na ciclo de controle (control loop). Este documento mostra o progresso atual da nossa tese, que aborda o ajuste de parâmetros internos de terminais VoIP (camada de aplicação) que afetam o fluxo de voz, com o objetivo de melhorar a qualidade da fala em resposta a mudanças nas condições da rede. Não faz parte do escopo da tese abordar soluções adaptativas que se concentram exclusivamente em sinalização, bilhetagem, problemas de segurança, ou que operam no nível da camada de rede. Portanto, esta tese aborda o problema da concepção e avaliação de estratégias adaptativas que explorem as relações de compromisso entre qualidade da fala e os seguintes fatores de degradação: compressão do codec, atraso fim a fim e perda de pacotes. A finalidade é otimizar atributos da fala perceptíveis aos usuário, sob a perspectiva de sistemas de software autoadaptativo. A ênfase não reside em desenvolver novos codecs de áudio, mas sim em desenvolver um ciclo de controle como entidade central de um terminal VoIP, que possa adaptar as configurações do fluxo de voz de acordo com as condições da rede. As principais contribuições desta tese são as seguintes: determinação da percepção do usuário durante a comutação de codec; parametrização de precedência de codecs para suporte de decisão de comutação de codec; enfoque no ciclo de controle baseado nas atividades de monitoramento análise planejamento execução como núcleo do processo de adaptação; e análise de eficiência de troca de mensagens de feedback.

Le cancer papillaire de la thyroïde (CPT) est la tumeur endocrine la plus fréquente. Des anomalies moléculaires activant la voie des MAPK (Mitogen-Activated Protein Kinases) sont identifiées, de façon mutuellement exclusive, dans environ 70% des cas. Il s’agit de réarrangements chromosomiques, le plus souvent de type RET/PTC (10%), de mutations ponctuelles activatrices des trois isoformes de l’oncogène RAS (H, N et K-RAS) (10%), ou de l’oncogène B-RAF (50%). La mutation « hot spot » B-RAFV600E est la plus fréquemment identifiée, elle est associée à une plus grande agressivité clinique (diagnostic à un stade tardif, risque de récidives et de décès accru). Ces évènements moléculaires ont pour conséquence commune l’activation de la voie des MAPK, se traduisant en aval par la phosphorylation de MEK (Mitogen-activated Extracellular signal-Regulated Kinase) puis de ERK (Extracellular signal-Regulated Kinase). Cette dernière est régulée négativement par des phosphatases, appartenant à la famille des Dual Specificity Phosphatases (DUSPs), d’expression ubiquitaire, et en particulier de deux phosphatases spécifiques de ERK, l’une cytoplasmique (DUSP6) et l’autre nucléaire (DUSP5). Nous avons fait l’hypothèse que ces phosphatases pouvaient être soit des gènes suppresseurs de tumeurs (leur perte d’expression conduisant à une augmentation de phosphorylation de ERK et une prolifération accrue), soit des marqueurs du degré d’activation de la voie MAPK dans le cadre d’une boucle de rétrocontrôle négatif. Ceci nous a conduits à analyser la régulation et l’expression de ces phosphatases dans trois modèles : la lignée cellulaire PCCL3 (thyroïde de rat), exprimant l’un des trois principaux oncogènes mutés dans les CPT (RET/PTC3 ou H-RASV12 ou B-RAFV600E) sous le contrôle d’un promoteur inductible par la doxycycline, des lignées cellulaires humaines dérivant de CPT et des CPT humains. (...) / Papillary thyroid cancer (PTC) is the most common endocrine malignancy. Mutually exclusive and activating alterations of the MAPK pathway (Mitogen-Activated Protein Kinases) are identified in 70% of cases. Common mutations found in PTCs are point mutation of the B-RAF (50%) and RAS genes (10%) as well as RET/PTC chromosomal rearrangements (10%). The hot spot B-RAFV600E mutation is the most frequently alteration identified and is connected with agressive clinical characteristics (high stage at diagnosis, high recurrence risk and death). These molecular events lead to constitutive activation of the MAPK pathway, resulting in MEK (Mitogen-activated Extracellular signal-Regulated Kinase) and ERK (Extracellular signal-Regulated Kinase) phosphorylation. ERK is negatively regulated by phosphatases and among them, Dual Specificity Phosphatases (DUSPs), ubiquitary expressed, in particular two ERK-specific phosphatases DUSP5 (nuclear) and DUSP6 (cytosolic). We hypothesized that these phosphatases could have tumor supressor properties (i.e. their loss would be associated with an increase in MAPK pathway activation) or may serve as a surrogate marker of MAPK pathway activation in the context of a negative feedback loop. We analysed regulation and expression of both phosphatases in 3 models: three PCCL3 cell lines (rat thyroid cells) expressing one of the most common oncogene identified in PTCs (RET/PTC3 or H-RASV12 or B-RAFV600E) under the control of a doxycycline-inducible promoter, human PTC-derived cell lines and human PTC. We demonstrated that MAPK pathway activation was correlated with induction of DUSP5 and DUSP6. These phosphatases are involved in a negative feedback loop that contributes to a tight regulation of phospho-ERK levels. DUSP5 and DUSP6 mRNA are overexpressed in human PTCs, especially in B-RAF mutated tumors suggesting a higher MAPK signaling output in these agressive PTCs. Silencing of DUSP5 and/or DUSP6 by small interfering RNA does not affect proliferation of human B-RAFV600E thyroid carcinoma-derived cell lines, suggesting the lack of tumor suppressor gene role. Compensatory changes in expression of DUSPs when a specific one is inactivated may explain this lack of effect. On the opposite, a DUSP6 pharmacological inhibitor induced a concentration dependent decrease in proliferation of human B-RAFV600E cells, suggesting « off-target » effect of this inhibitor. In a second part, we analysed the regulation of DUSP5 expression, which is a target of the MAPK pathway activation. We demonstrated, using pharmacological inhibitors, that DUSP5 is an early response gene, regulated mostly by the MAPK pathway, at the transcriptional level. Two contiguous CArG boxes that bind serum response factor (SRF) were found in a 1Kb promoter region, as well as several E twenty-six transcription factor family binding sites (EBS). These sites potentially bind Elk-1, a transcription factor activated by ERK1/2. Using wild type or mutated DUSP5 promoter reporters, we demonstrated that SRF plays a crucial role in serum induction of DUSP5 promoter activity, the proximal CArG box being important for SRF binding in vitro and in living cells. Moreover Elk-1 was bound in vitro to a promoter region containing the proximal CArG box and a putative EBS. Its specific binding to SRF was necessary to elicit promoter response to dominant positive Elk-VP16 and to enhance the response to serum stimulation. Altogether our results suggest that the MAPK pathway is more active in B-RAFV600E PTC than in PTC with other genetic alteration and could explain their clinical agressivity. DUSP5 and DUSP6, as well as phosphorylated MEK, are markers of activation of the MAPK pathway. Neither phosphatase has tumor suppressor properties in our thyroid cancer cell models. Our results suggest redundancy and functional compensation among DUSPs. 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