Le système immunitaire inné repose sur la détection de motifs microbiens et l'activation de réponses adaptées, parmi lesquelles les voies de signalisation dépendantes des facteurs NF-κB jouent un rôle primordial. Ces voies sont finement régulées afin d'éviter une réponse immunitaire excessive et soutenue dans le temps qui peut causer de nombreuses pathologies, comme les maladies auto-immunes et pro-inflammatoires. Au cours de ma thèse, j'ai élucidé certains mécanismes de régulation des voies de signalisation NF-κB, Toll et IMD, chez la drosophile, qui reposent sur l'ubiquitination de protéines et leur dégradation par la voie endocytaire ou le protéasome. L'ubiquitination réversible des protéines est une modification post-traductionnelle qui permet de réguler leur activité, leur stabilité et leur localisation subcellulaire. En particulier, l'ubiquitination des récepteurs membranaires peut servir de signal d'endocytose et de dégradation lysosomale. Chez la drosophile, le récepteur PGRP-LC reconnaît spécifiquement le peptidoglycane (PGN) bactérien de type acide diaminopimélique et induit la voie de signalisation IMD. J'ai montré que PGRP-LC est ubiquitiné, internalisé et dégradé par la voie endocytaire. Dans ce processus, j'ai identifié le rôle majeur de la déubiquitinase USP8 qui contrôle la dégradation de PGRP-LC ubiquitiné. J'ai aussi mis en évidence que la stimulation de la voie IMD par les PGN augmente l'internalisation et la dégradation de PGRP-LC, assurant l'élimination des récepteurs après que la voie IMD ait été activée. En outre, j'ai participé à des études visant à comprendre le rôle des déubiquitinases USP2, USP34 et USP36, préalablement sélectionnées par l'équipe comme des régulateurs négatifs des voies IMD et/ou Toll. Mes résultats ont notamment contribué à montrer que USP2 agit principalement au niveau de la protéine adaptatrice Imd, en permettant l'hydrolyse de ses chaînes d'ubiquitine K48 et sa dégradation par le protéasome. Finalement, j'ai observé que USP2 interagit également avec PGRP-LC et favorise l'hydrolyse des chaînes K48 associées à ce récepteur, bien que dans ce cas, la dégradation des formes poly-ubiquitinées K48 de PGRP-LC ne dépende pas du protéasome, mais des protéines de la voie endocytaire Hrs, Rab5 et de la déubiquitinase USP8. / The innate immune system relies on the recognition of “non-self” and on the activation of adapted responses, among which NF-κB signaling pathways play a crucial role. These pathways are tightly regulated, in order to prevent an excessive and sustained immune response, responsible for several pathologies, such as autoimmune and pro-inflammatory diseases. During my PhD thesis, I elucidated some Drosophila regulatory mechanisms of NF-κB pathways, Toll and IMD, which rely on protein ubiquitination and their subsequent degradation by the endocytic pathway or proteasome. Reversible ubiquitination of proteins is a post-translational modification, regulating their activity, their stability and the subcellular localization. In particular, ubiquitination of membrane receptors could trigger their internalization and their subsequent lysosomal degradation. In Drosophila, the PGRP-LC receptor specifically recognizes diaminopimelic acid containing peptidoglycan (PGN) and induces the IMD signaling pathway. I proved that PGRP-LC receptor is ubiquitinated, internalized and degraded by the endocytic pathway. In this process, I identified the major role of the USP8 deubiquitinating enzyme, which controls the degradation of ubiquitinated PGRP-LC. Besides, I showed that the IMD stimulation by PGN enhances the PGRP-LC internalization and its degradation, ensuring receptors elimination once the IMD pathway has been activated. Moreover, I took part to studies, aiming to understand the role of USP2, USP34 and USP36, previously selected by the team as negative regulators of the IMD and/or Toll pathways. In particular, my results showed that USP2 principally acts at the Imd level, allowing for the hydrolysis of its K48 poly-ubiquitin chains and its proteasomal degradation. Finally, I observed that USP2 also interacts with PGRP-LC and favors the hydrolysis of PGRP-LC associated K48 chains, whereas the degradation of K48 poly-ubiquitinated PGRP-LC is independent from the proteasome, but rather depends on the Hrs and Rab5 endocytic proteins and on the USP8 deubiquitinating enzyme.

La drosophile se défend contre les infections microbiennes par un ensemble de réponses immunitaires très efficaces comme la synthèse de peptides antimicrobiens. L’expression de ces peptides antimicrobiens est contrôlée par deux voies indépendantes : la voie Toll et la voie IMD. La voie IMD est activée par PGRP-LC, une protéine de la famille PGRP (Peptidoglycan Recognition Protein). PGRP-LF est un régulateur négatif spécifique de la voie IMD. Il a été proposé que cette protéinepourrait agir spécifiquement au niveau du récepteur PGRP-LC, en séquestrant le ligand peptidoglycane (PGN) et en empêchant son accès à PGRP-LC. Mon travail de thèse a été de résoudre la structure des deux domaines PGRP de PGRP-LF, LFz et LFw, dans le but de caractériser le mécanisme de régulation par cette protéine.J’ai exprimé le domaine LFz dans des cellules S2 et le domaine LFw dans des bactéries. J’ai résolu la structure cristallographique de LFz à la résolution de 1.72Å et celle de LFw à la résolution de 1.94Å. Les structures de LFz et LFw montrent qu’elles ne possèdent pas la crevasse de liaison classique des PGRP, et ne peuvent pas interagir avec le PGN. J’ai confirmé ces résultats structuraux par des étudesbiochimiques de liaison des ces domaines à du PGN insoluble. L’aspect de régulation par PGRP-LF, par une séquestration du PGN, n’est donc valide. J’ai cloné et exprimé dans des cellules S2 les protéines PGRP-LCx et PGRP-LCa (partenaire du complexe activateur de la voie IMD) dans le but d’étudier leur interaction avec PGRP-LF. J’ai mis en évidence, par des analyses de résonnance plasmonique de surface, une interaction entre PGRP-LF et PGRP-LCx en absence et en présence duPGN. Ces données nous permettent de proposer un modèle dans lequel PGRP-LF assure la régulation négative de la voie IMD par compétition avec PGRP-LCa pour la liaison au récepteur PGRP-LCx. / The fruit fly defends itself against microbial infections by a set of highly effective immune responses that involve the synthesis of antimicrobial peptides. The expression of these antimicrobial peptides is controlled by two independent pathways: the Toll pathway and the IMD pathway. The IMD pathwayis activated by PGRP-LC, a protein of the PGRP family (Peptidoglycan Recognition Protein). PGRPLF is a specific negative regulator of the IMD pathway. It has been proposed that this protein specifically acts at the receptor PGRP-LC, by sequestering peptidoglycan (PGN) and preventing its access to PGRP-LC. My thesis work aims to solve the structure of the two PGRP domains of PGRPLF, LFz and LFw, in order to characterize the mechanism of regulation by this protein. I have expressed the LFz domain in S2 cells and the LFw domain in bacteria. I have solved the crystalstructure of LFz at 1.72 Å resolution and that of LFw at 1.94Å resolution. Structures of LFz and LFw show they do not possess the classical binding cleft found in others PGRP, and cannot interact with the PGN. I have confirmed these structural results with biochemical studies of binding of these domains with insoluble PGN. The model of regulation by PGRP-LF, by a sequestration of PGN, is no longer valid. I have cloned and expressed in S2 cells PGRP-LCx and PGRP-LCa proteins (partners of theactivating complex of the IMD pathway) in order to elucidate whether there is direct interaction between PGRP LF and one of two isoforms of PGRP-LC. I have demonstrated, through Surface Plasmon Resonance analysis, an interaction between PGRP-LF and PGRP-LCx. Actually, PGRP-LF regulates negatively the IMD pathway by competing with PGRP-LCa to bind to the PGRP-LCx receptor.

Energikriser och hållbarhet sätter en prägel på fastighetsbranschen och tvingar företagen att enligt lag göra åtgärder för att dra ner på energiförbrukningen. Redan 2012 kom energieffektiviseringsdirektivet (EED) och efter ändringar i direktivet 2019 fick principen om “energieffektivitet först” rättslig grund. Kort därefter blev det ett lagkrav för svenska företag att implementera IMD av tappvarmvatten i sitt fastighetsbestånd, men är IMD av tappvarmvatten verkligen ekonomiskt lönsamt och rätt väg ifall energieffektivitet är det slutliga målet? Studien syftar till att undersöka vilka är påföljderna av lagkravet från 1 juni 2022 gällande implementeringen av individuell mätning och debitering av tappvarmvatten i nyproduktion av flerbostadshus. Hur ser det faktiskt ut i praktiken och vilken avvägning fastighetsbolagen behöver göra gällande energieffektiva lösningar som bidrar till en förbättrad miljö kontra ekonomisk lönsamhet?   Vi har använt oss av en kvalitativ metod där det empiriska materialet grundar sig i sju semistrukturerade intervjuer med respondenter från några av de största fastighetsbolagen i Sverige som arbetar med flerbostadshus. Med hjälp av tidigare forskning och vår teoretiska referensram har vi kunnat analysera det empiriska materialet.  Resultatet visar att IMD av tappvarmvatten främst används som ett verktyg för att skapa rättvisa vid kostnadsfördelningen av vattenförbrukningen. De positiva påföljderna har lett till en ökad miljömedvetenhet bland hyresgästerna och sänkt vattenförbrukning, medan de negativa påföljderna är eventuella försämrade finansiella värden och ökade kostnader. / Energy crises and sustainability are leaving their mark on the real estate industry, forcing companies to take action to reduce energy consumption as required by law. The Energy Efficiency Directive (EED) was already introduced in 2012, and after amendments to the directive in 2019, the "energy efficiency first" principle gained a legal basis. Shortly thereafter, it became a legal requirement for Swedish companies to implement Individual Metering and Billing (IMD) in their property portfolios. However, is IMD truly economically viable and the right path if energy efficiency is the ultimate goal? This study aims to examine the consequences of the legal requirement from June 1, 2022, regarding the implementation of individual metering and billing for domestic hot water innew multi-dwelling buildings. What does the actual situation look like in practice, and what trade-offs do real estate companies need to make regarding energy-efficient solutions that contribute to an improved environment versus economic profitability? We have applied a qualitative method, where the empirical material is based on seven semi-structured interviews with respondents from some of the largest real estate companies in Sweden that work with multi-dwelling buildings. By using previous research and our theoretical framework, we have been able to analyze the empirical material. The results indicate that IMD for water is primarily used as a tool to promote fairness in the allocation of water consumption costs. The positive consequences have led to increased environmental awareness among tenants and reduced water consumption, while the negative consequences are potential negative impacts on financial values and increased costs.

Arboviruses are transmitted to vertebrates by arthropod vectors such as mosquitoes or ticks. The replication of Semliki Forest virus (SFV) (Togaviridae; Alphavirus) in vertebrate cells is well established and triggers cell death. SFV infection of Aedes albopictus mosquito cells was characterised. Virus growth curves were compared in three cell lines. Infection of U4.4 cells was persistent and did not affect growth of the culture. In contrast, infection of C6/36 and C7-10 cells resulted in a static culture with no cell division and no cell death. The response of U4.4 cells was characterised in greater detail using viruses containing fluorescent or luciferase markers within the replicase or structural open reading frame of the virus genome. Activation of the STAT/IMD pathway prior to SFV infection significantly reduced virus driven luciferase expression and virus production. Activation of the Toll pathway prior to SFV infection had no effect. However, activation of Toll in addition to STAT/IMD had a cumulative effect on luciferase expression and virus production. viRNAs were characterised by Illumina Solexa sequencing. Two percent of the small RNA species found in virus infected cells were derived from virus RNA. These were predominantly 21 nt long and mapped along the entire SFV genome and genome complementary RNAs. Generation of these viRNAs was not random. Some areas produced high frequencies and others no or very few; hot and cold spots respectively. There were no correlations between viRNA frequency and base pairing or secondary structures predictions. Cold spot-derived viRNAs were more effective than hot-spot viRNAs in inhibiting virus replication. Similar results were observed in Aedes aegypti-derived cells. Attempts were made to investigate the source of these viRNAs using a virus containing an IRES element which had been reported to prevent virus replication in insect cells but which did not efficiently do so in this study. A virus containing the RNAi inhibitor p19 was characterised and shown to increase virus production. Techniques for infecting mosquitoes via a blood meal feed were established. No infection was observed with virus replicon particles carrying a fluorescent marker gene. Infection was established using virus containing p19.

The existing UHF Satellite Communications (SATCOM) transponders used for military applications use efficient, saturated power amplifiers, which provide one earth-coverage antenna beam. The amplifier is dedicated to small frequency band and only handles a few carriers simultaneously. The communications capacity needed to support future military forces on the move will require satellite payload power amplifiers to support hundreds of channels simultaneously, with the channels spread over the entire military UHF SATCOM band. To meet the capacity requirements and simultaneously meet the out-of-band emission, power amplifiers will have to be highly linear. The high-efficiency, ultra-linear power amplifier architecture proposed to support the requirements can only be met by use of linearity improvement techniques. The literature search revealed many power amplifier linearity improvement techniques. Each technique was reviewed to determine its suitability for the proposed power amplifier architecture. The adaptive digital predistortion technique was found to be the most suitable in terms of bandwidth, correction achievable, and complication. A discussion on common linearization techniques is presented, followed by analysis of the adaptive digital predistortion technique. A SIMULINK simulation model of an adaptive digital predistorter was developed. The simulation results show that adaptive digital predistortion was able to significantly reduce the Inter-Modulation Distortion (IMD) terms generated by a memory-less power amplifier operating in the 240 MHz to 270 MHz range. An actual hardware implementation of adaptive digital predistorter was constructed and the test results show that there was a large reduction in IMD terms generated by a memory-less power amplifier. In the contrary, the results show there is only moderate improvement in IMD performance if the power amplifier has memory. The electrical memory in the power amplifier with memory was minimized, but this resulted only a modest improvement in the IMD performance. Therefore, it was concluded the majority of the memory effect was due to thermal memory.

All eukaryotic organisms have an innate immune system characterized by germ-line encoded receptors and effector molecules, which mediate detection and clearance of microbes such as bacteria, fungi, and parasites. VertebrateDrosophila as a genetically tractable organism with a This thesis concerns the peptidoglycan recognition protein (PGRP) gene family in the fruit fly. The family consists of thirteen genes, of which a few have been reported to be part of the signaling pathways that regulates immune Data presented show that the putative receptors have affinity for peptidoglycan, but not for lipopolysaccharide, or the fungal cell wall polymer beta-glucan. PGRP-SA, receptor of the Toll pathway, has a preference for In a search for novel PGRP receptors I found two PGRP proteins that instead displayed enzymatic activity towards peptidoglycan. They are of the N-actylmuramoyl L-alanine amidase type, which degrades peptidoglycan by splittingStaphylococcus aureus peptidoglycan looses its immune elicitor capacity. This is in contrast to lysozyme-degraded peptidoglycan, which isDrosophila PGRPs to be potential enzymes. PGRP-SB1 is the other enzymatic PGRP described within this thesis. It has a moreBacillus megaterium. In conclusion, receptor PGRP proteins binds bacterial peptidoglycan and triggers immune gene pathways and enzymatic PGRPs have the capacity to reduce the elicitor property of peptidoglycan.

In mobile communication new applications like wireless internet and mobile video have increased the demand of data-rates. Therefore, new more wideband systems are being implemented. Power amplifiers in the base-stations that simultaneously handle these wideband signals for many terminals (handhelds) need to be highly linear with a considerable band-width. In the past decade LDMOS has been the dominating technology for use in these RF-power amplifiers. In this work LDMOS transistors possible to fabricate in a normal CMOS process have been optimized and analyzed for RF-power applications. Their non-linear behavior has been explored using load-pull measurements. The mechanisms of the non-linear input capacitance have been analyzed using 2D TCAD simulations. The investigation shows that the input capacitance is a large contributor to phase distortion in the transistor. Computational load-pull TCAD methods have been developed for analysis of RF-power devices in high-efficiency operation. Methods have been developed for class-F with harmonic loading and for bias-modulation. Load-pull measurements with drain-bias modulation in a novel measurement setup have also been conducted. The investigation shows that the combination of computational load-pull of physical transistor structures and direct measurement evaluation with modified load-pull is a viable alternative for future design of RF-power devices. Simulations and measurements on the designed LDMOS shows a 10 to 15 % increase in drain efficiency in mid-power range both in simulations and measurements. The computational load-pull method has also been used to investigate the power capability of LDMOS transistors on SOI. This study indicates that either a low-resistivity or high-resistivity substrate should be used in manufacturing of RF-power LDMOS transistors on SOI to achieve optimum efficiency. Based on a proper substrate selection these devices exhibit a 10 % higher drain-efficiency mainly due to lower dissipated power in the devices.

This final year project describes the linearisation method Cartesian feedback and the design of such a feedback with a 2.4GHz power amplifier. To investigate the functionality of the Cartesian feedback ideal blocks with no current consumption were made and then gradually analog circuits were introduced into the feedback. The Cartesian feedback design consists of a subtracter, a modulator and a preamplifier in the top path and a demodulator and a filter in the feedback path. The blocks that are discussed in this report are the subtracter and the modulator unit. The circuits are designed in a 0.35µm SiGe BiCMOS technology. The result of the Cartesian feedback showed an increase in 1dB compression point by 6.2dBm and the IMD was improved by 17dB.

<p>This final year project describes the linearisation method Cartesian feedback and the design of such a feedback with a 2.4GHz power amplifier. </p><p>To investigate the functionality of the Cartesian feedback ideal blocks with no current consumption were made and then gradually analog circuits were introduced into the feedback. The Cartesian feedback design consists of a subtracter, a modulator and a preamplifier in the top path and a demodulator and a filter in the feedback path. The blocks that are discussed in this report are the subtracter and the modulator unit. The circuits are designed in a 0.35µm SiGe BiCMOS technology. </p><p>The result of the Cartesian feedback showed an increase in 1dB compression point by 6.2dBm and the IMD was improved by 17dB.</p>

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