Dissertationen zum Thema „Reception capacity“

Bats are a paragon of evolutionary success. They rely on parsimonious sensory inputs provided by echolocation, yet are able to master lives in complex environments. The outer ears (pinnae) of bats are intricately shaped receiver baffles that encode sensory information through a diffraction process. In some bat species with particularly sophisticated biosonar systems, such as horseshoe bats (Rhinolophidae), the pinnae are characterized by static as well as dynamic geometrical features. Furthermore, bats from these species can deform their pinnae while the returning ultrasonic waves impinge on them. Hence, these dynamic pinna geometries could be a substrate for novel, dynamic sensory encoding paradigms. In this dissertation, two aspects of this dynamic sensing process were investigated: (i) Do local shape features impact the acoustic effects during dynamic deformation of the bat pinna? and (ii) do these shape deformations provide a substrate for the dynamic encoding of sensory information? For this, a family of simplified biomimetic prototypes has been designed based on obliquely truncated cones manufactured from sheets of isobutyl rubber. These prototypes were augmented with biomimetic local shape features as well as with a parsimonious deformation mechanism based on a single linear actuator. An automated setup for the acoustic characterization of the time-variant prototype shapes has been devised and used to characterize the acoustic responses of the prototypes as a function of direction. It was found that the effects of local shape features did interact with each other and with the deformation of the overall shape. The impact of the local features was larger for bent than for upright shape configurations. Although the tested devices were much simpler than actual bat pinnae, they were able to reproduce numerical beampattern predictions that have been obtained for deforming horseshoe bat pinnae in a qualitative fashion. The dynamically deformable biomimetic pinna shapes were estimated to increase the sensory encoding capacity of the device by unit[80]{%} information when compared to static baffles. To arrive at this estimate, spectral clustering was used to break up the direction- and deformation-depended device transfer function into a discrete signal alphabet. For this alphabet, we could estimate the joint signal entropy across a bending cycle as a measure for sensory coding capacity. The results presented in this thesis suggest that bat biosonar posses unique dynamic sensing abilities which have no equivalent in man-made technologies. Sensing paradigms derived from bat biosonar could hence inspire new deformable wave-diffracting structures for the advancement in sensor technology.
Ph. D.

Cognitive radios have been proposed as a new technology to counteract the spectrum scarcity issue and increase the spectral efficiency. In cognitive radios, the sparse assigned frequency bands are opened to secondary users, provided that interference induced on the primary licensees is negligible. Cognitive radios are established in two steps: the radios firstly sense the available frequency bands by detecting the presence of primary users and secondly communicate using the bands that have been identified as not in use by the primary users.

In this thesis we investigate how to improve the efficiency of cognitive radio networks when multiple cognitive radios cooperate to sense the spectrum or control their interferences. A major challenge in the design of cooperating devices lays in the need for exchange of information between these devices. Therefore, in this thesis we identify three specific types of control information exchange whose efficiency can be improved. Specifically, we first study how cognitive radios can efficiently exchange sensing information with a coordinator node when the reporting channels are noisy. Then, we propose distributed learning algorithms allowing to allocate the primary network sensing times and the secondary transmission powers within the secondary network. Both distributed allocation algorithms minimize the need for information exchange compared to centralized allocation algorithms.
Doctorat en Sciences de l'ingénieur
info:eu-repo/semantics/nonPublished

This study aims to investigate challenges within the provision of integration support to quota refugees and municipal integration staffs understanding of these challenges in Skåne. It was conducted through questionnaires and semi-structured interviews with municipal integration staffs in Skåne. The research findings indicate three key challenges in the reception of quota refugees namely lack adequate housing, lack of financial resources from the region and government, and lack of provision of psychosocial support. Further challenges are connected to the lack of translators available to municipalities in the quota refugees’ mother tongue, the lack of English or major refugee languages among quota refugees, and often poor mental health which slow down Swedish language learning process and affect integration negatively. As its contribution, this study provides a broader view on challenges with the provision of integration support by municipalities regarding reception capacity, housing, and integration programs to quota refugees simultaneously. Thereby, it points out the differences among municipalities in terms of resource allocation for integration, as well as the political will to integrate quota refugees which create unequal chances for integration.

The University of Manchester, PhD by Published Work, 2018 For several reasons, small and medium enterprises (SMEs) are an important group of firms. In most market economies SMEs contribute significantly to wealth and job creation, economic growth, and product and service innovation. At the same time, SMEs are said to produce environmental impacts that are significant and that need managing and regulating. Their importance, from an economic and environmental perspective, is reflected in the fact that SMEs have become an established subject for research, with a distinct area of analysis focusing on how they manage their environmental impacts. Despite considerable interest in this area, aspects of their behaviour are in need of further examination, for there are still misunderstandings and gaps in knowledge. An area where gaps exist is how SMEs respond to different forms of environmental regulation (e.g., command-based or market-based approaches) and different forms of regulatory pressure (e.g., such as those pressures from civil society that might induce compliance-related activities or market forces that might flow through, and affect, the value chain). Why the gaps? On the one hand, and generally speaking, a common claim among those who have considered issues affecting smaller firms is that regulation is an important driver of environmental behaviour. There is a well-documented set of linked claims and empirical findings that smaller firms tend to be motivated by compliance with regulatory standards, yet owing to their scarce resources can find themselves hovering on the edge of compliance. Typically, SMEs will attempt to do no more than the law requires of them. They tend not, as it were, to go beyond compliance. Of course, this is an important observation - one that might say much, even if indirectly, about the motivations and intentions of smaller firms. It might indicate that SMEs, rather than addressing environmental issues, are more concerned with making cost savings and efficiency gains, or with satisfying the requirements of customers over such matters as product or service quality and delivery. While significant, there are at least three reasons why this view remains incomplete as an explanation for the interaction between SMEs, regulation and the environment. Firstly, this view tends to over-homogenise smaller firms. By treating them as a standardized group, the inference is that SMEs view and respond to regulation - i.e., they are all driven by regulation - in a broadly similar way. Secondly, it says little about how and why regulation drives behaviour. Claiming that regulation drives behaviour is helpful, but the claim is unduly narrow and leaves several important questions. In what ways does regulation drive behaviour? Does regulation drive all smaller firms in the same way? Thirdly, and finally, it suggests that different forms of regulation drive SME behaviour and that different forms of regulation drive this behaviour in broadly similar ways. That is, it is incomplete as it lacks appreciation of the widening scope of regulation and governance, and the nature of smart mixes of regulation. It fails to properly consider whether and how SMEs might respond differently to command-based regulation, market- or information-based measures, or self- or so-called civil regulatory pressures. On the other hand, and again in general terms, while those who have examined regulation have looked at how it can influence firms, they have tended to pay too little regard to how firms of different size may respond to different approaches or to how the factors and characteristics relating to size may shape the effectiveness of regulation. SMEs particularly are often discussed as an unusual sideshow that might raise different issues in relation, say, to the impacts of regulation on performance or innovation. That we often pay too little regard to how firms of different size may respond raises difficulties, particularly given our increasing understanding that there is no guarantee that a particular instrument will work in all situations. In other words, we are becoming more aware of the fact that the effectiveness or ineffectiveness of regulation is likely to be context-sensitive, and that the size of the enterprise is likely to be an important determinant of context. This thesis does take, and provides evidence for, the view that the organisational context is crucial to understanding how regulation functions. The thesis does not claim to provide all answers, but it does adopt the position that, in aggregate terms, a firm's size, or the factors that can be associated with size (e.g., resources, skills, knowledge, visibility, profile, stakeholder relations), and related factors concerning a firm's mind-set, can affect two things; first, the types of regulatory influences that may affect organizational behaviour and, second, how firms will respond to those influences. By focusing on SMEs, the thesis in some ways reinforces the dominant view that regulation is a driver of behaviour. Nevertheless, it goes much farther than this by showing, both theoretically and empirically, that there are important differences across SMEs and that these differences determine how and why they respond to regulation. It extends the common view by showing how SMEs differ not only in terms of the types of regulatory influences that shape their behaviour, but also in terms of how they react to these different influences. The emerging picture thus shows that the responses of firms are determined by their particular characteristics. The term used in this thesis is 'receptive capacity', which is shown to be a composite measure that includes the capabilities (e.g., resources, skills, knowledge) and orientations (e.g., views) of firms. It is suggested here that the range of receptive capacities across firms is enormous, since no two firms will be identical. Yet, it is argued - and demonstrated - that firms can be grouped according to certain identifiable characteristics, and that these groups of firms will respond to regulatory pressures in broadly similar ways; that is, there are groups of firms that have broadly similar resources and broadly similar worldviews. Thus, as well as suggesting that differences can be found at the micro level, it is demonstrated that there are sufficient commonalities across some firms that we can understand them as groups - groups of individual firms with some common characteristics. In conclusion, it is the differences across firms that provide us with a more sophisticated view of how SMEs are influenced by, and respond to, regulation. It is the nature of differences that is the main contribution of this research to both the fields of regulation and organisational and management studies. It is suggested finally that these differences have implications for how we design regulation, for how we may expect regulation to work or indeed not work, and for issues such as regulatory complexity and smart mixes.

ERRα est un récepteur nucléaire dont l’activité est controlée par des co-régulateurs transcriptionnels. La forte expression de ERRα dans les cancers est corrélée à un mauvais pronostic. Les mécanismes par lesquels ERRα régule la migration des cellules cancéreuses sont mal compris, tout comme les co-régulateurs impliqués. Nous avons identifié deux enzymes modificatrices d’histone, LSD1 et SET7, agissant comme régulateurs positifs de ERRα.I. ERRα modifie les activités biochimiques de la déméthylase LSD1 vers la déméthylation (activatrice) de H3K9me2. L’activation des cibles de ERRa-LSD1 (identifiées par RNA-Seq) requiert le recrutement de ce complexe aux sites d’initiation de la transcription (TSSs), réalisé par le facteur de transcription NRF1 qui, lui, ne régule pas l’activité enzymatique de LSD1.II. Un autre groupe de cibles de ERRα (identifié par RNA-Seq) est sous le contrôle de l’histone méthyltransférase SET7 qui mono-méthyle H3K4. Le recrutement de SET7 aux TSSs est contrôlé par le facteur de transcription ETS1, qui promeut les interactions entre SET7 et ERRα, conduisant à l’activation de l’expression des gènes en aval.Des analyses par Gene Ontology ont montré que les cibles communes de ERRα/LSD1 et de ERRα/SET7 sont fortement enrichies en termes d’invasion cellulaire. De manière cohérente, la déplétion individuelle de chacun de ces facteurs (et également celle de NRF1 ou ETS1) réduit les capacités d’invasion, observée en tests in vitro (transwell) ou in vivo par xénogreffe sur embryons de poisson-zèbre.En résumé, nos résultats montrent deux réseaux de régulation impliquant des modifications d’histone induites par ERRα, conduisant à l’invasion cellulaire
ERRα is a nuclear receptor whose activity mainly depends on its interaction with transcription co-regulators. High levels of ERRα are found in various cancer types and correlate with poor prognosis. However, the mechanisms linking ERRα to cancer cell migration as well as the coregulators involved are unclear. In our study, we found two histone-modifying enzymes, LSD1 and SET7, acting as positive regulators of ERRα.I. ERRα impacts the biochemical activities of the LSD1 demethylase. Activation of ERRα -LSD1 targets (identified by RNA-Seq) requires the recruitment of this complex at Transcriptional Start Sites (TSSs), which is achieved by the NRF1 transcription factor. In our study, we have shown several points: NRF1, but not ERRα , is involved in positioning LSD1 to TSS, whereas ERRα , but not NRF1, regulates LSD1 enzymatic activities towards demethylating H3K9me2.II. A distinct group of ERRa target genes (identified by RNA-Seq) is under the control of the histone methyltransferase SET7 which mono-methylates H3K4. Appropriate recruitment of SET7 at TSSs is controlled by the ETS1 transcription factor, promoting the interaction between SET7-ERRa, leading to target gene expression.Gene Ontology analysis revealed that ERRa-LSD1 co-targets, as well as ERRa-SET7 co-targets, are enriched in terms of cell invasion. Consistently, depletion of each of these factors, as well as depletion of NRF1 or ETS1, leads to reduced cell invasion capacities as observed in transwell assays or in vivo, using xenotransplantation in the zebrafish embryo.Altogether, our results show two regulatory networks involving histone modifications induced by nuclear receptors, leading to increased cell invasion

The main expansion of the discovery of genetic variants associated with complex diseases has occurred during the last decade. This expansion has been accompanied, and in some sense motivated, by the desire to use this information to improve the predictive capacity of many diseases with an unidentified familial component, including coronary heart disease (CHD), with the aim of translating this genetic knowledge into clinical practice. This doctoral thesis is structured in two lines of investigation that address distinct aspects of this issue, first to evaluate the possible role of genetic variation in a candidate gene in modulating CHD risk, and second to evaluate whether genetic information can be used to improve risk assessment tools used in clinical practice. In the first research line (described in Part I), we investigate the contribution of genetic variation in one of the most widely-studied genes in cardiovascular genetics, ESR1, which encodes the Oestrogen receptor α protein. We provide a solid meta-analysis of evidence regarding the most widely-studied variant in this gene and we further explore the role of a broad range of common and uncommon variants in this gene in CHD risk. Using these approaches, we find no evidence of association between the genetic variants studied and CHD risk. However, although we can confidently accept that common genetic polymorphisms are not associated with cardiovascular disease, we cannot discard the possibility that other types of variation in this gene (for instance epigenetic variation) could modify susceptibility to cardiovascular disease, or that other elements of this pathway are associated with an increased risk of CHD. In this research I have provided a reliable answer to this long running unanswered question in cardiovascular genetics, allowing research to re-focus on other elements of this system or other pathways. In the second line, we explored the possible utility of genetic information obtained from genome-wide association studies (GWAS) in prediction of 10-year risk of CHD events by adding this information to cardiovascular risk functions. We have followed the recommendations proposed by the American Heart Association for evaluating the utility of novel biomarkers in clinical practice, and have demonstrated that although the magnitudes of the effects of these genetic variants on CHD risk are modest, there is a tendency towards improvement in the capacity of the risk functions to predict future CHD events. The translation of genetic information into clinical practice was one of the main motivations for the investment in genome-wide association studies, and my research represents one of the first efforts to explore this possibility.
L’expansió principal pel que fa al descobriment de variants genètiques associades amb malalties complexes s’ha dut a terme durant la última dècada. Aquesta expansió ha estat acompanyada, i d’alguna forma motivada, pel desig d’usar aquesta informació per millorar la capacitat de predicció d’aquelles malalties on hi és present un cert component familiar però en les que no es coneixien les variants que conferien un major risc de patir la malaltia, entre elles la cardiopatia isquèmica (CI). La present tesis doctoral està estructurada en dues línies d’investigació que avaluen el possible rol d’un gen candidat en la susceptibilitat de la CI i també avalua la millora en la capacitat de predicció d’un esdeveniment coronari de les eines usades habitualment en la pràctica clínica mitjançant la inclusió d’informació genètica. Més concretament, la primera línea d’investigació es centra en la contribució de la variació genètica en un dels gens més estudiats en relació amb CI: el gen que codifica pel receptor d’estrogens alfa (ESR1). En aquesta línea hem proveït un sòlid meta-anàlisis entre la variant més àmpliament estudiada d’aquest gen i risc coronari i també hem explorat el paper de la majoria de les variants comunes descrites en aquest gen i risc de CI. Mitjançant cap dels anàlisis hem trobat evidència d’associació entre les variants genètiques en aquest gen i el risc de CI. No obstant això, i encara que podem acceptar que les variants genètiques comunes d’aquest gen no estan associades amb esdeveniments coronaris, no podem descartar que altres tipus de variació en aquest gen (com per exemple variació epigenètica) pugui estar modificant la susceptibilitat a patir un esdeveniment coronari, ni tampoc que altres elements de la mateixa cadena de senyalització estiguin associats amb la malaltia. En la segona línea d’investigació, hem explorat el possible paper de les variants genètiques, obtingudes mitjançant estudis d’associació global del genoma (GWAS), en la millora de la capacitat de predicció a 10 anys dels esdeveniments coronaris, mitjançant la seva addició en les funcions de risc cardiovascular clàssiques. Hem seguit les recomanacions proposades per la American Heart Association per l’avaluació en la pràctica clínica de nous biomarcadors, i hem demostrat que, tot i que la magnitud de l’associació d’aquestes variants és modesta, hi ha una tendència cap a la millora de la capacitat de predicció de les funcions de risc.

An important obstacle to a functional cure for HIV/AIDS is the persistence of viral reservoirs found throughout the body in various cells and tissues. Reservoirs can be latently infected cells, or in the case of follicular dendritic cells (FDC), non-infected cells that trap infectious virus on their surface through immune complexes (HIV-IC). Although several strategies have been employed to target and eliminate viral reservoirs, they are short-lived and ineffective. In an attempt to provide a long-term approach, chimeric antigen receptor T (CAR-T) cells were designed to eliminate native HIV on FDCs. Although effective at eliminating HIV-infected cells, and halting spreading infection, their ability to eliminate the viral reservoir found on (FDCs) remains unclear. We used a novel second-generation CAR-T cell expressing domains 1 and 2 of CD4 followed by the mannose binding lectin (MBL) to allow recognition of native HIV envelope (Env) to determine the capacity to respond to the viral reservoir found on FDCs. We employed a novel fluorescent lysis assay, the Carboxyfluorescein succinimidyl ester (CFSE) release assay, as well as flow cytometric based assays to detect functional CAR-T activation through IFN-γ production and CD107a (i.e., LAMP1) membrane accumulation to test cytolytic capacity and functional activation of CD4-MBL CAR-T cells, respectively. We demonstrated their efficacy at eliminating HIV-infected cells or cells expressing gp160. However, these CAR-T cells were unable to lyse cells bearing surface bound HIV-IC. We found that failed lysis was not a unique feature of a resistant target, but a limitation in the CAR-T recognition elements. CAR-T cells were inactive in the presence of free HIV or in the presence of concentrated, immobilized virus. Further experiments determined that in addition to gp120 recognition by the CAR-T, the adhesion molecule ICAM-1 was necessary for efficient CAR-T cell killing of HIV-infected cells. CAR-T cell activity and killing were inhibited in the presence of ICAM-1 blocking antibody. These results suggest that other factors, such as adhesion molecules, play a vital role in CAR-T responses to HIV-infected cells. In addition, our findings highlighted the necessity to consider all models of HIV reservoirs, including FDCs, when evaluating therapeutic efficacy.

The Toll-like receptor (TLR) family plays a crucial role in innate immunity by mediating the recognition of, and response to a variety of microorganisms. Dysregulation of TLR-mediated inflammatory responses may, however, lead to a variety of acute and chronic inflammatory conditions. The description of a naturally occurring soluble form of TLR2 (sTLR2), and the observation that sTLR2-depleted serum renders leukocytes hypersensitive to TLR2-mediated stimulation, demanded a full assessment of sTLR2's regulatory capacity and an investigation of the underlying mechanism and therapeutic potential. The present study addressed these issues, and reports that cells overexpressing sTLR2, or stimulated in the presence of the sTLR2 protein, are TLR2 hyposensitive. Regulation was TLR2-specific, affected NF-kB activation, phagocytosis and superoxide production. Mice administered sTLR2 with Gram-positive bacteria-derived components showed lower levels of the neutrophil (PMN) chemoattractant, keratinocyte-derived chemokine lower PMN numbers and late apoptotic PMN. Mononuclear cell recruitment was not affected, and endogenous peritoneal sTLR2 levels increased. Notably, the anti-inflammatory effect of sTLR2 did not compromise the capacity of mice to clear a live infection. sTLR2 interfered with the mobilisation of TLR2 to lipid rafts, acted as a decoy receptor, and disrupted the receptor (TLR2)-co-receptor (CD 14) interaction by associating with CD 14. In order to identify the region(s) of sTLR2 involved in the interaction with CD 14, the leucine-rich repeats (LRRs) of TLR2 were mutated, and the ability of these mutants to affect CD 14- dependent signalling was evaluated. Peptides representing the LRR6 and LRR20 were found to specifically inhibit CD14-dependent TLR2 triggering, indicating that these LRRs are directly involved in the TLR2-CD14 interaction. This study defines sTLR2 as an efficient regulator of TLR2-mediated inflammatory responses. The capacity of sTLR2 and TLR2-derived peptides to exert regulatory effects by targeting CD 14 may inform the design of therapeutics against inflammatory conditions that will aim at disrupting the co-receptor's activity, thus blunting, but not abrogating, microbial recognition and host responses.

Abstract Billions of reared fish are released to the wild to compensate e.g. for the loss of natural populations. However, the efficiency of the releases is low. It has been proposed that one of the factors affecting the low survival rate of reared fish is their low swimming capacity. The molecular, metabolic and structural characters of muscle fiber define the swimming capacity of fish. Swimming capacity is related to the ecological competence of the fish, including the ability to complete long migrations and catch pray. One of the aims of the current study is to compare the properties of muscles of reared and wild salmon. The second aim of the study is to alter the muscular parameters of reared fish closer to those of wild fish by means of training. The muscular differences between wild, reared and trained fish are analyzed with immunological, histochemical and electron microscopic methods. The main focus is on the dihydropyridine and ryanodine receptors. These receptors are involved for example in the initiation, force and velocity of muscle contraction. According to the results, the level of receptors is higher in the muscles of wild as compared to reared fish. The aerobic ATP production capacity is also higher in the wild fish. However, with training both the level of receptors and oxidative capacity of reared fish increase. Moreover, the swimming capacity is enhanced in trained fish, and there is a connection between the level of receptors and swimming capacity of fish. Training also affects the migration pattern of fish which starts to resemble more that of wild fish. In conclusion, the results of the current study show that the performance of fish as a whole depends on functional parameters at cellular level. For the first time, it is shown that the level of receptors involved in muscular contraction is low in muscles of reared fish. However, the muscular properties are not definite. It is now shown that with training, both the muscular and migration parameters of reared fish approach those of wild fish. This will most probably increase the survival probability of trained, reared fish in the future
Tiivistelmä Kalojen kasvatus ja istutus takaisin luontoon on yksi tärkeimmistä keinoista säädellä ja palauttaa kalakantoja vesistöihin. Maailmanlaajuisesti puhutaan miljardien kalojen istutusmääristä vuosittain. On kuitenkin hyvin tunnettu tosiasia, että kasvatetut kalat eivät selviä luonnossa yhtä hyvin kuin villit lajikumppaninsa. On arvioitu, että vain alle 5 % istutetuista kaloista selviää lisääntymisikään asti hengissä. Eräs tekijä, joka voi vaikuttaa kalojen selviytymiseen, on kalojen lihaskunto. Kasvatettujen kalojen uintikyvyn on todettu olevan heikko villeihin lajikumppaneihin verrattuna. Luonnossa kaloilta kuitenkin vaaditaan suurta uintikykyä esim. saalistukseen, pedoilta pakenemiseen ja vaellukseen. Eräs tämän työn päätavoitteista on määrittää, miten kasvatettujen ja villien kalojen lihasten molekulaariset, aineenvaihdunnalliset ja rakenteelliset ominaisuudet poikkeavat toisistaan, jotta voidaan arvioida mitkä solutason tekijät vaikuttavat kalojen uintikykyyn ja sitä kautta selviytymiseen. Toisaalta kasvatettujen kalojen lihasten toiminnallisten tekijöiden tasoja pyritään nostamaan harjoittelun avulla lähemmäksi villien vastaavaa ja täten vaikuttamaan kasvatettujen kalojen uintikykyyn ja sitä kautta lopulta selviytymiseen. Työssä lihasten ominaisuuksia analysoidaan immunologisin, histokemiallisin ja elektronimikroskooppisin menetelmin. Tutkimuksissa keskitytään erityisesti dihydropyridiini- ja ryanodiinireseptorien suhteellisiin määriin. Nämä reseptorit osallistuvat lihasten supistumisen aikaansaatiin ja niiden määrä korreloi positiivisesti lihasten voiman ja supistumisnopeuden kanssa. Tulosten mukaan villien kalojen lihaksissa on huomattavasti enemmän reseptoreita verrattuna kasvatettujen kalojen lihaksiin. Myös aerobinen ATP:n tuottokapasiteetti on villeillä kaloilla huomattavasti tehokkaampaa. Harjoittelun jälkeen kasvatettujen kalojen lihasten reseptorimäärät ja aerobinen kapasiteetti kuitenkin kasvavat lähemmäksi villien vastaavaa. Lisäksi lihasten reseptorimäärä ja uintikapasiteetti näyttävät korreloivan keskenään. Harjoittelun seurauksena kasvatettujen kalojen vaellusnopeus, eräs kalojen selviytymiseen vaikuttavista tekijöistä, muistuttaa myös enemmän villien vastaavaa. Yhteenvetona voidaan sanoa, että kalojen koko suorituskyky riippuu lihasten solutason mekanismien tehokkuudesta. Tässä työssä todettiin ensimmäistä kertaa, että kasvatettujen kalojen lihaksissa niiden reseptoreiden määrät, jotka liittyvät itse lihassupistuksen tehokkuuteen, ovat huomattavasti alemmat kuin villeillä. Lihasten toiminnalliset ominaisuudet eivät kuitenkaan ole muuttumattomia vakioita. Tulosten perusteella harjoitettujen kalojen sekä lihas- että vaellusominaisuudet lähestyvät villien vastaavaa. Tämä harjoittelun jälkeinen muutos lisää todennäköisesti kasvatettujen kalojen selviytymismahdollisuuksia

La maladie d'Alzheimer (MA) est une maladie neurodégénérative touchant plusieurs millions de personnes. La MA a une origine multifactorielle. Diverses études suggèrent qu'une perturbation du métabolisme du cholestérol contribue au développement de la MA. Cependant, la littérature présente beaucoup de confusion. Il est donc crucial de mieux caractériser le métabolisme et l'implication du cholestérol dans la MA. Ce travail s'est intéressé au récepteur Lipolysis Stimulated Lipoprotein Receptor (LSR) qui est un récepteur hépatique aux lipoprotéines participant à la clairance des lipides en phase post prandiale. Les objectifs de cette thèse ont été de caractériser la présence du récepteur LSR dans le cerveau de souris, de déterminer son rôle dans le contrôle de l'homéostasie du cholestérol cérébral et dans la physiopathologie de la MA. Ainsi, nous avons caractérisé la présence de LSR dans des structures cérébrales importantes pour les capacités cognitives et le métabolisme énergétique. Grâce à un modèle de souris hétérozygote pour le récepteur LSR, nous avons mis en évidence que la délétion d'un allèle LSR entraine une altération du métabolisme du cholestérol cérébral au cours du vieillissement, qui est corrélée avec une augmentation de la sensibilité au stress amyloïde. Ces résultats suggèrent un rôle du récepteur LSR dans le contrôle de l'homéostasie du cholestérol cérébral et renforcent l'idée qu'une altération de cette dernière peut impacter la physiopathologie de la MA. Enfin, nous avons observé que la déficience d'un allèle LSR chez des souris placées sous un régime hyperlipidique pouvait impacter le métabolisme lipidique périphérique ainsi que l'anxiété de ces souris
Alzheimer's disease (AD) is a neurodegenerative disease affecting millions of people. The origin of AD is multifactorial. Studies suggest that disturbance of cholesterol metabolism contributes to AD development. However, data in the literature is conflicting. It is therefore crucial to better characterize the metabolism and involvement of cholesterol in AD. This work focused on the Lipolysis Stimulated Lipoprotein Receptor (LSR), a hepatic lipoprotein receptor involved in the clearance of lipoproteins during the postprandial phase. The objectives of this thesis were to characterize LSR receptor expression profile in the mouse brain, and to determine its role in both brain cholesterol homeostasis and in the pathophysiology of AD. We identified and characterized LSR expression in brain structures that are involved in cognitive abilities and the regulation of energy metabolism. Next, using a mouse model heterozygous for the LSR receptor, we were able to demonstrate that the deletion of one allele LSR causes impaired brain cholesterol metabolism in aging, which was correlated with increased susceptibility to amyloid stress. These results suggest a role of LSR receptor in brain cholesterol homeostasis and show that alterations of the brain cholesterol metabolism can impact AD pathophysiology. Finally, we observed that the deficiency of an LSR allele in mice on a high fat diets affected peripheral lipid metabolism and the anxiety in these mice

In order to produce predictable and robust systems forprotein purification and detection, well characterized, small,folded domains descending from bacterial receptors have beenused. These bacterial receptors, staphylococcal protein A (SPA)and streptococcal protein G (SPG), possess high affinity to IgGand / or HSA. They are composed of repetitive units in whicheach one binds the ligand independently. The domains foldindependently and are very stable. Since the domains also havewellknown three-dimensional structures and do not containcysteine residues, they are very suitable as frameworks forfurther protein engineering.

Streptococcal protein G (SPG) is a multidomain proteinpresent on the cell surface ofStreptococcus. X-ray crystallography has been used todetermine the binding site of the Ig-binding domain. In thisthesis the region responsible for the HSA affinity of ABD3 hasbeen determined by directed mutagenesis followed by functionaland structural analysis. The analysis shows that the HSAbindinginvolves residues mainly in the second α-helix.

Most protein-based affinity chromatography media are verysensitive towards alkaline treatment, which is the preferredmethod for regeneration and removal of contaminants from thepurification devices in industrial applications. Here, aprotein engineering strategy has been used to improve thetolerance to alkaline conditions of different domains fromprotein G, ABD3 and C2. Amino acids known to be susceptibletowards high pH were substituted for less alkali susceptibleresidues. The new, engineered variants of C2 and ABD shownhigher stability towards alkaline pH. Also, very important forthe potential use as affinity ligands, these mutated variantsretained the secondary structure and the affinity to HSA andIgG, respectively. Moreover, dimerization was performed toinvestigate whether a higher binding capacity could be obtainedby multivalency. For ABD, binding studies showed that divalentligands coupled using non-directed chemistry demonstrated anincreased molar binding capacity compared to monovalentligands. In contrast, equal molar binding capacities wereobserved for both types of ligands when using a directed ligandcoupling chemistry involving the introduction and recruitmentof a unique C-terminal cysteine residue.

The staphylococcal protein A-derived domain Z is also a wellknown and thoroughly characterized fusion partner widely usedin affinity chromatography systems. This domain is consideredto be relatively tolerant towards alkaline conditions.Nevertheless, it is desirable to further improve the stabilityin order to enable an SPA-based affinity medium to withstandeven longer exposure to the harsh conditions associated withcleaning in place (CIP) procedures. For this purpose adifferent protein engineering strategy was employed. Smallchanges in stability due to the mutations would be difficult toassess. Hence, in order to enable detection of improvementsregarding the alkaline resistance of the Z domain, a by-passmutagenesis strategy was utilized, where a mutated structurallydestabilized variant, Z(F30A) was used as a surrogateframework. All eight asparagines in the domain were exchangedone-by-one. The residues were all shown to have differentimpact on the alkaline tolerance of the domain. By exchangingasparagine 23 for a threonine we were able to remarkablyincrease the stability of the Z(F30A)-domain towards alkalineconditions. Also, when grafting the N23T mutation to the Zscaffold we were able to detect an increased tolerance towardsalkaline treatment compared to the native Z molecule. In allcases, the most sensitive asparagines were found to be locatedin the loops region.

In summary, the work presented in this thesis shows theusefulness of protein engineering strategies, both to explorethe importance of different amino acids regarding stability andfunctionality and to improve the characteristics of aprotein.

Keywords:binding, affinity, human serum albumin (HSA),albumin-binding domain (ABD), affinity chromatography,deamidation, protein A, stabilization, Z-domain, capacity,protein G, cleaning-in-place (CIP), protein engineering, C2receptor.

L’objectif était d’évaluer l’efficacité de thérapies développées pour traiter la dystrophie musculaire de Duchenne (DMD, due à des mutations du gène de la dystrophine) dans la restauration de déficits cognitifs associés à ce syndrome. Deux pistes thérapeutiques visant à compenser les altérations cérébrales liées à la perte de dystrophine ont été explorées chez les souris mdx, modèle de DMD. Une approche pharmacologique basée sur la surexpression de l’utrophine, homologue de la dystrophine, n’améliore pas les déficits comportementaux des souris mdx. Par contre, une intervention génique basée sur l’épissage de l’exon muté conduit à la restauration d’une dystrophine endogène et une récupération d’altérations cérébrales comme l’agrégation des récepteurs GABAA et la plasticité synaptique hippocampique. Ceci suggère un rôle de la dystrophine dans la plasticité du cerveau adulte et l’applicabilité de cette approche de thérapie génique au traitement des altérations cognitives de la DMD
Therapies have been developed to treat Duchenne muscular dystrophy (DMD, due to mutation in the dystrophin gene), but their ability to restore the cognitive deficits associated with this syndrome has not been yet studied. We explored two therapeutic approaches to compensate for the brain alterations resulting from the loss of dystrophin in the mdx mouse, a model of DMD. A pharmacological approach based on the overexpression of utrophin, a dystrophin homologue, does not alleviate the behavioural deficits in these mice. In contrast, a genetic intervention based on the splicing of the mutated exon leads to the restoration of endogenous dystrophin and a recovery of brain alterations such as the clustering of GABAA receptors and hippocampal synaptic plasticity in mdx mice. These results suggest a role for dystrophin in adult brain plasticity and indicate that this gene therapy approach is applicable to the treatment of cognitive impairments in DMD

The demand for broadband wireless communication is growing rapidly, requiring more spectrum resources. However, spectrum usage is inefficient today because different frequency bands are allocated for different communication standards and most of the bands are not highly occupied. Cognitive radio systems with dynamic spectrum access improve spectrum efficiency, but they require wideband tunable receiver hardware. In such a system, a preselect filter is required for the RF receiver front end, because an out-of-band (OB) interferer can block the front end or cause distortion, desensitizing the receiver. In a conventional solution, off-chip passive filters, such as surface-acoustic-wave (SAW) filters, are used to reject the OB interferer. However, such passive filters are hardly tunable, have large area, and are very expensive. On-chip, high-selectivity, linearly tunable RF filters are, therefore, a hot topic in RF front-end research. Switched-capacitor (SC) RF filters, such as N-path filters, feature good linearity and tunability, making them good candidates for tunable RF filters. However, N-path filters have some drawbacks: notably, a poor harmonic response and limited close-by blocker tolerance. This thesis presents the design and implementation of several interferer-tolerant receivers based on SC technology. We present an RF receiver with a harmonic-rejecting N-path filter to improve the harmonic response of the N-path bandpass filter. It features tunable narrowband filtering and high attenuation of the third- and fifth-order LO harmonics at the LNA output, which improves the blocker tolerance at LO harmonics. The 0.2-1 GHz RF receiver is implemented in a 65 nm CMOS process. The blocker 1 dB compression point (B1dB) is -2.4 dBm at a 20 MHz offset, and remains high at the third- and fifth-order LO harmonics. The LNA’s reverse isolation helps keep the LO emission below -90 dBm. A two-stage harmonic-rejection approach offers a > 51 dB harmonic-rejection ratio at the third- and fifth-order LO harmonics without calibration. To improve tolerance for close-by blockers, we further present an SC RF receiver achieving high-order, tunable, highly linear RF filtering. We implement RF input impedance matching, N-path filtering, high-order discrete-time infinite-impulse response (IIR) filtering and downconversion using only switches and capacitors in a 0.1-0.7 GHz prototype with tunable center frequency, programmable filter order, and very high tolerance for OB blockers. The 40 nm CMOS receiver consumes 38.5-76.5mA, achieves 40 dB gain, 24 dBm OB IIP3, 14.7 dBm B1dB for a 30MHz blocker offset, 6.8-9.7 dB noise figure, and > 66dB calibrated harmonic rejection ratio. The key drawback of our earlier SC receiver is the relatively high theoretical lower limit of the noise figure. To improve the noise performance, we developed a 0.1-0.6 GHz chopping SC RF receiver with an integrated blocker detector. We achieve RF impedance matching, high-order OB interferer filtering, and flicker-noise chopping with passive SC circuits only. The 34-80 mW 65 nm receiver achieves 35 dB gain, 4.6-9 dB NF, 31 dBm OB-IIP3, and 15 dBm B1dB. The 0.2 mW integrated blocker detector detects large OB blockers with only a 1 us response time. The filter order can be adapted to blocker power with the blocker detector.

Yes
Seasonal animals undergo changes in physiology and behavior between summer and winter conditions. These changes are in part driven by a switch in a series of hypothalamic genes under transcriptional control by hormones and, of recent interest, inflammatory factors. Crucial to the control of transcription are histone deacetylases (HDACs), generally acting to repress transcription by local histone modification. Seasonal changes in hypothalamic HDAC transcripts were investigated in photoperiod-sensitive F344 rats by altering the day-length (photoperiod). HDAC4, 6 and 9 were found to change in expression. The potential influence of HDACs on two hypothalamic signaling pathways that regulate transcription, inflammatory and nuclear receptor signaling, was investigated. For inflammatory signaling the focus was on NF-κB because of the novel finding made that its expression is seasonally regulated in the rat hypothalamus. For nuclear receptor signaling it was discovered that expression of retinoic acid receptor beta was regulated seasonally. HDAC modulation of NF-κB-induced pathways was examined in a hypothalamic neuronal cell line and primary hypothalamic tanycytes. HDAC4/5/6 inhibition altered the control of gene expression (Fos, Prkca, Prkcd and Ptp1b) by inducers of NF-κB that activate inflammation. These inhibitors also modified the action of nuclear receptor ligands thyroid hormone and retinoic acid. Thus seasonal changes in HDAC4 and 6 have the potential to epigenetically modify multiple gene regulatory pathways in the hypothalamus that could act to limit inflammatory pathways in the hypothalamus during long-day summer-like conditions.
Biotechnology and Biological Sciences Research Council (BBSRC)