Bacteria use quorum sensing, a cell to cell signaling mechanism mediated by small molecules that are produced by specific signal molecule synthases, to regulate gene expression in response to population density. In Vibrio anguillarum, the quorum-sensing phosphorelay channels information from three hybrid sensor kinases VanN, VanQ, CqsS that sense signal molecules produced by the synthases VanM, VanS and CqsA, onto the phosphotransferase VanU, to regulate activity of the response regulator VanO. VanO activates transcription of quorum-sensing regulatory RNAs (Qrr), which work together with the RNA chaperone Hfq to repress expression of the transcriptional regulator VanT. The work presented in this thesis characterizes quorum-sensing independent and quorum-sensing dependent mechanisms that regulate VanT expression. Moreover, an in vivo imaging system was established, as a means to study V. anguillarum infections in the rainbow trout infection model. Two quorum-sensing independent mechanisms regulating VanT expression were identified. First, the sigma factor RpoS indirectly activates VanT expression during transition into stationary growth phase by inhibiting hfq expression. Both, RpoS and VanT are crucial for stress response. Second, a type VI secretion system (T6SS) has a novel function as a signal sensing mechanism to regulate rpoS and vanT expression. Consequently, RpoS, quorum sensing and T6SS form a global network that senses stress and modulates stress response to ensure survival of the bacteria. Further analysis of the quorum-sensing dependent regulation of VanT expression by the phosphorelay system revealed that four qrr genes are expressed continuously during growth. The phosphotransferase VanU is suggested to activate two response regulators, VanO and a predicted second response regulator. Activated VanO induces expression of the Qrr sRNAs, whereas, the predicted response regulator represses expression of the Qrr sRNAs. Thus, VanU has a pivotal role in the regulation of VanT expression. The signal synthase VanM and VanT form a regulatory loop, in which VanM represses VanT by inducing expression of the Qrr sRNAs and VanT directly activates vanM expression to repress its own expression. Moreover, Hfq destabilizes vanM mRNA, repressing vanM expression. VanT forms another regulatory loop with the transcriptional regulator LuxT, in which LuxT activates vanT expression and VanT directly represses luxT expression. V. anguillarum is an opportunistic pathogen that causes vibriosis, a terminal hemorrhagic septicemia. The spatial and temporal progression of the infection was analyzed using the whole animal with an in vivo bioluminescent imaging method. Initial studies showed that colonization of the fish skin requires the siderophore, the RNA chaperone Hfq and the exopolysaccharide transport system, which protects against the innate immunity on the skin. Colonization of the fish skin is crucial for disease.

Denna uppsats syftar till att tydliggöra de mediala diskurser som finns om integration och utanförskap i tidningsartiklar. Tidningsartiklarna sträcker sig över två tidsperioder, 1996-1997 och 2006-2007. En frågeställning som uppsatsen försöker besvara är om analysens tidningsartiklar går att koppla till Ruth Levitas diskursmodeller om socialt utanförskap. Den centrala teoretiska utgångspunkten är ett postkolonialt perspektiv. Uttrycket ”vi och dem” utgör en röd tråd genom hela uppsatsen. Metoden för analysen är det diskursanalytiska verktyget diskursteori. Uppsatsens analys visar på tre centrala teman där samtliga utgår från utanförskap. Dessa teman är kultur och utanförskap, fördelning och utanförskap samt arbete och utanförskap.

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L'aroma és un factor de qualitat de la majoria dels aliments, ja que en depèn, en gran mesura, que el consumidor accepti o rebutgi el producte. En el vi i en les begudes alcohòliques esdevé, però, un dels factors més importants, ja que aquests productes no es consumeixen pel seu valor nutritiu, sinó pel plaer que en proporciona la degustació.L'anàlisi de l'aroma del vi no és un problema senzill de resoldre si es consideren els centenars de compostos volàtils que poden intervenir en la seva configuració aromàtica i els baixos nivells de concentració en què es troben molts d'ells. El treball d'investigació realitzat en aquesta tesi ha consistit en l'estudi i la caracterització de l'aroma del vi mitjançant l'aplicació de dues tècniques molt importants en aquest àmbit: la cromatografia de gasos amb detecció olfactimètrica (GCO) i el nas electrònic.La GCO és una tècnica imprescindible en l'estudi de l'aroma que, utilitzant el nas humà com a detector cromatogràfic, permet identificar, d'entre tots els compostos volàtils de la mostra, aquells que presenten olor i que, per tant, són susceptibles de participar en l'aroma. El treball més rellevant dut a terme amb aquesta tècnica ha estat el desenvolupament d'un mètode, per a caracteritzar l'aroma del vi, que utilitza la microextracció en fase sòlida (SPME) com a tècnica de pretractament de la mostra i que permet l'avaluació de la intensitat aromàtica dels diferents compostos mitjançant una nova modalitat de la tècnica de dilució AEDA (Aroma Extract Dilution Analysis).Els instruments coneguts com a nassos electrònics es van desenvolupar amb la finalitat d'imitar el nas humà en el control de qualitat que, des de temps immemorials, s'ha dut a terme mitjançant l'anàlisi sensorial. En l'anàlisi de begudes alcohòliques, però, els nassos electrònics han estat poc utilitzats degut als problemes de saturació que causa l'elevat contingut d'etanol als sensors de gasos que conformen la majoria d'aquests instruments. En aquesta tesi s'ha estudiat l'aplicació d'un nas electrònic basat en l'espectrometria de masses al control de qualitat del vi i de les begudes alcohòliques, concretament brandies i aiguardents de canya de sucre, ja que, a priori, l'etanol no suposa una limitació per a l'ús d'aquest instrument. La major part dels estudis realitzats s'han dirigit a l'avaluació del potencial d'aquesta tècnica en el control de la qualitat del vi. Amb aquest objectiu s'han dut a terme amb èxit diferenciacions de vins segons diversos paràmetres enològics, com són l'origen, la varietat o el temps d'envelliment. Per altra banda, s'ha fet un seguiment de la maduració del raïm i del procés d'elaboració del vi per tal d'avaluar la capacitat d'aquesta tècnica en el control d'aquests dos processos. Una altra aplicació estudiada ha estat la determinació de compostos amb un impacte sensorial important, com els defectes aromàtics no desitjats, mitjançant el calibratge multivariant. En aquesta línia s'han desenvolupat dos mètodes per a la determinació del 2,4,6-tricloroanisole, un compost que es troba en alguns vins a causa de contaminacions externes i que és el principal responsable del defecte aromàtic conegut com a "gust de suro". Per últim, s'ha estudiat també l'aplicació de l'instrument al control de qualitat dels aiguardents de canya de sucre amb el desenvolupament d'un mètode per a determinar el temps d'envelliment dels aiguardents en bótes de fusta de roure, així com també s'ha estudiat la correlació entre les dades obtingudes en l'anàlisi dels aiguardents amb el nas electrònic i les intensitats de diferents descriptors aromàtics avaluades per un panel de tastadors en una anàlisi sensorial de les mostres. / Aroma is an important factor in quality control and quality assurance of foods, but in wines and in alcoholic beverages, this factor is possibly the most important. Wine aroma analysis is not an easy task to perform because aroma is composed of complex mixtures of hundreds of volatile compounds with different sensory and chemical properties and, moreover, the concentration levels of most of them are very low.The research work carried out in this thesis lay in the study and the characterisation of wine aroma by two very important techniques in this field: gas chromatography olfactometry (GCO) and electronic nose. GCO is a very useful technique in the study of wine aroma that makes it possible to identify, among all the volatile compounds, the odorous volatile compounds of the sample by using the human nose as chromatographic detector. The most important study carried out in this thesis by GCO was the development of a method for the characterisation of wine aroma that uses the solid-phase microextraction technique (SPME) as a sample pre-treatment technique. This method evaluates the aromatic intensity of the odorous compounds by a new approach of the aroma extract dilution analysis (AEDA) technique.Quality control of food has been carried out by sensory analysis since immemorial time. However, sensory analysis by a panel of experts is a costly process for industries because it requires trained people who can work for only relatively short periods of time. The purpose of the development of electronic noses was to achieve an instrument that could mimic the human sense of smell and provide rapid sensory information (e.g., differences and similarities among samples, presence of aromatic defects, etc.). Food analysis is probably the field in which the greatest number of applications using electronic noses have been developed. However, in wine and alcoholic beverage research, few studies have been performed with this type of instrumentation because of the problems that ethanol causes in the gas sensors which compose most of the electronic noses. In this thesis, the application of a mass spectromety (MS) based electronic nose to wine analysis has been studied because, a priori, ethanol does not interfere in the analysis by this instrument. Some applications have also been carried out with alcoholic beverages as sugar cane spirits and brandies. The purpose of these studies was to evaluate the potential application of this technique in the quality control of this kind of samples. In wine analysis, this technique was successfully applied to differentiate and classify wines according to their origin, variety and ageing. It was also evaluated its capacity in the monitoring of the aroma profile evolution during grape ripening and alcoholic fermentation in order to study the potential use of this technique in the control of these processes. Other application studied was the determination of compounds with an important sensory impact, such as the off-flavours, by multivariate calibration. Two methods were developed to determine 2,4,6-trichloroanisole which is the main responsible of the aromatic defect known as "cork taint". In the quality control of sugar cane spirits, a method to determine the ageing time of these samples in oak wood barrels was developed. It was also studied the correlation between the electronic nose data and the intensity of several odour attributes evaluated by a panel of experts in a sensory analysis of the spirits.

“Shakespeare and the Drama of Politic Stratagems” focuses on how Shakespeare dramatically explores strategic issues similar to those discussed by Machiavelli and other early modern politic authors. The thesis is structured in order to tackle the diverse nature of strategy while developing and expanding on its most essential issues. The first chapter deals with the amoral and dangerous political world of the first tetralogy, a world in which one must be strategic in order to survive. Since not every strategist engages in the same kind of strategy or even agrees about what the best strategy might be, the second chapter outlines the different characteristics of Shakespeare’s strategists. These strategists can sometimes achieve success on their own, but no one can survive alone indefinitely, and the third chapter thus outlines the importance of strategic alliances and the dangers of making the wrong alliance. The fourth chapter deals with the numerous kinds of enemies that a strategist must contend with. Not all enemies fight in the same way, so a strategist must be on guard against an enemy’s deceptions, the focus of the fifth chapter. Even if these obstacles are overcome, even the most successful strategists will almost inevitably fail at some point or another. That failure may be due to some flaw in their schemes, or it may be due to the extreme difficulty of achieving success indefinitely. The final chapter deals with the perennial conflict between virtù and fortuna and thus the limits of politic stratagems. Machiavelli’s works can be seen as an epicenter of strategic thinking in the early modern period, and so they act as a guide through complex, contradictory, but ultimately rewarding issues of strategy and their consequences. Machiavelli serves as both analogue and foil, for while Shakespeare dramatizes similar strategic ideas, his dramatizations reveal greater truths about what is at stake when one explores the nature and consequences of politic stratagems. This thesis demonstrates the multiple factors that make strategy so dynamic and useful to a young dramatist in the process of discovering his own interests in the art of politics and the art of drama.

<p>The role of articular cartilage in diarthrodial joints is primarily mechanical as the tissue provides a nearly frictionless, load-bearing surface that supports and distributes forces generated during joint loading. Embedded within the extensive cartilage extracellular matrix (ECM), chondrocytes are surrounded by a narrow, distinct pericellular matrix (PCM) that is thought to regulate the biomechanical microenvironment of the cell and influence chondrocyte metabolism, cartilage homeostasis, and overall joint health. While previous studies of PCM mechanical properties required physical extraction of the cell and PCM from the tissue, atomic force microscopy (AFM) provides a means for high resolution microindentation testing that can be used to measure local mechanical properties in situ. This dissertation develops and applies AFM microindentation techniques to 1) evaluate the microscale elastic properties of the cartilage PCM and ECM in situ and 2) correlate site-specific biochemical composition with biomechanical properties of the PCM and ECM. </p><p>An AFM-based stiffness mapping technique was experimentally validated and applied to cartilage sections to evaluate ECM and PCM properties in situ with minimal disruption of native matrix integration. As expected, PCM elastic moduli were significantly less than ECM moduli, uniform with depth, and mechanically isotropic. ECM moduli exhibited distinct depth-dependent anisotropy and unexpectedly, were found to decrease with depth from the articular surface. Both the PCM and ECM exhibited alterations in microscale moduli and their spatial distributions when evaluated in cartilage presenting early degenerative changes associated with osteoarthritis (OA) as compared to healthy tissue. </p><p>The ability to correlate site-specific biochemical composition with local biomechanical properties provides a more complete characterization of the chondrocyte microenvironment. To this end, we developed novel immunofluorescence (IF)-guided AFM stiffness mapping and demonstrated that PCM mechanical properties correlate with the presence of type VI collagen. Extending this technique by using dual IF, we presented new evidence for a defining role of perlecan in the PCM, showing that interior regions of the PCM rich in perlecan and type VI collagen exhibit lower elastic moduli than peripheral PCM and ECM regions lacking perlecan. Furthermore, lower moduli at the PCM interior were significantly influenced by the presence of heparan sulfate. IF-guided AFM stiffness mapping was combined with enzymatic digestion to demonstrate that the micromechanical properties of the PCM exhibit high resistance to specific enzymatic digestion of aggrecan and aggrecan-associated glycosaminoglycans but are vulnerable to proteolytic degradation by leukocyte elastase. </p><p>Overall, this research generates new insights into the complex structural, compositional, and functional relationships between the cartilage ECM and PCM and provides the tools and framework for further studies to continue to investigate their importance in regulating chondrocyte physiology in health and disease.</p> / Dissertation

Intracellular bacteria have developed various mechanisms to enter and persist in host cells and, at the same time, to evade the host immune response. One such pathogen is Francisella tularensis, the etiological agent of tularemia. After phagocytosis, this Gram-negative bacterium quickly escapes from the phagocytic compartment and replicates in the host cell cytosol. For this mode of infection, several components of the Francisella pathogenicity island (FPI) are critical. Interestingly, some FPI proteins share homology to components of Type VI Secretion Systems (T6SSs), but their assembly and functionality remains to be shown in Francisella.The thesis focused on the characterization of several of these FPI components; more specifically, how they contribute to the infection cycle as well as their possible role in the putative T6SS. We identified three unique mutants, ΔiglG, ΔiglI and ΔpdpE, which to various degrees were able to escape the phagosomal compartment, replicate in the host cytosol and cause host cell cytotoxicity. In contrast, ΔiglE as well as mutants within the conserved core components of T6SSs, VgrG and DotU, were defective for all of these processes. In the case of IglE, which is a lipoprotein and localized to the outer membrane of the bacterial cell wall, residues within its N-terminus were identified to be important for IglE function. Consistent with a suggested role as a trimeric membrane puncturing device, VgrG was found to form multimers. DotU stabilized the inner membrane protein IcmF, in agreement with its function as a core T6SS component. The functionality of the secretion system was shown by the translocation of several FPI proteins into the cytosol of infected macrophages, among them IglE, IglC and VgrG, of which IglE was the most prominently secreted protein. At the same time, the secretion was dependent on the core components VgrG, DotU but also on IglG. Although we and others have shown the importance of FPI proteins for the escape of F. tularensis, it has been difficult to assess their role in the subsequent replication, since mutants that fail to escape never reach the growth-permissive cytosol. For this reason, selected FPI mutants were microinjected into the cytosol of different cell types and their growth compared to their replication upon normal uptake. Our data suggest that not only the metabolic adaptation to the cytosolic compartment is important for the replication of intracytosolic bacteria, but also the mechanism of their uptake as well as the permissiveness of the cytosolic compartment per se.

This thesis examines the role of interpretation in legal encounter in Lexington, Kentucky. Through an analysis of legal and interpretation practices, this study seeks to ascertain how these practices may affect non-native or low-proficiency English speakers’ (LLPs) experiences with both federal and local laws and legal spaces. This place-based study involves in-depth qualitative research. Using the methodological framework of feminist geo-jurisprudence, this research contributes to our understanding of 1) the limits of the publicity of legal space and, more specifically, the ways in which language barriers can prevent legal inclusion; 2) local strategies and tactics for dealing with the challenges to meaningful access before the law in terms of language as outlined by Title IV of the 1964 U.S. Civil Rights Act; 3) the broader implications of language access for immigrants and non-citizens at the intersection of legal discourse and society (discursive legal space). Furthermore, this research addresses the absence and presence of hospitality (Derrida, 2005) from this site of citizenship negotiation, and it addresses the ethics of hospitality behind the work that attempts to resist legal closure and to enforce laws that protect, rather than persecute, those facing language barriers.

A majority of low-income individuals living in public housing today are working or receiving some kind of assistance, but still struggle to make ends meet. Previous studies show that cost and availability are barriers to healthy eating for low-income individuals. The purpose of this study was to determine relationships among nutrition habits, health status, sources of income, and food and living resources for low-income residents in public housing. The study utilizes data collected over five years on the impact of the revitalization of the families. The sample was randomly selected from residents of the housing property in a Kentucky city. Results showed that low income is connected to limited access to healthy food options and individuals are more likely to be at risk for chronic health conditions such as diabetes or hypertension. When income and employment were low, families reported a greater rate of skipped meals, less consumption of daily meals, and more purchasing of high fat and sodium meals from convenience stores.

I denna studie har vi med hjälp av en hermeneutisk ansats studerat upplevelsen av etnisk diskriminering samt de strategier individer med dessa upplevelser använder för att hantera denna diskriminering. Vi har utfört kvalitativa intervjuer där vi intervjuat tio personer vilka själva beskriver sig ha upplevt etnisk diskriminering. Vårt syfte är att öka förståelsen för hur föreställningar om "den andre" påverkar den individ som inkluderas i denna kategori. Vi har tagit oss an vårt material med hjälp av teorier om kategorisering och exkludering och funnit att människor med en annan etnisk bakgrund än svensk upplever att etnisk diskriminering verkar exkluderande på arbetsmarknaden, något som påverkar deras liv som helhet genom en upplevelse av att ständigt behöva stå till svars för sina rättigheter och lika villkor. I vår huvudtolkning betraktar vi denna upplevelse som en rättegång där individer ständigt står åtalade. Brotten de åtalade står till svars för är på basis av "annanhet", vad som kännetecknar denna "annanhet" förändras ständigt och gör det omöjligt för individen att helt frikännas från åtalspunkterna.