Yes / A novel dual-polarized patch antenna for ultra-wideband (UWB) applications is presented. The antenna consists of a square patch and four capacitively coupled feeds to enhance the impedance bandwidth. Each feed is formed by a vertical isosceles trapezoidal patch and a horizontal isosceles triangular patch. The four feeds are connected to the microstrip lines that are printed on the bottom layer of the grounded FR4 substrate. Two tapered baluns are utilized to excite the antenna to achieve high isolation between the ports and reduce the cross-polarization levels. In order to increase the antenna gain and reduce the backward radiation, a compact surface mounted cavity is integrated with the antenna. The antenna prototype has achieved an impedance bandwidth of 112% at (|S11| ≤ -10 dB) whereas the coupling between the two ports is below -28 dB across the operating frequency range. The measured antenna gain varies from 3.91 to 10.2 dBi for port 1 and from 3.38 to 9.21 dBi for port 2, with a 3-dB gain bandwidth of 107%. / IEEE Antennas and Propagation Society

Introduction: Sepsis impaired vascular integrity results in multiple organ failure. Circulating lactate level is positively correlated with sepsis-induced mortality. We investigated whether lactate plays a role in causing endothelial barrier dysfunction in sepsis. Methods: Polymicrobial sepsis was induced in mice by cecal ligation and puncture (CLP). Lactic acid was injected i.p. (pH 6.8, 0.5 g/kg body weight) 6 h after CLP or sham surgery. To elucidate the role of heat shock protein A12B (HSPA12B), wild-type, HSPA12B-transgenic, and endothelial HSPA12B-deficient mice were subjected to CLP or sham surgery. To suppress lactate signaling, 3OBA (120 μM) was injected i.p. 3 h before surgery. Vascular permeability was evaluated with the Evans blue dye penetration assay. Results: We found that administration of lactate elevated CLP-induced vascular permeability. Vascular endothelial cadherin (VE-cadherin), claudin 5, and zonula occluden 1 (ZO-1) play a crucial role in the maintenance of endothelial cell junction and vascular integrity. Lactate administration significantly decreased VE-cadherin, claudin 5, and ZO-1 expression in the heart of septic mice. Our in vitro data showed that lactate (10 mM) treatment disrupted VE-cadherin, claudin 5, and ZO-1 in endothelial cells. Mechanistically, we observed that lactate promoted VE-cadherin endocytosis by reducing the expression of HSPA12B. Overexpression of HSPA12B prevented lactate-induced VE-cadherin disorganization. G protein-coupled receptor 81 (GPR81) is a specific receptor for lactate. Inhibition of GPR81 with its antagonist 3OBA attenuated vascular permeability and reversed HSPA12B expression in septic mice. Conclusions: The present study demonstrated a novel role of lactate in promoting vascular permeability by decreasing VE-cadherin junctions and tight junctions in endothelial cells. The deleterious effects of lactate in vascular hyperpermeability are mediated via HSPA12B- and GPR81-dependent signaling.

Wireless and mobile communications are already playing an important role in our lives, and this will can only grow more and more due to the predominant importance and use of modern smartphones, tablets and any kind of connected devices. With this is mind, the spectrum for wireless and mobile communications is becoming incredibly overcrowded, leading to increasing requirements for RF front-end filters. This progress has encouraged an impressive need for developing low-cost, high performance, mass-producible, small footprint, and highly integrated front-end solutions for microwave and millimeter-wave systems and applications including emerging 5G and future wireless platforms. In this context, high quality factor resonators are usually typical basic building blocks of many high performance passive and active circuits, and its design has become even more challenging in the last decade. As a result, Substrate Integrated Waveguide (SIW) technology has attracted scientific community and industry attention as a very good candidate for developing such desired high-Q planar microwave devices. Recently, SIW is demonstrating to be a successful approach for implementing microwave and mm-wave filters with high Q-factor, easy integration with other planar circuits, and for mass-production manufacturing processes in many technologies (i.e. Printed Circuit Board (PCB) and Low Temperature Co-fired Ceramics (LTCC) technologies among them). Its enormous similarity with waveguides is probably one of the main reasons why the development of SIW-based components and circuits is rapidly growing among the research community. Other potential features that, combined with the former advantages, could be of huge interest in a wide range of wireless and mobile applications are a lively set of research subjects, such as compactness, advanced filtering responses, and recently frequency-agility capabilities. These key features have been recently introduced in the design of microwave filters for the next-generation wireless systems. Taking into account the above-mentioned background, the work carried out during the course of this PhD Thesis has been directed towards a further study of SIW technology to propose, analyze and develop an innovative and original resonator topology. The proposed topology is based on the extension of the classical coaxial waveguide resonator to SIW technology, and must take advantage of the characteristics of SIW devices to allow the design of improved and innovative microwave resonator filters for advanced wireless systems. This PhD Thesis includes the latest improvements made on this topic, from the working principles of the basic coaxial SIW block, until different applications for the design of compact quasi-elliptic and reconfigurable microwave filters. The results are promising and demonstrate the validity of the proposed topology for the design of high-Q microwave filters, as well as its potential application to implement complex designs. The general knowledge gained from these cases of study can be considered a good base for further developing this technology, which can help to improve its EM performance, and also contribute to a more general use in the market. / Las comunicaciones inalámbricas y móviles juegan un papel importante en nuestras vidas, y esto sólo puede ir a más debido a su enorme importancia y al uso de los modernos teléfonos inteligentes (del inglés, smartphones), tabletas y toda clase de dispositivos inalámbricos. Con todo esto en mente, el espectro electromagnético para comunicaciones inalámbricas y móviles se está saturando cada día más, lo que conlleva un constante aumento de los requisitos para los filtros de radio-frecuencia usados en las cabeceras de dichos sistemas. Este progreso ha llevado a un creciente interés en desarrollar componentes de microondas de bajo coste, alto rendimiento, pequeño tamaño, que permitan implementar soluciones altamente integradas para sistemas de alta frecuencia (i.e. microondas y ondas milimétrica) y sus aplicaciones, incluyendo entre ellas la emergente conexión 5G y las futuras plataformas inalámbricas. En este contexto, los resonadores de elevado factor de calidad constituyen generalmente los bloques básicos para el diseño de muchos circuitos pasivos (entre ellos filtros) y activos de alto rendimiento. Su diseño se ha convertido por tanto en un reto aún mayor en la última década. Como resultado de ello, la tecnología de guía de ondas integradas en substrato (Substrate Integrated Waveguide, SIW) ha atraído la atención de la comunidad científica e industrial, al revelarse como una buena aproximación para el desarrollo de dispositivos planares de microondas con excelentes prestaciones eléctricas, y en particular para la implementación de filtros de microondas y onda milimétrica de bajas pérdidas y elevada integración con circuitos en tecnología planar. Además, su flexibilidad se caracteriza también por su adecuación a diferentes procesos de fabricación y producción en masa, en tecnologías tales como los circuitos impresos (Printed Circuit Board, PCB) o la tecnología de materiales cerámicos multi-capa co-sinterizados a baja temperatura (Low Temperature Co-fired Ceramics, LTCC) entre otras. Su enorme similitud con las ya largamente estudiadas guías de onda es, probablemente, una de las principales razones por las cuales el desarrollo de dicho circuitos está creciendo rápidamente entre la comunidad de investigadores. Cabe mencionar como, además de las anteriores ventajas, otras características de la tecnología SIW que podrían ser de gran interés en una amplia gama de aplicaciones inalámbricas y móviles son la miniaturización, la posibilidad de implementar respuestas avanzadas de filtrado y, recientemente, las capacidades de sintonía en frecuencia de los componentes de microondas. De este modo, el trabajo desarrollado a lo largo de esta Tesis Doctoral se ha orientado hacia el planteamiento, análisis y desarrollo de una topología de resonador innovadora y original. Dicha topología se basa en una extensión de las cavidades coaxiales en guía de onda metálica a una implementación integrada en substrato inspirada en la tecnología SIW. Esta Tesis Doctoral recapitula los últimos avances que se han producido sobre este tema, empezando desde la descripción de los principios fundamentales de funcionamiento de las estructuras, hasta la demostración de varias aplicaciones concretas útiles para el diseño de filtros de microondas muy compactos, con respuestas filtrantes avanzadas y reconfigurables. Los resultados que se van a mostrar a continuación son prometedores, y demuestran la validez de la topología propuesta. El conocimiento general obtenido de los diferentes prototipos fabricados y caracterizados experimentalmente puede considerarse una buena base para seguir desarrollando esta tecnología, lo que puede ayudar a mejorar su rendimiento electromagnético, así como a contribuir a un uso más extendido de estos dispositivos en el mercado. / Les comunicacions sense fils i mòbils juguen un paper important en les nostres vides, i això només pot anar a més a causa de la gran importància i l'ús dels moderns telèfons intel·ligents (de l'anglès, smartphones), tablets i tota classe de dispositius sense fil. Tenint en compte tot açò, l'espectre electromagnètic per a comunicacions sense fils i mòbils s'està saturant cada dia més, el que comporta un constant augment dels requisits per als filtres de radiofreqüència usats en les capçaleres d'aquests sistemes. Aquest progrés ha portat a un creixent interès en desenvolupar components de microones de baix cost, alt rendiment, volum reduït, que permeten implementar solucions altament integrades per a sistemes d'alta freqüència (ie. microones i ones mil·limètriques) i les seves aplicacions, incloent l'emergent connexió 5G i les futures plataformes sense fils. En aquest context, els ressonadors d'elevat factor de qualitat constitueixen generalment els blocs bàsics per al disseny de molts circuits passius (entre ells filtres) i actius d'alt rendiment. El seu disseny s'ha convertit per tant en un repte encara més gran en l'última dècada. Com a resultat d'això, la tecnologia de guia d'ones integrades en substrat (Substrate Integrated Waveguide, SIW) ha atret l'atenció de la comunitat científica i industrial, al revelar-se com una bona aproximació per al desenvolupament de dispositius planars de microones amb excel·lents prestacions elèctriques , i en particular per a la implementació de filtres de microones i ones mil·limètriques de baixes pèrdues i elevada integració amb circuits en tecnologia planar. A més, la seua flexibilitat es caracteritza també per la seua adequació a diferents processos de fabricació i producció en massa, en tecnologies com ara els circuits impresos (Printed Circuit Board, PCB) o la tecnologia de materials ceràmics multicapa co-sinteritzats a baixa temperatura (Low Temperature Co-Fired Ceramics, LTCC) entre d'altres. La seua enorme similitud amb les ja llargament estudiades guies d'ona és, probablement, una de les principals raons per les quals el desenvolupament d'aquests circuits està creixent ràpidament entre la comunitat d'investigadors. Cal destacar com, a més de les anteriors avantatges, altres característiques de la tecnologia SIW que podrien ser de gran interès en una àmplia gamma d'aplicacions sense fils i mòbils són la miniaturització, la possibilitat d'implementar respostes avançades de filtrat i, recentment, les capacitats de sintonia en freqüència dels components de microones. Aquestes característiques clau s'han introduït recentment en el disseny de filtres microones per als sistemes sense fils de pròxima generació, convertint-se en objecte prioritari d'estudi per part de la comunitat científica. D'aquesta manera, el treball desenvolupat al llarg d'aquesta tesi doctoral s'ha orientat cap al plantejament, anàlisi i desenvolupament d'una topologia de ressonador innovadora i original. Aquesta topologia es basa en una extensió de les cavitats coaxials en guia d'ona metàl·lica a una implementació integrada a substrat inspirada en la tecnologia SIW. Aquesta tesi doctoral recapitula els últims avanços que s'han produït sobre aquest tema, començant des de la descripció dels principis fonamentals de funcionament de les estructures, fins a la demostració de diverses aplicacions concretes útils per al disseny de filtres i microones molt compactes, amb respostes de filtrat avançades i reconfigurables. Els resultats que es mostraran a continuació són prometedors, i demostren la validesa de la topologia proposada. El coneixement general obtingut dels diferents prototips fabricats i caracteritzats experimentalment es pot considerar com una bona base per seguir desenvolupant aquesta tecnologia, el que pot ajudar a millorar el seu rendiment electromagnètic, així com a contribuir a un ús més estès d'aquests dispositius en el mer / Sirci, S. (2017). Substrate Integrated Coaxial Filters with Fixed and Tunable Responses [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/78838

L'immunothérapie, en particulier l'utilisation d'inhibiteurs de PD-1 et de PD-L1, a révolutionné le traitement du cancer. Cependant, son efficacité varie, certains patients présentant une réponse limitée ou une rechute, souvent liée à une forte expression de PD-L1 dans le microenvironnement tumoral. Des recherches récentes suggèrent que les effecteurs de la voie Hippo, YAP et TAZ, pourraient jouer un rôle crucial dans la régulation de l'expression de PD-L1 dans les cellules cancéreuses, y compris celles du sein et du poumon. La voie Hippo, qui contrôle la survie cellulaire, la prolifération et la taille des organes, agit par l'inhibition médiée par la kinase Lats1/2 de YAP/TAZ. Lorsqu'ils ne sont pas phosphorylés, YAP/TAZ se déplacent vers le noyau, affectant l'expression de gènes tels que PD-L1 et PD-1. De plus, les récepteurs couplés aux protéines G (RCPG), y compris le récepteur de type 1 à l'angiotensine II (AT1), ont été identifiés comme des modulateurs potentiels de la réponse immunitaire dans le cancer. Cette étude vise à développer et valider de nouveaux biosenseurs basés sur la BRET pour observer l'activité de la voie Hippo en suivant le mouvement de YAP et de TAZ du cytoplasme au noyau et pour évaluer la capacité de ces capteurs à surveiller différents signaux stimulants, y compris la stimulation AT1R, offrant ainsi des aperçus de nouvelles stratégies thérapeutiques en immuno-oncologie. L'immunothérapie est apparue comme une approche révolutionnaire pour pallier les limites des traitements anticancéreux traditionnels, introduisant des médicaments ciblant les points de contrôle immunitaires, tels que les inhibiteurs de PD-1 (protéine de mort cellulaire programmée 1) et de PD-L1 (ligand de mort cellulaire programmée 1). Cependant, certains patients présentent une réponse inadéquate ou une rechute après traitement, notamment ceux avec une expression accrue de PD-L1 dans le microenvironnement tumoral. En conséquence, la modulation de PD-1 et de PD-L1 émerge comme une branche potentielle des thérapies immuno-basées. Des investigations récentes suggèrent que les effecteurs de la voie Hippo, YAP et TAZ, pourraient influencer la réponse immunitaire en régulant transcriptionnellement PD-L1 dans diverses cellules cancéreuses, y compris celles du sein et du poumon. Mécanistiquement, la kinase Lats1/2 inhibe YAP/TAZ par phosphorylation directe, les séquestrant dans le cytoplasme, tandis que YAP/TAZ non phosphorylés se transloquent dans le noyau, médiatisant la sortie transcriptionnelle de gènes tels que PD-L1 et PD-1. La signalisation des RCPG a été proposée comme un modulateur 4 potentiel de la réponse immunitaire au cancer, avec le récepteur de type 1 à l'angiotensine II (AT1) suggéré comme un médicament combiné potentiel pour les traitements en immunooncologie. Par conséquent, ce travail vise à développer et valider de nouveaux biosenseurs basés sur la BRET pour évaluer la voie Hippo en surveillant la translocation de YAP et de TAZ du cytoplasme au noyau et pour évaluer la capacité de ces capteurs à surveiller différents signaux stimulants, y compris la stimulation AT1R. Cette étude visait à développer et à valider des biosenseurs pour surveiller l'activité de la voie Hippo en évaluant la translocation de YAP et de TAZ à l'aide du transfert d'énergie de résonance bioluminescente améliorée (ebBRET). Des défis techniques ont entravé la validation complète sous différents stimuli, mais la manipulation de la densité cellulaire a validé la fonctionnalité du capteur. L'identification du récepteur AT1 comme un modulateur de la relocalisation de YAP médiée par les RCPG souligne la nécessité d'explorer l'interaction entre la signalisation des RCPG et la voie Hippo. La sélection minutieuse des activateurs et la validation complète sont cruciales pour élucider le réseau régulateur de YAP/TAZ dans le développement du cancer. / Immunotherapy, particularly the use of PD-1 and PD-L1 inhibitors, has revolutionized cancer treatment. However, its effectiveness varies, with some patients showing limited response or relapse, often linked to high PD-L1 expression in the tumor microenvironment. Recent research suggests that the Hippo pathway effectors, YAP and TAZ, may play a crucial role in regulating PDL1 expression in cancer cells, including breast and lung cancers. The Hippo pathway, which controls cell survival, proliferation, and organ size, operates through the Lats1/2 kinase-mediated inhibition of YAP/TAZ. When not phosphorylated, YAP/TAZ move to the nucleus, affecting the expression of genes like PD-L1 and PD-1. Furthermore, G protein-coupled receptors (GPCRs), including the Angiotensin II type 1 (AT1) receptor, have been identified as potential modulators of the immune response in cancer. This study aims to develop and validate novel BRET-based biosensors to observe the Hippo pathway's activity by tracking the movement of YAP and TAZ from the cytoplasm to the nucleus and to assess various stimulatory signals, including AT1R stimulation, offering insights into new therapeutic strategies in immuno-oncology Immunotherapy has emerged as a revolutionary approach to address the limitations of traditional cancer treatments, introducing drugs that target immune checkpoints, such as PD-1 (Programmed Cell Death Protein 1) and PD-L1 (Programmed Cell Death Ligand 1) inhibitors. Yet, some patients exhibit inadequate response or experience relapse post-treatment, especially those with heightened PD-L1 expression in the tumor microenvironment. Consequently, modulation of PD-1 and PD-L1 has emerged as a potential branch of immuno-based therapies. Recent investigations propose that the Hippo pathway effectors, YAP and TAZ, may influence the immune response by transcriptionally regulating PD-L1 in various cancer cells, including breast and lung cancer. Mechanistically, Lats1/2 kinase inhibits YAP/TAZ through direct phosphorylation, leading to their sequestration in the cytoplasm, while unphosphorylated YAP/TAZ translocate to the nucleus, mediating the transcriptional output of genes such as PD-L1 and PD-1. G proteincoupled receptors (GPCRs) signaling has been proposed as a potential modulator of the immune response to cancer, with the Angiotensin II type 1 (AT1) receptor suggested as a potential combination drug for immune-oncology treatments. Therefore, this work aims to develop and validate new BRET-based biosensors to evaluate the Hippo pathway by monitoring the 6 translocation of YAP and Taz from the cytoplasm to the nucleus and to evaluate the ability of these sensors to monitor different stimulatory signals, including AT1R stimulation. This study aimed to develop and validate biosensors for monitoring Hippo pathway activity by assessing YAP and TAZ translocation using enhanced bioluminescence resonance energy transfer (ebBRET). Technical challenges hindered comprehensive validation under different stimuli, yet cell density manipulation validated sensor functionality. Identification of AT1 receptor as a modulator of GPCR-mediated YAP relocalization underscores the need to explore GPCR-Hippo pathway interplay. Meticulous activator selection and comprehensive validation are crucial for unraveling YAP/TAZ's regulatory network in cancer development.

The present work focuses on the test method for the investigation of combined journal-thrust bearings. In simple applications, radial or thrust bearings are separated physically and can be considered as stand-alone machine elements. These can each be dimensioned by established calculation models. In compact designs, combined bearings must be used due to space savings. In this case, the bearing parts are present as a unit in which the axial part is supplied with lubricant from the radial part. If the bearing parts are treated separately, the oil outlet temperature of the radial part can serve as the input variable for the axial part. In this case, both the coupled thermal conduction between the running surfaces through the material and the pressure interaction between the two bearing parts are neglected. The thermal coupling of the bearing parts leads to an overall higher thermal load. The pressure interaction affects the respective volume flow, which also has a significant influence on the respective bearing temperatures. The development and validation of a simulation program required accompanying extensive measurements on the real bearing. For this purpose, the development of a suitable test bench for applying a combined radial and axial load is to be presented in this paper. In addition, the measurement procedures will be discussed, such as the determination of pressure and temperature in the bearing surfaces and the transition areas between journal and thrust bearing parts. Finally, the temperature distribution between a pure radial load on a journal bearing and a combined load on the identical journal-thrust bearing will be compared using the example of an operating point.

Increased salinity in freshwater systems – also called the Freshwater Salinization Syndrome (FSS) – can have far-ranging implications for the natural and built environment, agriculture, and public health at large. Such risks are clearly on display in the Occoquan Reservoir – a drinking water source for roughly one million people in the northern Virginia/ National Capital Region. Sodium concentrations in the Occoquan Reservoir are approaching levels that can affect taste and health. The Reservoir is also noteworthy as a flagship example of indirect potable reuse, which further adds complexity to understanding the sources of rising levels of sodium and other types of salinity. To help understand the role residential discharges might play in salinization of the Occoquan Reservoir, a suite of laundry detergent products was identified based upon survey data collected in the northern Virginia region. The ionic compositions of these products were then characterized using ion chromatography and inductively coupled plasma-mass spectrometry to quantify select ionic and elemental analytes. Sodium, chloride, and sulfate were consistently found in appreciable amounts. To comparatively characterize the laundry detergents, principal component analysis was employed to identify clusters of similar products. The physical formulation of the products was identified as a marker for their content, with dry formulations (free-flowing and encapsulated powders) being more enriched in sodium and sulfate. This result was corroborated by comparing nonparametric bootstrap intervals for individual analytes. The study's findings suggest an opportunity wherein consumer choice can play a role in mediating residential salt inputs in receiving bodies such as the Occoquan Reservoir. / Master of Science / Many streams, rivers, and other freshwater systems have become increasingly salty in recent decades. A rise in salinity can be problematic, stressing aquatic life, corroding pipes, and even enhancing the release of more pollutants into the water. This phenomenon, called Freshwater Salinization Syndrome, can threaten such systems' ability to serve as sources of drinking water, as is the case for the Occoquan Reservoir in northern Virginia. Serving roughly one million people, the Reservoir is notable for being one of the first in the country to purposely incorporate highly treated wastewater upstream of a drinking water supply. Despite the Reservoir's prominence, the reasons behind its rising salt levels are not well understood. This study sought to understand the role that individual residences could play when household products travel down the drain and are ultimately discharged into the watershed. Laundry detergents are potentially high-salt products. A survey of northern Virginian's laundry habits was conducted to understand local tastes and preferences. Informed by the survey, a suite of laundry detergents was chemically characterized to measure salt and element concentrations. The detergents were found to have notable amounts of sodium, chloride, and sulfate in particular, with sodium being the most abundant analyte in every detergent. However, not all detergents were equally salty; statistical tools revealed that dry formulations (such as powdered and powder-filled pak detergents) contributed more sodium and sulfate, among other things. This study's findings suggest that laundry detergents could be contributing to Freshwater Salinization Syndrome in the Occoquan Reservoir, and that local consumers' choice of detergents could make a difference.

Yes / Inhibitory GABAB receptors (GABABRs) can down-regulate most excitatory synapses in the CNS by reducing postsynaptic excitability. Functional GABABRs are heterodimers of GABAB1 and GABAB2 subunits and here we show that the trafficking and surface expression of GABABRs is differentially regulated by synaptic or pathophysiological activation of NMDA receptors (NMDARs). Activation of synaptic NMDARs using a chemLTP protocol increases GABABR recycling and surface expression. In contrast, excitotoxic global activation of synaptic and extrasynaptic NMDARs by bath application of NMDA causes the loss of surface GABABRs. Intriguingly, exposing neurons to extreme metabolic stress using oxygen/glucose deprivation (OGD) increases GABAB1 but decreases GABAB2 surface expression. The increase in surface GABAB1 involves enhanced recycling and is blocked by the NMDAR antagonist AP5. The decrease in surface GABAB2 is also blocked by AP5 and by inhibiting degradation pathways. These results indicate that NMDAR activity is critical in GABABR trafficking and function and that the individual subunits can be separately controlled to regulate neuronal responsiveness and survival. / BBSRC, MRC and the European Research Council

Der G Protein-gekoppelte Rezeptor 30 (GPR30) wurde vornehmlich im Kontext von schnellen Östrogeneffekten auf zelluläre Signaltransduktionskaskaden untersucht und stellt möglicherweise einen neuen Östrogenrezeptor dar. Die physiologische Funktion von GPR30 in vivo konnte jedoch bisher nicht ermittelt werden. Daher wurde in dieser Arbeit ein Gpr30-defizientes Mausmodell charakterisiert, bei dem ein Teil der kodierenden Sequenz durch einen LacZ-Reporter ersetzt wurde (Gpr30-lacZ). Die Integration des Konstruktes in den Gpr30-Locus wurde mittels Southern blotting und Real-time PCR verifiziert. Gpr30-positive Zelltypen wurden durch Kolokalisation von LacZ mit zelltyp-spezifischen Markerproteinen identifiziert. Weitere Versuche dienten der Aufklärung des Phänotyps von Gpr30-lacZ Mäusen. Zur Identifizierung von Proteinen des GPR30-Signalkomplexes wurden Yeast-Two-Hybrid Analysen mit der N- bzw. C-terminalen Domäne des Rezeptors durchgeführt. Die wesentlichen LacZ-positiven Zellpopulationen waren (i) Endothelzellen in kleinen arteriellen Gefäßen, (ii) glatte Muskelzellen, Perizyten und neuronale Subpopulationen im Gehirn, (iii) Hauptzellen in der Magenschleimhaut, (iv) Zellpopulationen in der Adenohypophyse und dem Hypophysenzwischenlappen sowie (vi) chromaffine Zellen im Nebennierenmark. Während der Phenotypisierung des Mausmodells wurde eine Reduktion der CD62L+ T-Zellen von ca. 50% im peripheren Blut festgestellt. Mittels Yeast Two-Hybrid Analyse wurden Pals1-associated tight junction protein (PATJ) und FUN14 domain-containing 2 (FUNDC2) als mögliche Interaktionspartner identifiziert. Zusammenfassend wurde in dieser Arbeit eine zelluläre Basis für die Funktion von Gpr30 in vivo ermittelt. Der Phänotyp in Gpr30-lacZ Mäusen ist wahrscheinlich durch eine verringerte Produktion von naiven T-Zellen im Thymus bedingt. PATJ bindet die C-terminalen Aminosäuren von GPR30 mit einer PDZ-Domäne und könnte ein Gerüst-protein des GPR30-Signal¬komplexes darstellen. / The orphan G protein coupled receptor 30 (GPR30) was predominantly analyzed in the context of membrane-initiated estrogen signaling suggesting that GPR30 represents a novel estrogen receptor. However, the physiological function of GPR30 in vivo remained unknown. To unravel the physiological role of murine Gpr30 in vivo, a Gpr30-deficient mouse model was analyzed that harbors a LacZ reporter (Gpr30-lacZ) within the Gpr30 locus. The targeting of Gpr30 was verified by Southern blotting and real-time PCR. Gpr30-expressing cell types were identified by colocalization of LacZ along with cell type-specific markers. Further experiments aimed to decipher the phenotype of Gpr30-lacZ mice. To gain information about the signaling complex of human GPR30, yeast two-hybrid screenings were performed with the N- and C-terminal domains as bait. The main LacZ-positive cell populations were (i) endothelial cells in small arterial vessels of various tissues, (ii) smooth muscle cells, pericytes, and neuronal subpopulations in the brain, (iii) gastric chief cells in the stomach, (iv) cells in the intermediate and anterior pituitary, and (v) chromaffin cells in the adrenal glands. Extensive phenotype screening at the German Mouse Clinic revealed reduced numbers of T cells in the peripheral blood of Gpr30-lacZ mice. Especially the proportion of CD62L+ cells was decreased by approx. 50%. Yeast two-hybrid screening led to the identification of Pals1-associated tight junction protein (PATJ) and FUN14 domain-containing 2 (FUNDC2). In conclusion, this study provides a cellular basis for the function of Gpr30 in vivo. Since CD62L+ cells represent the naive T cell compartment, the phenotype of Gpr30-lacZ mice suggests an impaired production of T cells in the thymus. PATJ likely binds the C-terminus of GPR30 with one of its PDZ domains and may represent a scaffolding protein of the GPR30 signaling complex.

An enhanced thermal imaging concept made possible through the development of a gradient-indexed metamaterial infrared detector that offers broadband transmission and reflection in THz waves. This thesis proposes a proof of feasibility for a metamaterial infrared detector containing an anti-reflective coating with various geometrically varying periodic metasurfaces, a gradient-indexed dielectric multilayer for near-perfect longpass filtering, and a gradient index of refraction (GRIN) metalens for enhanced focal plane thermal imaging. 2D Rigorous Coupled-Wave Analysis (RCWA) is used for understanding the photonic gratings performance based on material selection and varying geometric structure. Finite Difference Time Domain (FDTD) is used to characterize performance for a diffractive metalens by optimizing the radius and arrangement of cylindrical nanorods to create a desired phase profile that can achieve a desired focal distance for projections on a detector for near- to far-infrared thermal imaging. Through combining a micromachined anti-reflective coating, a near-perfect longpass filter, and metamaterial GRIN metalens, infrared/THz focal plane thermal imaging can obtain faster photo-response and image quality at targeted wavelengths, which allows for scientific advancements in electro-optical devices for the Department of Defense, aerospace, and biochemical detection applications.

Humane Zytomegalieviren (HCMV) etablieren nach einer Primärinfektion eine lebenslange latente oder persistierende Infektion. Es wird allgemein angenommen, daß hieran die Manipulation der humanen Immunantwort durch das Virus beteiligt ist. Hierzu zählen die Hemmung von zytotoxischen CD8+ T-Zellen durch das Genprodukt US11 und die Beeinträchtigung der Leukozytenwanderung durch die Hemmung des Chemokinsystems durch den Chemokinrezeptor US28. Die Effizienz der US11-vermittelten Hemmung der T-Zell-Aktivierung wurde in einem rekombinanten Modell zur MHC-Klasse-I-vermittelten T-Zell-Aktivierung untersucht. Obwohl die Expression der MHC-Klasse-I-Moleküle durch US11 in dendritische Zellen (DCs) um bis zu 60% vermindert war, konnte keine Hemmung der T-Zell-Proliferation beobachtet werden. US28 ist der einzige funktionelle Rezeptor für die inflammatorischen Chemokine MCP-1, MCP-3, RANTES, MIP-1(, MIP-1( sowie Fraktalkine. Er kann sowohl Liganden-abhängig die Aktivierung von MAPK als auch die konstitutive Aktivierung von NF-(B vermitteln. In der vorliegenden Arbeit konnte mit Hilfe einer Rezeptormutante der Argininrest an Position 129 des DRY-Motivs als Voraussetzung für die Aktivierung der Signalwegen identifiziert werden. Ferner bewirkt die Expression des US28-Rezeptors die Entfernung inflammatorischer Chemokine aus der Umgebung infizierter Zellen. Molekulare Grundlage der Liganden-Depletion stellt die Endozytose des US28-Liganden-Komplexes dar. Es konnte gezeigt werden, daß der US28-Rezeptor eine Umlagerung von (-Arrestin-Molekülen in Vesikel vermittelt, jedoch unabhängig von Arrestin-Molekülen endozytiert wird. Die Endozytose des US28-Rezeptors war abhängig von der GTP-ase Dynamin. Ebenso konnte die Beteiligung des Lipid-Raft-Weges an der US28-Endozytose gezeigt werden. Die Hemmung des Clathrinweges bewirkte jedoch eine zweifach stärkere Verminderung der US28-Endozytose, kann der Clathrin-abhängige Weg als der Hauptweg der US28-Endozytose angesehen werden. / Primary infections of the human cytomegalovirus (HCMV) are followed by a lifelong infection in the state of latency or persistence. It is believed that the virus employs a number of immunomodulatory mechanisms to establish latent infections. Among these are the inhibition of cytotoxic CD8+ T-cells by US11 and the impairment of leukocyte migration by US28. The potency of US11 to mediate the inhibition of T-cell activation was analysed in a model of MHC class I mediated T-cell activation. Surface expression of MHC class I molecules was reduced by 60 % after expression of US11 in murine dendritic cells. In contrast, there was no reduction in the capacity of the dendritic cells to induce T-cell proliferation. The US28 gene product has been characterized as a functional receptor for the inflammatory chemokines RANTES, MCP-1, MCP-3, MIP-1?? MIP-1? and fractalkine.Upon ligand stimulation US28 mediates the activation of MAPK and additionally a constitutive activation of NF-?B. By generating site directed receptor mutant it was shown that the arginine at position 129 represents a structural requirement for both the ligand-induced and the constitutive signaling by US28. Moreover, it was suggested that the US28 dependent sequestration of chemokines from the environment of infected cells hinders leukocytes from the recruitment to sites of viral infection. A molecular mechanism for the ligand depletion is provided by the endocytosis of US28-ligand complexes. Studies revealed that US28 expression induced a redistribution of ?-arrestin molecules into vesicular structures but was dispensable for the endocytosis of the US28 receptor. However, US28 internalization was dependent on the small GTPase dynamin and by impaired receptor endocytosis after inhibition of the lipid raft pathway. Since inhibition of the clathrin dependent pathway resulted in a two-fold stronger reduction of US28 endocytosis, the clathrin-dependent pathway can be considered as the major route of US28 endocytosis.