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Role of methionine sulfoxide reductase in thermal-induced spreading depression coma in Drosophila melanogasterUnknown Date (has links)
Drosophila melanogaster encounter periods of increased temperature or decreased oxygen in its native environment. One consequence of these environmental stresses is increased production of reactive oxygen species that damage major molecules within cells. Another consequence is that flies fall into a protective coma where biological functions are minimized to conserve energy expenditures. This biological phenomenon is called spreading depression. The overarching aim of this project is to determine if methionine sulfoxide reductases affect entrance or exit from the protective coma induced by acute thermal stress. The data revealed that complete deficiency of Msr in young flies causes a faster induction of the coma. In both young and old flies, Msr does not affect average recovery time but does affect the pattern of recovery from coma. Entrance into the coma is age dependent with young flies maintaining activity longer than before entering into the coma as compared to old flies. / by Karin Schey. / Thesis (M.S.)--Florida Atlantic University, 2012. / Includes bibliography. / Mode of access: World Wide Web. / System requirements: Adobe Reader.
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Modelagem de medidas de controle em redes de movimentação de animais / Modeling control measures in networks of animal movementsOssada, Raul 28 August 2015 (has links)
A movimentação de animais em uma rede de fazendas e o espalhamento de algumas doenças animais estão intrinsecamente relacionados. Assim, compreender a dinâmica do espalhamento de doenças infecciosas nestas redes é um instrumento importante no controle dessas doenças. Usando as informações sobre as movimentações de bovinos no estado de Mato Grosso, Brasil, em 2007, reconstruiu-se a rede de trânsito e a rede de proximidade geográfica entre os estabelecimentos desse estado, além de redes hipotéticas seguindo os modelos de rede Molloy-Reed, Kalisky, Método A e Método B, onde simulou-se, usando diferentes configurações do modelo SLIRS, o espalhamento de doenças com parâmetros hipotéticos e reais (brucelose e febre aftosa). Além disso, simulou-se o controle do espalhamento dessas doenças considerando o controle por imunização e por restrição, com e sem rearranjo das movimentações após a restrição, selecionando os estabelecimentos a serem protegidos de forma aleatória, baseando-se no grau de movimentação dos animais e utilizando o conceito do paradoxo da amizade. Dentre os resultados, destacam-se que apesar dos padrões das curvas de prevalência nas redes hipotéticas serem semelhantes aos da rede real, os valores observados foram maiores nas redes hipotéticas, indicando que utilizá-las no planejamento de políticas de controle de doenças no lugar da rede real pode levar a um maior uso de recursos do que seria necessário. Além disso, no controle das doenças tanto com parâmetros hipotéticos quanto com parâmetros reais, nas simulações usando apenas a rede de trânsito dos animais, observou-se uma redução mais efetiva da prevalência ao se selecionar os estabelecimentos com maior grau total do que a da seleção aleatória, enquanto que nas simulações que consideraram a rede de proximidade geográfica dos estabelecimentos, a redução na prevalência das estratégias que selecionaram estabelecimentos específicos foram semelhantes aos da seleção aleatória. Sobre o efeito do rearranjo das movimentações, observou-se que este pode facilitar o espalhamento de doenças na rede, mesmo nas situações em que se aplica alguma estratégia de controle. Espera-se que os resultados das simulações matemáticas possam contribuir para a discussão do impacto relativo entre as estratégias de controle mencionadas e que futuramente possam auxiliar nas atividades dos órgãos responsáveis pela vigilância epidemiológica e no desenvolvimento de políticas de prevenção e controle de doenças em animais. / The animals’ movements in a farms network and the spread of some animal diseases are intrinsically related. Therefore, comprehending the dynamics of the spreading of infectious diseases in these networks is an important tool in controlling these diseases. Using the information about the bovine movements from the State of Mato Grosso, Brazil, in 2007, we rebuilt the network of animal movements and the geographic proximity network between the premises of this state, in addition to hypothetical networks following the network models Molloy-Reed, Kalisky, Method A and Method B, where we simulated, using different configurations of the model SLIRS, the spread of diseases with hypothetical parameters e real ones (brucellosis and foot and mouth disease). Moreover, we simulated the control of these diseases spreading, considering the control by immunization and by restriction, with and without the rearrangement of the movements after the restriction, selecting the premises to be protected randomly, based on the degree of animal’s movements and using the concept of the friendship paradox. Among the results, stands out that although the pattern of the prevalence curves in the hypothetical networks were similar to the ones in the real network, the observed values were higher in the hypothetical networks, indicating that using them in the planning of policies to control diseases in place of the real network might lead to a greater expense of resources than it would be necessary. Furthermore, in the control of the diseases both with hypothetical parameters as well as with real parameters, in the simulations using only the animal’s movements network, it was observed a more effective reduction of the prevalence when selecting the premises with the highest total degree than the random selection, while in the simulations that considered the network of geographic proximity of the premises, the reduction in the prevalence of the strategies that selected specific premises were similar to the random selection. On the effect of rearranging the movements, it was observed that it may facilitate the spread of diseases in the network even in situations where some control strategy is used. We hope that the results of the mathematical simulations may contribute to the discussion of the relative impact of the mentioned control strategies and that in the future they may assist in the activities of agencies responsible for disease surveillance and in the development of policies to prevent and control diseases in animals.
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Propriedades superficiais de filmes à base de gelatina / Surface properties of gelatin-based filmsPulla Huillca, Palmer Vicente 03 December 2015 (has links)
O objetivo do presente trabalho foi caracterizar as propriedades superficiais de filmes à base de gelatina. Para o qual foram elaborados filmes de: (i) Gelatina plastificado com glicerol (G) (gelatina: 5 g/100 g SFF; glicerol: 30 g/100 g de gelatina), (ii) Gelatina reforçado com montmorilhonita (G/MMT) (gelatina: 5 g/100 g SFF; glicerol: 30 g/100 g de gelatina; MMT: 5 g/100 g de gelatina) e Gelatina plastificado com citrato de acetiltributila (G/ATB) (gelatina: 5 g/100 g SFF; ATB: 50 g/100 de gelatina; lecitina de soja: 60 g/100 g de ATB; etanol: 20 g/100 g SFF). Os filmes foram produzidos mediante o uso de um aplicador automático de filmes \"Spreading\". Logo, os filmes foram submetidos a testes para determinação da espessura, umidade e propriedades óticas (brilho, cor e opacidade). Também foi caracterizada a microestrutura por microscopia eletrônica de varredura (MEV) e microscopia de força atômica (AFM); às imagens obtidas por MEV foi aplicado um analise de imagem mediante o programa Image J, para obter o valor da dimensão fractal (DF). Depois foram caracterizadas as propriedades superficiais de ângulo de contato (AC), molhabilidade ou coeficiente de espalhamento (Se), e energia livre superficial (ELS) mediante a medida do ângulo de contato pelo método da gota séssil (água: 5 µL e 1-Bromonaftaleno: 3 µL). Para o cálculo da ELS também foi aplicado o método de Owens-Wendt. Estas caracterizações foram feitas em ambos os lados do filme, lado ar e lado placa. A natureza do filme de G/ATB influenciou na umidade e as propriedades óticas, enquanto que os filmes de G e G/MMT apresentaram características similares. Em relação à microestrutura e rugosidade, o filme de G apresentou a superfície mais homogênea e lisa, contrario ao observado no filme de G/MMT, que apresentou a maior rugosidade seguida do filme de G/ATB. Foi observado que houve uma relação entre os valores de rugosidade e DF. De acordo com o valor do AC, os filmes apresentaram um caráter hidrofóbico, pois seus valores foram superiores a 65° (em ambos os lados dos filmes), na seguinte ordem: G/MMT > G > G/ATB; sendo que o Se seguiu a mesma tendência. Cabe mencionar também que não foi encontrada uma correlação significativa entre os valores de AC e rugosidade. Em função dos valores de AC, Se e ELS (especificamente a componente polar), o filme de G/ATB apresentou o menor caráter hidrofóbico, pois apresentou menores valores de AC e maiores valores de Se em comparação com os outros dois filmes. Os valores da componente polar da G/ATB foram os maiores, explicando de melhor maneira o caráter menos hidrofóbico deste filme. Pode-se concluir que os filmes à base de gelatina elaborados no presente trabalho têm propriedades hidrofóbicas (AC>65°), sendo a G/MMT o filme com melhores características hidrofóbicas. / The aim of this study was to characterize the surface properties of gelatin-based films. For which, films were produced as following: gelatin plasticized with glycerol (gelatin: 5 g/100 g SFF; glycerol: 30 g/100 g of gelatin), gelatin reinforced with montmorillonite (gelatin: 5 g/100 g SFF; glycerol: 30 g/100 of gelatin; montmorillonite: 5 g/100 g of gelatin), and gelatin plasticized with acetyltributyl citrate (gelatin: 5 g/100 g SFF; acetyltributyl citrate: 50 g/100 of gelatin, soy lecithin: 60 g/100 g acetyltributyl citrate; ethanol: 20 g/100 g SFF). The films were produced by using an automatic film applicator \"Spreading\". Then, the films were tested for determining thickness, humidity and optical property (gloss, color and opacity). Also the microstructure was characterized by scanning electron microscopy (SEM) and atomic force microscopy (AFM); in the images obtained by SEM it was applied image analysis using Image J program, to get the value of the fractal dimension (FD). After, the surface properties of contact angle (CA), wettability or spreading coefficient (Cs) and surface free energy (SFE) were characterized by measuring the contact angle by the method of the sessile drop (water: 5 µL, 1-Bromonaphthalene: 3 µL). For the calculation of the ELS, it was also applied the method of Owens-Wendt. These characterizations were made on both sides of the film (air side and plate side). The nature of the G/ATB film influenced the moisture and optical properties, while the films of G and G/MMT showed similar values. With regard to microstructure and roughness, the G film presented a more homogeneous and smooth surface, contrary to that was observed in films of G/MMT, who presented the highest roughness followed by G/ATB film. It was observed that there was a relationship between the roughness values and DF. According to the value of the CA, the three films had a hydrophobic character, because their values were above 65° (on both sides of the film), in the following order: G/MMT > G > G/ATB; the Cs followed the same trend. It should be also mentioned that a significant correlation between the values of CA and the roughness was found. According to the values of CA, Cs and SFE (specifically the polar component), the films of G/ATB had the lowest hydrophobicity, because had lower CA values and higher Cs values, as compared to the other two films. The values of the polar component of the film G/ATB were higher, which explains better the less hydrophobic character of this film. It can be concluded that the gelatin-based films produced in this work have hydrophobic properties (CA>65°), being the film G/MMT which showed better hydrophobic characteristics.
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Modelagem matemática do espalhamento do poluente mercúrio na águaConza, Adelaida Otazu January 2017 (has links)
O objetivo deste trabalho e a modelagem matem atica da propagaçãao do poluente mercúrio na agua. O modelo bidimensional consiste na drenagem da agua atrav es de um canal, onde o poluente (mercúrio) entra. O modelo consiste em um conjunto de equaçõoes diferenciais parciais: as equações para a conservação da massa, a quantidade de movimento, e a concentração das espécies, sujeitas a condições iniciais e de contorno apropriadas. Estas equações foram discretizadas pelo método de diferenças finitas centrais, gerando sistemas lineares que foram resolvidos pelo método de Gauss-Seidel e a convergência foi acelerada usando a técnica de sobre-relaxações SOR. A an alise da consistência e estabilidade da equação de concentração foi feita. Além disso, a solução analítica da equação de concentração, que e uma equação diferencial parcial bidimensional não homogênea com uma condição de contorno não homogênea, foi obtida com a transformada de Laplace. Os resultados obtidos a partir do modelo numérico e da solução analítica foram comparados e apresentam concordância razoável. / The goal of this work is the mathematical modeling of the spreading of the polluting mercury in the water. The two-dimensional model consists of water drainage through a canal, where the pollutant (mercury) enters. The model consists of a set of partial di erential equations: the equations for the conservation of the mass, the momentum, and the concentration of the species, subject to appropriate initial and boundary conditions. These equations were discretized by the method of central nite di erences, generating linear systems, which were solved by the Gauss-Seidel method and convergence was accelerated using the over-relaxation SOR technique. The analysis of the consistency and stability of the concentration equation was made. Furthermore, the analytical solution of the concentration equation, which is a two-dimensional non-homogeneous partial di erential equation with one nonhomogeneous contour condition, was obtained using Laplace transform. The results obtained from the numerical model and the analytical solution were compared and presented reasonable agreement.
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Modélisation de la mécanique de la cellule et son noyau dans le cadre de la migration confinée / Modeling cellular and nuclear mechanics in the context of confined migrationDeveraux, Solenne 11 September 2018 (has links)
Les cellules possèdent une capacitéfondamentale à leur survie : la migration. Del’embryogénèse aux métastases tumorales, lorsde la migration, les cellules doivent se faufiler àtravers des mailles sub-nucléaires pour atteindreleur localisation cible. Pour ce faire, ellespeuvent adapter leur mode locomotion ou leurspropriétés mécaniques à l’environnement quiles entoure. La cellule ainsi que son noyausubissent d’importantes déformations lors de lamigration en milieu confiné. Le noyau étantl’organelle le plus gros et le plus rigide, il peutlimiter la capacité migratoire de la cellule. Sespropriétés mécaniques sont donc décisives afinde migrer à travers un environnement complexe.Dans la littérature, les signaux moléculairespendant le processus migratoire ont étéabondamment décrits, mais la modélisationmécanique d’une cellule en migration peut-ellenous révéler de nouveaux éléments sur lesmécanismes sous-jacents ?La migration cellulaire est un procédé d’unecomplexité mécano-biologique telle, que tous sesaspects ne peuvent être modélisés à ce jour. Nousen choisissons donc trois que nousdévelopperons ici. Nous nous intéressonsd’abord à l’interaction mécanique entre le noyauet le cytoplasme lors d’une constriction de lacellule, puisque la plasticité du noyau sembleavoir un rôle primordial. Nous étudions ensuitele chimneying, un mode migratoire sansadhésion dont le mécanisme repose sur desforces de friction couplées à la poroélasticité ducytoplasme. Enfin, les substrats avec des micropilierssont depuis peu utilisés pour étudier lespropriétés mécaniques de la cellule et de sonnoyau, mais la mécanique de ce phénomène estpeu comprise. Nous modélisons le processus parlequel le noyau se déforme afin de déterminer s’ilest poussé ou tiré dans l’espace inter-piliers. / One of the fundamental properties incells is their ability to migrate. Fromembryogenesis to tumor metastasis, migratingcells must overcome mechanical obstacles toreach their intended location, squeezing throughsub-cellular and sub-nuclear gaps. It can be doneby adapting the locomotion mode to thesurrounding environment or by tuning the cell’sown mechanical properties. Migrating in aconfined space leads to intensive deformation ofthe cell and thus its nucleus. Being the largestand stiffest organelle, the nucleus can hamperthe migratory process. Its mechanical propertieshence are key to a successful migration in acomplex environment. Molecular signals behindcell migration have been extensively studied inthe literature, but what can computationalmechanics modeling unveil about themechanisms behind cell migration?Cell migration is such a complex mechanobiologicalprocess, that all aspects cannot bemodeled at once for now. We choose threedistinct situations for in-depth study. We firstseek to understand the mechanical interplaybetween the nucleus and the cytoplasm, sincenuclear plasticity seems decisive for migrationthrough sub-nuclear gaps. Second, weinvestigate the mechanics of chimneying, aspecific confined migratory mode, in which noadhesion in needed for the cell to move forward.Poroelasticity, coupled with friction, appears asthe key to successful locomotion. Eventually,cell spreading on micro-pillared substrates hasrecently been developed to study nuclearmechanical properties. The mechanism behindthis process being however unclear, we designeda large deformation model to determine whetherthe nucleus is being pushed or pulled in theinter-pillars gaps.
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Modelagem da dinâmica de doenças infecciosas em redes de movimentação de animais / Modeling the dynamics of infectious diseases in networks of animal movementsOssada, Raul 11 July 2011 (has links)
A dinâmica de movimentação de animais em uma rede de propriedades rurais e o espalhamento de algumas doenças animais estão intrinsecamente relacionados. Assim, compreender a dinâmica do espalhamento de doenças infecciosas nestas redes é um instrumento importante no controle destas. Neste projeto, foram implementados algoritmos para gerar redes de movimentação de animais hipotéticas e reconstruiu-se a rede de movimentações de bovinos do Estado do Mato Grosso, 2007, Brasil. Foram feitas diversas simulações a fim de verificar o espalhamento de doenças agudas e crônicas nessas redes. Diferentes dinâmicas de espalhamento de doenças infecciosas foram observadas em redes com a mesma distribuição de graus e diferentes estruturas topológicas. Espera-se que os resultados das simulações matemáticas possam auxiliar nas atividades dos órgãos responsáveis pela vigilância epidemiológica e incentivar outros Estados a seguirem o exemplo do Estado do Mato Grosso, a construírem bancos de dados que possam ser analisados utilizando a metodologia de redes. / The animals\' movements in a farms network and the spread of some animal diseases are intrinsically related. Therefore, comprehending the dynamics of the spreading of infectious diseases in these networks is an important tool in controlling these diseases. In this project, we have implemented algorithms to generate hypothetical networks of animals\' movements and rebuilt the network of bovine movements from the State of Mato Grosso, 2007, Brazil. We made several simulations in order to check the spreading of acute and chronic disease in these networks. Different dynamics of infectious disease spreading were observed in networks with the same degree distribution and different topological structure. We hope that the results of the mathematical simulations may assist in the activities of agencies responsible for disease surveillance and encourage other States to follow the example of the State of Mato Grosso, to build databases that can be analyzed using the methodology of networks.
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Multi-User Detection of Overloaded Systems with Low-Density SpreadingFantuz, Mitchell 11 September 2019 (has links)
Future wireless networks will have applications that require many devices to be connected to the network. Non-orthogonal multiple access (NOMA) is a promising multiple access scheme that allows more users to simultaneously transmit in a common channel than orthogonal signaling techniques. This overloading allows for high spectral efficiencies which can support the high demand for wireless access. One notable NOMA scheme is low-density spreading (LDS), which is a code domain multiple access scheme. Low density spreading operates like code division multiple access (CDMA) in the sense that users use a spreading sequence to spread their data, but the spreading sequences have a low number of nonzero chips, hence the term low-density. The message passing algorithm (MPA) is typically used for multi-user detection (MUD) of LDS systems. The MPA detector has complexity that is exponential to the number of users contributing to each chip. LDS systems suffer from two inherent problems: high computational complexity, and vulnerability to multipath channels. In this thesis, these two problems are addressed. A lower complexity MUD technique is presented, which offers complexity that is proportional to the number of users squared. The proposed detector is based on minimum mean square error (MMSE) and parallel interference cancellation (PIC) detectors. Simulation results show the proposed MUD technique achieves reductions in multiplications and additions by 81.84% and 67.87% with a loss of about 0.25 dB with overloading at 150%. In addition, a precoding scheme designed to mitigate the effects of the multipath channel is also presented. This precoding scheme applies an inverse channel response to the input signal before transmission. This allows for the received signal to eliminate the multipath effects that destroy the low-density structure.
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Engineered Surfaces for Biomaterials and Tissue EngineeringPeter George Unknown Date (has links)
The interaction of materials with biological systems is of critical importance to a vast number of applications from medical implants, tissue engineering scaffolds, blood-contacting devices, cell-culture products, as well as many other products in industries as diverse as agriculture. This thesis describes a method for the modification of biomaterial surfaces and the generation of tissue engineering scaffolds that utilises the self assembly of poly (styrene)-block-poly (ethylene oxide) (PS-PEO) block copolymers. Block copolymers consist of alternating segments of two or more chemically distinct polymers. The salient feature of these materials is their ability to self organise into a wide range of micro-phase separated structures generating patterned surfaces that have domain sizes in the order of 10-100nm. Further, it is also possible to specifically functionalise only one segment of the block copolymer, providing a means to precisely locate specific biological signals within the 10-100nm domains of a nano-patterned surface, formed via the programmed micro-phase separation of the block copolymer system. The density and spatial location of signalling molecules can be controlled by altering several variables, such as block length, block asymmetry, as well as processing parameters, providing the potential to authentically emulate the cellular micro to nano-environment and thus greatly improving on existing biomaterial and tissue engineering technologies. This thesis achieved several aims as outlined below; Developed methods to control the self-assembly of PS-PEO block copolymers and generate nano-patterned surfaces and scaffolds with utility for biomaterials applications. PS-PEO diblock copolymers were blended with polystyrene (PS) homopolymer and spin cast, resulting in the rapid self-assembly of vertically oriented PEO cylinders in a matrix of PS. Due to the kinetically constrained phase-separation of the system, increasing addition of homopolymer is shown to reduce the diameter of the PEO domains. This outcome provides a simple method that requires the adjustment of a single variable to tune the size of vertically oriented PEO domains between 10-100nm. Polymeric scaffolds for tissue engineering were manufactured via a method that combines macro-scale temperature induced phase separation with micro-phase separation of block copolymers. The phase behaviour of these polymer-solvent systems is described, and potential mechanisms leading to this spectacular structure formation are presented. The result is highly porous scaffolds with surfaces comprised of nano-scale self-assembled block copolymer domains, representing a significant advance in currently available technologies. Characterised the properties of these unique nano-structured materials as well as their interaction with proteinaceous fluids and cells. Nano-patterned PS-PEO self-assembled surfaces showed a significant reduction in protein adsorption compared to control PS surfaces. The adhesion of NIH 3T3 fibroblast cells was shown to be significantly affected by the surface coverage of PEO nano-domains formed by copolymer self-assembly. These nano-islands, when presented at high number density (almost 1000 domains per square micron), were shown to completely prevent cellular attachment, even though small amounts of protein were able to bind to the surface. In order to understand the mechanism by which these surfaces resisted protein and cellular adsorption we utilised neutron reflection to study their solvation and swelling properties. The results indicate that the PEO domains are highly solvated in water; however, the PEO chains do not extend into the solvent but remain in their isolated domains. The data supports growing evidence that the key mechanism by which PEO prevents protein adsorption is the blocking of protein adsorption sites. Control the nano-scale presentation of cellular adhesion and other biological molecules via the self-assembly of functionalised PS-PEO block copolymers Precise control over the nano-scale presentation of adhesion molecules and other biological factors represents a new frontier for biomaterials science. Recently, the control of integrin spacing and cellular shape has been shown to affect fundamental biological processes, including differentiation and apoptosis. We present the self-assembly of maleimide functionalised PS-PEO copolymers as a simple, yet highly precise method for controlling the position of cellular adhesion molecules. By controlling the phase separation of the functional PS-PEO block copolymer we alter the nano-scale (on PEO islands of 8-14 nm in size) presentation of the adhesion peptide, GRGDS, decreasing lateral spacing from 62 nm to 44 nm and increasing the number density from ~ 450 to ~ 900 islands per um2. The results indicate that the spreading of NIH-3T3 fibroblasts increases as the spacing between islands of RGD binding peptides decreases. Further, the same functional PS-PEO surfaces were utilised to immobilise poly-histidine tagged proteins and ECM fragments. The technologies developed in this thesis aim to improve on several weaknesses of existing biomaterials, in particular, directing cellular behaviour on surfaces, and within tissue engineering scaffolds, but also, on the prevention of fouling of biomaterials via non-specific protein adsorption. The application of block copolymer self-assembly for biomaterial and tissue engineering systems described in this thesis has great potential as a platform technology for the investigation of fundamental cell-surface and protein-surface interactions as well as for use in existing and emerging biomedical applications.
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Modelering av Wideband : Code Division Multiple Access / Behaviour modelling of Wideband : Code Division Multiple AccessHuynh, Jack, Gylin, Mattias January 2005 (has links)
<p>Today wireless transmission of data is becoming more and more popular and the need for faster transmission rates is increasing. Since the bandwidth is limited it is important to try to use it to the fullest. CDMA is a technology that allows multiple accesses on the same frequency and time thus making it very bandwidth efficient. The CDMA technology was first introduced in the second generation’s cellular systems but has since then been improved and is reused in today’s 3G systems as Wideband CDMA. ISY is interested in getting a behavioural model of a W-CDMA system since they had developed a DSP processor called BBP1 and were thinking about adding W-CDMA support for it. Even though our system is not implemented on the BBP1 it should provide a good base for future implementations. This thesis project will describe the construction of a behavioural model of a W-CDMA system following the standard specified by 3GPP. The system simulates W-CDMA transmission and reception and has an optional channel used to simulate real world interference. The receiver uses a rake combiner to improve the performance of the system.</p>
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Effects of invasin and YopH of Yersinia pseudotuberculosis on host cell signaling / Effekter av proteinerna invasin och YopH från bakterien Yersinia pseudotuberculosis på värdcellenGustavsson, Anna January 2004 (has links)
Integrins are a large family of membrane-spanning heterodimeric (αβ) receptors that bind to ligands on other cells or to extracellular matrix (ECM) proteins. These receptors mediate bidirectional signaling over the cell membrane to induce signaling cascades mediating functions as cell adhesion, spreading and migration. This signaling takes place at cell-matrix adhesions, which are sites where clustered and ligand-bound integrins connect to and mediate stabilization of the actin cytoskeleton, and induce signaling cascades. Integrins have a short cytoplasmic tail that is crucial for the bidirectional signaling, and the β1-integrin subunit exists in five splice variants only differing in the membrane-distal part of the cytoplasmic tail. This region of the almost ubiquitously expressed β1-integrin, β1A, contains two protein tyrosine motifs (NPXYs) interspaced with a threonine-rich region, while this region of the β1B splice variant is completely different and lacks known motifs. In contrast to the β1A-integrin, the β1B variant cannot mediate cell-matrix adhesion formation following binding to ECM ligands. The enteropathogenic bacterium Yersinia pseudotuberculosis binds to β1-integrins on the host cell with invasin, and this stimulates uptake of the bacterium. However, upon binding to the host cell, pathogenic Yersinia strains inject virulence effectors that block uptake. One effector responsible for the blocking is a tyrosine phosphatase, YopH. We identified the targets for this effector in the macrophage-like cell line J774A.1, which represent a professional phagocyte and thus is the likely target cell for the antiphagocytic effect of Yersinia. Two YopH target proteins were p130Cas and ADAP, of which the latter interestingly is an adapter protein specifically expressed in hematopoietic cells. ADAP has previously been implicated to participate in Fc-receptor-mediated phagocytosis and in communication between T-cell receptors and integrins. We also studied the importance of the cytoplasmic tail of β1-integrin for uptake of Yersinia. The GD25 cell line, which is a fibroblast-like cell line that lacks endogenous β1-integrins, was used together with GD25 cells transfected with β1B, β1Α or cytoplasmic tail mutants of β1A. These studies revealed that β1B-integrins could bind to invasin but not mediate uptake of Yersinia, while β1A both bound to invasin and mediated uptake. The first NPXY motif (unphosphorylated) and the double-threonines of the unique part of β1A were important for the ability of integrin to mediate uptake of Yersinia. These studies lead to the interesting finding that, when these cells were allowed to spread on invasin, those that expressed β1A spread as normal fibroblasts while for β1B-integrin-expressing cells, only finger-like protrusions of filopodia were formed. This provided us with a tool to study formation of filopodia without interference of the tightly linked process of lamellipodia formation. Initially, proteins that localized to the tip complex of these filopodia were identified. These were talin, VASP and interestingly the p130Cas-Crk-DOCK180 scaffold, while FAK, paxillin and vinculin were absent. In addition, VASP, p130Cas and Crk were shown to be important for the filopodia formation in GD25β1B. Further, the role of the actin motor myosin X, which previously has been implicated in formation of filopodia, was studied in the GD25Β1B cells and it was shown that myosin X not was important for filopodia formation, but that it recruited FAK and vinculin to the tip complexes of filopodia.
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