Les chondrocalcinoses articulaires (CCA) sont des arthropathies microcristallines caractérisées par la présence de cristaux de pyrophosphate de calcium dihydratés (PPCD) au sein du cartilage. Il existe trois formes de CCA : les formes familiales, transmises de façon autosomique dominante, et les formes sporadiques, dont la probabilité d'apparition augmente avec l'âge, et les formes secondaires à d'autres pathologies, comme l'hyperparathyroïdie. Le gène Ank code un transporteur exportant le pyrophosphate inorganique (PPi) hors des cellules. Un lien est clairement établi entre des mutations dans la séquence codante du gène Ank et les formes familiales de CCA Nous avons étudié la contribution du gène Ank dans la production de PPi par les chondrocytes articulaires stimulés avec du transforming growth factor-ß1 (TGF-ß1), leur sensibilité a ce facteur de croissance augmentant avec le l'âge des patients. Nous avons également, vu la présence fréquente des cristaux de PPCD chez les patients arthrosiques, et l'influence du niveau d'expression d'Ank dans la chondrogenèse, voulu élucider la possible implication d'Ank dans la perte du phénotype du chondrocyte articulaire, retrouvée dans l'arthrose. Enfin, nous avons étudié le rôle du calcium sur les effets du TGF-ß1 dans la production d'ePPi, l'hypercalcémie étant souvent retrouvée dans les tableaux cliniques d'hyperparathyroïdisme. Ces travaux ont permis de mettre en évidence le rôle majeur d'ANK en cas sensibilité accrue du chondrocyte au TGF-ß1. De plus, le PPi transporté par ANK influence le phénotype du chondrocyte articulaire. Enfin, le calcium joue un rôle crucial sur les effets inducteurs du TGF-ß1 dans la production de PPi. / Articular chondrocalcinosis (CCA) are microcrystalline arthtopathies characterized by the presence of calcium pyrophosphate dihydrate crystals (CPPD) in the cartilage. There are 3 forms of CCA, including the familal forms, dependent upon an autosomal dominant transmission mechanism, the sporadic forms, whose probability of occurrence increases with ageing, and the metabolic forms, secondary to other pathologies like hyperparathyroidism. The Ank gene encodes a transporter which exports inorganic pyrophosphate (PPi) outside the cells. A link is well established between mutations in the coding sequence of the Ank gene and the familial forms of CCA. We studied, on one hand, the contribution of the Ank gene in the production of PPi by articular chondrocytes stimulated with transforming growth factor-ß1 (TGF-ß1), since the sensitivity of chondrocytes to this growth factor increases with ageing. On the other hand, since CPPD are often associated with osteoarthritis (OA), and since Ank expression levels influences the chondrogenesis, we elucidated the possible implication of Ank in the loss of the articular chondrocytes phenotype observed during OA. Finally, we analyzed the role of calcium on the TGF-ß1 effects on ePPi production, hypercalcaemia being often found during hyperparathyroidism. This work allowed demonstrating, on one hand, the predominant role of ANK in the situations of chondrocytic hypersensitivity to TGF-ß1. On the other hand, the PPi exported by ANK influences the expression of articular chondrocytic markers. Finally, calcium plays a crucial role in the inducer effects of TGF-ß1 in the production of PPi.

The first objective of my work was to use the ank/ank (progressive ankylosis) mutant mice, which have a deficiency in inorganic pyrophosphate transport, to address the role of Ank in joint ankylosis. I observed the presence of hypertrophic chondrocytes in the uncalcified ank/ank mice articular cartilage. This novel phenotype is likely due to a dysregulation of chondrocyte maturation as these chondrocytes expressed hypertrophic chondrocyte markers (collagen type X and tissue non-specific alkaline phosphatase). I also showed by immunohistochemical staining that beta-catenin expression was upregulated and localized in the nuclei of articular ank/ank chondrocytes, suggesting activation of Wnt/beta-catenin signaling in these chondrocytes. The second objective was to use ank/ank mice as an informative model for understanding ankylosis mechanisms in human ankylosing spondylitis (AS) patients, as WNT/beta-catenin signaling plays an important role in ankylosis in AS patients. We attempted rescue of joint ankylosis in ank/ank mice by gene transfer of noggin, an antagonist of BMP signaling. Paradoxically, noggin-treated ank/ank mice had accelerated ankylosis, as evidenced by joint pathology and IHC staining of beta-catenin showed more intense signals in the spinal chondrocytes of the treated mice. As noggin and sclerostin (an antagonist of beta-catenin signaling) form a mutually inhibitory complex, we hypothesize that the formation of this complex results in relieving suppression of both beta-catenin and BMP signaling, leading to more severe ankylosis in ank/ank mice. By quantitative molecular imaging, I have demonstrated that ankylosis in these mutant mice developed simultaneously in distal and axial joint, instead of being a centripetal process. In summary, I have made three original observations in the ank/ank mice: the hypertrophic chondrocyte phenotype; activation of beta-catenin signaling and the simultaneous development of ankylosis in distal and axial joints. These mutant mice serve as valuable model for pre-clinical studies which enable modeling and testing of novel anti-ankylosis treatments.

The first objective of my work was to use the ank/ank (progressive ankylosis) mutant mice, which have a deficiency in inorganic pyrophosphate transport, to address the role of Ank in joint ankylosis. I observed the presence of hypertrophic chondrocytes in the uncalcified ank/ank mice articular cartilage. This novel phenotype is likely due to a dysregulation of chondrocyte maturation as these chondrocytes expressed hypertrophic chondrocyte markers (collagen type X and tissue non-specific alkaline phosphatase). I also showed by immunohistochemical staining that beta-catenin expression was upregulated and localized in the nuclei of articular ank/ank chondrocytes, suggesting activation of Wnt/beta-catenin signaling in these chondrocytes. The second objective was to use ank/ank mice as an informative model for understanding ankylosis mechanisms in human ankylosing spondylitis (AS) patients, as WNT/beta-catenin signaling plays an important role in ankylosis in AS patients. We attempted rescue of joint ankylosis in ank/ank mice by gene transfer of noggin, an antagonist of BMP signaling. Paradoxically, noggin-treated ank/ank mice had accelerated ankylosis, as evidenced by joint pathology and IHC staining of beta-catenin showed more intense signals in the spinal chondrocytes of the treated mice. As noggin and sclerostin (an antagonist of beta-catenin signaling) form a mutually inhibitory complex, we hypothesize that the formation of this complex results in relieving suppression of both beta-catenin and BMP signaling, leading to more severe ankylosis in ank/ank mice. By quantitative molecular imaging, I have demonstrated that ankylosis in these mutant mice developed simultaneously in distal and axial joint, instead of being a centripetal process. In summary, I have made three original observations in the ank/ank mice: the hypertrophic chondrocyte phenotype; activation of beta-catenin signaling and the simultaneous development of ankylosis in distal and axial joints. These mutant mice serve as valuable model for pre-clinical studies which enable modeling and testing of novel anti-ankylosis treatments.

En este trabajo de tesis doctoral se caracteriza funcional y molecularmente el canal TRPV4 en varios modelos de células epiteliales respiratorias mostrando por primera vez la participación de este canal en la función osmoreguladora a nivel celular así como la identificación de nuevas variantes del canal. Se demuestra que la entrada de Ca2+ en respuesta a un hinchamiento hipotónico se produce a través del canal TRPV4 y es necesaria para una eficiente recuperación del volumen o RVD. Por su parte, las células epiteliales respiratorias con fenotipo de fibrosis quística no son capaces de reducir su volumen en un medio hipotónico a causa de una regulación defectuosa del canal, indicando, al mismo tiempo, que la regulación del TRPV4 por el estímulo hipotónico es dependiente de la CFTR.La caracterización de las variantes del canal TRPV4 demuestra que los dominios de ANK son determinantes moleculares claves en el proceso de oligomerización del canal. Al mismo tiempo este trabajo describe nuevos aspectos relacionados con la biogénesis del TRPV4 hasta ahora desconocidos: la oligomerización del canal tiene lugar en el RE, orgánulo donde es N-glicosilado de forma simple antes de ser transportado hacia el Golgi donde sus N-glicanos son madurados. / This thesis characterizes molecularly and funcionally the TRPV4 channel in various models of airway epithelial cells showing, for the first time, the involvement of this channel in an osmoregulatory cellular function as well as the isolation of new splice variants of this channel. It is demonstrated that the TRPV4 channel is the molecular Ca2+ pathway activated by hypotonic estimulus needed to trigger the RVD response. Furthermore, the cystic fibrosis airway epithelial cells showed an impaired RVD due to the misregulation of the TRPV4 channel, indicating that the regulation by the hypotonic stimulus is CFTR-dependent.The characterization of the new variants demonstrated that the ANK domains are key structural determinants in the oligomerization process of the TRPV4. This work also describes new aspects related to the biogenesis of this channel: oligomerization is achieved in the ER, where the TRPV4 is N-glycosilated and then transported to the Golgi where the glycans are matured.

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2017-06-21T12:08:58Z No. of bitstreams: 2 Tese - Leonardo Martins de Abreu - 2017.pdf: 3441219 bytes, checksum: 28d75017e1ff1cdccf27de35d71771ea (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-07-10T13:08:28Z (GMT) No. of bitstreams: 2 Tese - Leonardo Martins de Abreu - 2017.pdf: 3441219 bytes, checksum: 28d75017e1ff1cdccf27de35d71771ea (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-07-10T13:08:28Z (GMT). No. of bitstreams: 2 Tese - Leonardo Martins de Abreu - 2017.pdf: 3441219 bytes, checksum: 28d75017e1ff1cdccf27de35d71771ea (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-05-12 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Using continuum and discrete solvent description models, we have investigated the chain length dependence of hydration free energy of polyol, glycine and ANK peptides. In this work this property values were calculated using thermodynamics integration with classical molecular dynamics simulation of one solute molecule to around 1000 solvent molecules. In addition, hydration free energy of polyol, glycine and ANK peptides were also calculated using polarizable continuum model in combination with the universal solvation model. Dipole moment and linear polarizability of the three systems had been also calculate using the PCM method. Our MD results show that ΔG exhibit a linear pattern with the size of glycine and ANK chain as well as observed for organic compounds in literature. In opposition, polyol’s ΔG shows a nonlinear behavior with increasing chain length and for this group of molecules it were found that water solvating effects have an important role over the polyol’s electric properties. These results suggest that in the case of polyols not only variation of polarizability but also variations of dipole moment in going from vacuum to water can influence the behavior of the free energy of hydration. All DM and PCM calculations were performed using the GROMACS 4.5 and the GAUSSIAN09 programs, respectively. / Utilizando modelos contínuo e discreto de descrição de solventes investigamos o comportamento da variação da energia livre na hidratação de series homólogas de poliol, glicina e peptídeos ANK. Neste trabalho os valores da energia de hidratação foram obtidos por integração termodinâmica utilizando simulações de dinâmica molecular clássica em sistemas com uma molécula de soluto para aproximadamente 1000 moléculas de solvente. Adicionalmente, o s resultados de ΔG foram obtidos utilizando o modelo contínuo polarizável combinado com o modelo de solvatação universal. Com o modelo PCM estudamos ainda o momento de dipolo e a polarizabilidade linear destes sistemas em questão e analisamos as contribuições que estas propriedades tem sobre a energia livre de hidratação e sua evolução com o crescimento de sistemas. Os resultados obtidos mostram que a variação da energia livre em função do número de resíduos cresce em valor absoluto para os três sistemas estudados. Para as moléculas de glicina e de peptídeos ANK observa-se um comportamento linear de ΔG com o número N de subunidades do sistema, semelhante ao observado para sistemas moleculares apresentados na literatura. Diferentemente, para as moléculas de poliol, os resultamos de ΔG exibem um comportamento não linear com o crescimento do sistema. Para esse grupo de moléculas inclusão do solvente tem grande impacto sobre o momento de dipolo e sobre a polarizabilidade linear. Este resultado mostra que para as moléculas de poliol não apenas a polarizabilidade, mas também variações no momento de dipolo podem influenciar o comportamento da energia livre de hidratação. Todos os cálculos foram realizados utilizando o programa GROMACS 4.5.4 e GAUSSIAN09.

This study aims to develop a method to investigate the relationship between settlement locations and the morphological landforms using geographical information systems (GIS). The method is applied to &Ccedil / ankiri province, a mountainous terrain, which is covered in seventy-seven sheets of topographic maps at 1:25.000 scale. Three databases are created and used in this study: 1) Settlement database comprising various topographic and landform attributes of 891 settlements, 2) Morphological landform database composed of 4042 landform polygon elements digitized from 1:25.000 topographic maps, and 3) Topographic database containing the digital elevation model of the area and its derivatives. The first step in the algorithm is to classify the area into four main landform classes, namely, valley, slope, flood and top. Unsuitable landforms are then clipped out based on the thresholds derived from three topographic properties (elevation, slope and aspect). Accordingly, about 2 % of the settlements and 12 % of the area are removed. The relationship is investigated using the percentages of remaining settlements and landform classes. Further analyses such as position of the settlement within the landform polygon and type of the nearest landforms are carried out for final interpretation. Following conclusions are reached on the relationship between settlement location and morphological landforms: - Percentages of settlements for flood, valley, slope and top are 8.37, 27.52, 58.60 and 5.50, respectively. Considering the percentages of the landforms provided in the area, however, valley is the most preferred landform followed by flood type. Slope and top landforms are less preferred. About 86 % of the settlements are concentrated along valley-slope boundary. - Morphological boundaries of flood and top landforms are consistent with the settlement zones. The valley-slope boundary, on the other hand, which is the most populated area, cross-cut the settlements zones.

Thermotherapy using natural thermal mud has played a crucial role for human health care since ancient times. However, recently the SPA and Wellness Centers have come into prominence with the exhaustion of primary reserves of natural thermal mud. In these centers, instead of natural thermal mud, peloids prepared using clay and/or clay minerals with thermal or mineral water through maturation process that changes the physicochemical properties of the clay and/or clay minerals. Turkey has several clay reserves / however, the investigation about their suitability for the pelotherapic applications is not adequate. The aim of this study is to investigate the mineralogical, chemical, and physicochemical properties of the 13 different samples collected from K&uuml / &ccedil / &uuml / khacibey, Hancili, and Kazbekir localities in &Ccedil / ankiri and Kizil&ouml / ren, Karadirek, and H&uuml / dai localities in Afyonkarahisar in order to evaluate their suitability for pelotherapic applications. The results of mineralogical, chemical, and physicochemical analyses including cation exchange capacity, swelling index, plastic index, water loss, specific surface area, grain size and cooling rate properties were compared with the results in the previous studies. For maturation process distilled water and two different commercial mineral water with different chemical compositions were used. At the end of the four week long maturation, the peloids are evaluated in the sense of consistency, ease of handling and pleasant sensation during application. Evaluation of analyses results showed that clay deposits in Hancili and K&uuml / &ccedil / &uuml / khacibey localities in &Ccedil / ankiri can be considered being suitable for pelotherapic applications. On the other hand, clay deposits in Kazbekir locality in &Ccedil / ankiri and in all three localities in Afyonkarahisar cannot be considered being suitable for pelotherapic purposes.

This study aims to develop a method to investigate the relationship between socio-economic status of village settlements and some topographic variables using geographical information systems (GIS) and spatial statistical methods. The study area is &Ccedil / ankiri province, a mountainous region that lays at the northeast of Ankara. 331 villages represented by areal units are used in this study. 195 variables are used to extract a single socio-economic status indicator. First, all the variables are divided under three groups, namely economic, social and service. Principal Components Analysis (PCA) is used to construct an index indicating socio-economic status. The parameters that represent natural environment are / mean elevation, mean slope, mean aspect and the ratio of high quality soil in the total area, for each settlement unit. The data is visualized by choropleth, cartogram and 3D techniques. Then it is explored by using correlograms, spatial moving averages and geographically weighted regression (GWR). Finally linear non-spatial regression and spatial regression methods are utilized in order to establish a relation between the socio-economic status and environmental parameters.

This study introduces an approach that seeks a possible relationship between settlement locations, water resources and rock types. The method is applied to &Ccedil / ankiri province (central-north Anatolia) which covers approximately an area of 8380 km2. Three main data sets used in this study. These are settlement, water and rock type data.The methodology of the study is composed of five steps. The first step is the conversion of all water data into a standardized point data. Total number of point data for water is 23911 after this step. The second step is to find the distances between water resources and settlements. In the third step the densities of water resources and settlements are derived and then tested for the rock types in particular areas. The fourth step is the overlay analysis in which all three data sets are combined to find preferred and avoided regions of settlements in relation to water resources and rock types. In the last step all analyses are integrated to extract information on effect of two parameters on the selection of a site. The main conclusions derived from the analysis are that: a) the mean and median distances between settlements and water resources are, 285 m and 163 m respectively, b) there is a strong relationship between water resources and settlement area, c) old clastics is the mostly preferred rock type whereas the carbonate rocks are mostly avoided.

No description available.