Orientador: Taís Maria Bauab / Banca: Sérgio Aparecido Torres / Banca: Daisy Nakamura Sato / Resumo: O Brasil apresenta uma grande diversidade de plantas medicinais, cuja utilização pela população vem aumentando cada vez mais. Byrsonima (Malpighiaceae) e Alchornea (Euphorbiaceae) que são plantas do Bioma-Cerrado do estado de São Paulo apresentam compostos com grande potencial de atividade biológica, sendo entretanto, pouco estudadas. A atividade antimicrobiana é uma etapa de extrema importância na caracterização biológica de produtos naturais com potencial farmacológico. Nossos objetivos foram investigar a atividade antibacteriana de extratos metanólicos e clorofórmicos de Byrsonima spp. e Alchornea spp empregando as técnicas de diluição em tubos e em microplacas com leitura espectrofotométrica e visual utilizando resazurina como revelador. Foram utilizadas as bactérias: Escherechia coli, Staphylococcus aureus, Yersinia enterocolitica e Aeromonas hydrophila. Nos ensaios em microplacas e em tubos foi determinada a concentração inibitória mínima (CIM) dos extratos vegetais testados à concentração de 1000æg/mL a 31,25æg/mL. Objetivou-se também comparar a eficácia das metodologias empregadas. Nosso estudo demonstrou maior atividade antibacteriana dos extratos metanólicos tanto de Byrsonima quanto de Alchornea, ressaltando os dados obtidos com B. crassa (MeOH) e B. basiloba (MeOH) que apresentaram acentuada atividade antibacteriana frente aos diferentes microrganismos testados. As metodologias empregadas foram eficazes para essa finalidade sendo as de diluição em microplacas mais vantajosas, pelo emprego de pequenos volumes principalmente os de extratos vegetais que são obtidos em pequenas quantidades. Dentre os dois métodos de microdiluição, a que empregava leitura espectrofotométrica apresentou-se mais sensível, porém a de leitura visual mostrou-se mais prática na leitura final e na interpretação dos resultados. / Abstract: Brazil presents a great diversity of medicinal plants, whose use by people increases more and more. Byrsonima (Malpighiaceae) and Alchornea (Euphorbiaceae) are plants of Bioma-Cerrado of the state of São Paulo and present composites with great potential of biological activity, but, however, they are little studied. The antimicrobial activity is a stage of extreme importance in the biological characterization of natural products with pharmacological potential. Our objectives were to investigate the antibacterial activity of methanolic and chlorophormic extracts of Byrsonima spp and Alchornea spp using the techniques of dilution in tubes and in microplates with spectrophotometric and visual readings using resazurin. Escherichia coli, Staphylococcus aureus, Yersinia enterocolitica and Aeromonas hydrophila were tested. In the microplate and tubes assays, the minimum innhibitory concentration (MIC) of plant extracts were tested in concentrations between 1000æg/mL and 31.25æg/mL. It was also compared the effectiveness of the methodologies employed. Our study demonstrated greater antibacterial activity of methanolic extracts of both Byrsonima and Alchornea, standing out the data obtained for B. crassa (MeOH) and B. basiloba (MeOH), which presented accented antibacterial activity against the different tested microrganisms. The methodologies employed were efficient for this purpose and the microplate dilution assay is more advantageous, for the fact it uses smaller volumes, mainly for those vegetal extracts that normally are obtained in small amounts. Amongst the two methods of microdilution, the one that used spectrophotometric reading proved to be more sensitive, however, the visual reading procedure was shown to be more practical in the final reading and interpretation of the results. / Mestre

O ensaio turbidimétrico tem como princípio a redução da densidade óptica em suspensões microbianas, decorrente o incremento na concentração do antibiótico. No ensaio convencional trabalha-se com tubos de ensaio que demandam longo tempo para visualização da resposta. Assim, o método empregando microplacas com leitura cinética para a dosagem de antibióticos é de interesse considerável, uma vez que possibilita reduzir quantidade de material e tempo de análise necessários e permite o ensaio de grande número de amostras simultaneamente, com leitura e cálculo automatizado. O objetivo deste trabalho é desenvolver o método de doseamento microbiológico de vancomicina empregando microplacas e modo de leitura cinético, assim como validar o método desenvolvido, através da avaliação dos parâmetros de especificidade e seletividade, linearidade, faixa ou intervalo linear, exatidão e precisão. Adicionalmente foi contemplado desenvolvimento paralelo entre o método convencional e o cinético, de forma a permitir respaldo positivo quanto à validação. Sendo assim, através do estudo dos parâmetros de validação do método cinético, trabalhou-se em condições estabelecidas abrangendo curva-padrão de vancomicina com concentrações entre 0,6 e 1,0 µg/mL, e emprego de meio de cultura caldo de caseína-soja inoculado com Bacillus subtilis (ATCC 6633) na proporção de 5%. Foram obtidos resultados satisfatórios em 5 horas de incubação. Conclui-se que o ensaio em microplacas permite avaliar, com vantagens, a atividade do antibiótico frente ao microrganismo-teste, permitindo facilidade operacional e maior rapidez na obtenção dos resultados. / The principle of the turbidimetric assay is to reduce the optical density of microbial suspensions, due to the increase in the concentration of antibiotic. In the conventional test works with tubes that require long time to display the answer. Thus, the method using microplates with kinetic reading for the dosage of antibiotics is of considerable interest, since it allows possible to reduce the amount of material and analysis time required and allows for testing of large numbers of samples simultaneously, with automated reading and calculating. The aim of this work is to develop the method of microbiological assay of vancomycin using microplates and kinetic reading mode, and to validate the method developed by evaluating the parameters of specificity and selectivity, linearity, linear range, accuracy and precision. Also included was the parallel development between the conventional method and the kinetic, to enable positive support for the validation. Thus, by studying the validation parameters of the kinetic method, worked under conditions laid down covering vancomycin standard curve with concentrations between 0.6 and 1.0 mg / mL, and soybean-casein broth inoculated with 5% of Bacillus subtilis (ATCC 6633). Satisfactory results were obtained with 5 hours incubation. In conclusion, the microplate assay allows to evaluate, with advantages, the activity of the antibiotic against the microorganism, allowing greater operational ease and rapidity of obtaining results.

The Maneadero Basin is identified as a transtensional sedimentary basin along the Agua Blanca Fault (ABF) in the southern limit to the "Big Bend" Domain of the North American-Pacific plate boundary zone. The ABF exhibits both the dextral and normal components of slip. This creates an interesting setting for the formation of the Maneadero Basin because structures with orientations similar to the ABF are typically contractional (e.g.: Puente Hills Fault, Whittier Fault, and Rancho Cucamonga Fault). The question if this basin is evidence of plate-scale transtension or local extension associated with bends/stopovers along the ABF is addressed by this study with three working hypotheses. The hypotheses presented by this study are: 1) the basins formed by a dip-slip component on the ABF and truly are an expression of regional transtension, 2) the basins formed at right steps along the dextral ABF, or 3) the basins formed as a result of juxtaposing basement blocks with disparate topographies. Each of these hypotheses would produce unique basin geometries and structures within and around the Maneadero Basin. To test these aforementioned hypotheses, a multi-disciplined study was conducted in the basin. A structural dataset was collected to identify kinematics and offsets of faults both within and bounding the basin. A gravity survey was also conducted to image the basin geometry. The results of the study show an asymmetrical gravity anomaly that closely follows the trace of the ABF. The amplitude of the anomaly is 54 mGal, the gradient of which is steepest around the ABF and shallows away from the fault to the north and east. Forward models of this anomaly indicate the ABF is a steeply north dipping fault. The gravity anomaly also indicates that the deepest part of the basin is located close to, but not coincident with the ABF and the basin gradually shallows to the northeast. This geometry is consistent with the hypothesis that the basin results from dip-slip on the ABF. This idea is also supported by the structural data, which includes fluvial terraces that have been uplifted and offset by faulting on the ABF, and by the presence of a normal fault on the ABF in the center of the basin. The third hypothesis is also supported by models of the gravity data, which suggest a deep (~900m) bowl shaped erosional feature in the bedrock. Dextral slip on the ABF juxtaposes the topographically high Punta Banda Ridge with this topographically low feature. Overall, the data presented in this study suggest the formation of the Maneadero Basin results from is a combination of the dip-slip component on the ABF and juxtaposition of the topographically elevated Punta Banda Ridge with a topographically lower basin of Bahia Todo Santos and Valle Maneadero. Geodetic data strongly suggest that the difference in motion of the Baja Microplate (south of the ABF) to the disrupted southern California Block (north of the ABF), and the orientation of the ABF relative to that motion, is causing transtension in the Maneadero Basin. This combined with strike-slip juxtaposition of different topographies allowed for the formation and evolution of the Maneadero Basin.

A FDA tem incentivado a indústria farmacêutica a implantar no seu sistema de controle de qualidade métodos rápidos para avaliação do produto durante seu processo de produção e/ou na sua forma final para comercialização. Na indústria farmacêutica, o doseamento microbiológico é amplamente utilizado pelo setor de Controle de Qualidade com objetivo de avaliar a eficácia e segurança do medicamento produzido, principalmente no que diz respeito a produção de antibióticos. No caso desses, a aplicabilidade de métodos rápidos é de suma importância a fim de se obter uma avaliação rápida e confiável do produto, garantindo assim a comercialização de medicamentos com a qualidade e segurança necessárias para o sucesso do tratamento terapêutico. O objetivo deste trabalho foi desenvolver e validar método para doseamento microbiológico de nistatina, com leitura cinética em microplaca, perfeitamente intercambiável com o método tradicional de difusão em ágar, porém com redução do tempo de execução e de liberação do resultado final. Os parâmetros definidos para este ensaio foram: utilização de inóculo com 5% de Candida albicans (ATCC 10231), meio de cultura líquido, tempos de 48 horas para crescimento do micro-organismo e 8 horas para leitura das micro-placas, , temperatura de análise de 30 °C e absorbância de 630 nM. Os parâmetros avaliados no processo de validação, de acordo com os códigos oficiais vigentes, demonstraram que o método é sensível, robusto, linear (r = 0,97332), precisão (repetibilidade: 96 %, DPR= 6 % / precisão intermediária: 97 %; DPR = 7 %) e exato ( 97 %, DPR = 7 %). Em comparação ao método de difusão em ágar tradicional, o método desenvolvido demonstrou a mesma eficiência do método já utilizado, contemplando todos os parâmetros do processo de validação, com a vantagem de ser realizado em menor tempo e com o uso reduzido de material de análise, como meio de cultura, padrão e amostra. Em função disso, pode-se concluir que o método desenvolvido e validado além de representar uma tendência mundial, por ser efetuado de modo rápido, eficaz e econômico, é perfeitamente aplicável a rotina laboratorial. / The FDA have been encouraging the pharmaceutical industry to deploy in their system of quality control a rapid method for assessment of the product during its production process and/or in their final form for sale. In the pharmaceutical industry, the microbiological assay is widely used by the Quality Control sector to evaluate the efficacy and safety of the product produced, especially as regards the production of antibiotics. In such case, the applicability of rapid methods is very important in order to obtain a rapid and reliable evaluation of the product, thus ensuring the marketing of medicinal products with the quality and security necessary for the success of therapeutic treatment. The aim of this study was to develop and validate an method for the microbiological assay of Nystatin with kinetic micro-plate reading, perfectly interchangeable with the traditional method of agar diffusion, but with reduced time for implementation and release of the final result. The parameters set for this assay were using 5 % inoculum of Candida albicans (ATCC 10231), liquid culture medium, time of 48 hours for growth of micro-organisms and 8 hours for reading the micro-plate temperature analysis of 30 °C and absorbance of 630 nM. The parameters evaluated in the validation process, according to the current official codes, shows that the method is sensitive, robust, linear (r = 0,97332), precision (repeatability: 96 %, RDS = 6 % / inter-day precision = 97 %, RSD = 7%) and accurate (97 %, RDS = 7%). Compared to the traditional method of diffusion in agar, the developed method demonstrated the same efficiency of the method already used, covering all parameters of the validation process, with the advantage of being performed in less time and with reduced use of material for analysis, as the culture medium and standard sample. As a result, it can be concluded that the method developed and validated besides representing a global trend, being made fast, effective and economical way, is perfectly applicable to routine clinical practice.

Orientador: Hélio José Montassier / Banca: Liana Brentano / Banca: Manoel Victor Franco Lemos / Resumo: A técnica de imunocaptura associada com a reação de transcrição reversa e reação em cadeia da polimerase (IC-RT-PCR) executadas tanto pelos procedimentos convencional como em tempo real foram testadas para a detecção rápida do gene da glicoproteína de Matriz (M) do vírus de influenza aviária (AIV) em amostras de líquido cório-alantóide (LCA) e em suabes traqueais e cloacais. O presente trabalho teve como objetivo desenvolver e otimizar a técnica de IC-RT-PCR para o diagnóstico do vírus da Influenza aviária. Os resultados obtidos foram comparados com um sistema empregando "beads" magnéticas em microplacas (AMBION), que é o método padrão de extração de RNA usado no laboratório de referência para diagnóstico de influenza aviária, o National Veterinary Services Laboratory - Ames, EUA (USDA), acrescido ainda de outros métodos de extração tradicionalmente usados nos laboratórios de referência para AIV, como os procedimentos com o uso do solvente orgânico Trizol® (Invitrogen) e com um sistema robotizado e que utiliza "beads" magnéticas (MagNA Pure - ROCHE). A técnica de IC-RT-PCR em tempo real neste estudo detectou a estirpe H2N2 do AIV, sem que nenhum outro dos RNA-vírus heterólogos testados fossem detectados (vírus das doença de Gumboro, de Newcastle e da bronquite infecciosa aviária). Os limites de detecção do IC-RTPCR foram iguais aos obtidos na técnica de extração com o kit da AMBION e menores do que aqueles que foram observados para os métodos de extração com Trizol® (Invitrogen) e com o MagNA Pure. O IC-RT-PCR demonstrou ser um sistema de diagnóstico capaz de conciliar simplicidade operacional e um menor custo com sensibilidade e especificidade analíticas iguais às do procedimento padrão atualmente adotado, podendo ser inclusive por laboratórios dotados de uma infra-estrutura mais simples / Abstract: The polymerase chain reaction (PCR) and reverse transcription-PCR (RT-PCR), including real-time RT-PCR have been used for the rapid detection of Matrix glycoprotein gene (M gene) Avian influenza virus (AIV). Despite the availability of various RNA extraction methods for using in RT-PCR, isolation and detection of viral RNA are still difficult due to the unstable nature of viral RNA molecules and the presence of PCR inhibitory substances. In this study, a simple method using immune-capture (IC) to recover viral RNA from H2 AIV samples was developed and compared to one standard and two others reference methods used for viral RNA extraction, such as Ambion MagMAXTM kit and Trizol® (Invitrogen) and Magnapure kit (Roche), respectively, with subsequent analysis by real-time RT-PCR. The real-time IC-RT-PCR developed in was able to detect specifically H2N2 AIV strain, without detecting non-related avian RNA-virus pathogens, such as Newcastle disease virus, avian infectious bronchitis virus and Gumboro disease virus. Comparable detection limits were found for IC and the standard RNA extraction method using Ambion MagMAXTM kit, either for the detection of AIV in allantoic fluid suspension or in seeded tracheal and cloacal swab samples by conventional or real time RT-PCR techniques. These methods were less sensitive than Trizol® (Invitrogen) and Magnapure kit (Roche) procedures. Thus, IC was rapid and as sensitive and specific as current standard AIV RNA extraction method for real time or conventional RT-PCR, besides it conciliated simplicity and lower cost and can be applied simultaneously for direct detection of AIV in a large number of samples, including less-equipped laboratories / Mestre

Made available in DSpace on 2014-06-11T19:27:23Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-08-13Bitstream added on 2014-06-13T20:16:39Z : No. of bitstreams: 1 dipillo_f_me_jabo.pdf: 266438 bytes, checksum: 248a318a5e5422d95006058e8dd1370a (MD5) / A técnica de imunocaptura associada com a reação de transcrição reversa e reação em cadeia da polimerase (IC-RT-PCR) executadas tanto pelos procedimentos convencional como em tempo real foram testadas para a detecção rápida do gene da glicoproteína de Matriz (M) do vírus de influenza aviária (AIV) em amostras de líquido cório-alantóide (LCA) e em suabes traqueais e cloacais. O presente trabalho teve como objetivo desenvolver e otimizar a técnica de IC-RT-PCR para o diagnóstico do vírus da Influenza aviária. Os resultados obtidos foram comparados com um sistema empregando “beads” magnéticas em microplacas (AMBION), que é o método padrão de extração de RNA usado no laboratório de referência para diagnóstico de influenza aviária, o National Veterinary Services Laboratory – Ames, EUA (USDA), acrescido ainda de outros métodos de extração tradicionalmente usados nos laboratórios de referência para AIV, como os procedimentos com o uso do solvente orgânico Trizol® (Invitrogen) e com um sistema robotizado e que utiliza “beads” magnéticas (MagNA Pure - ROCHE). A técnica de IC-RT-PCR em tempo real neste estudo detectou a estirpe H2N2 do AIV, sem que nenhum outro dos RNA-vírus heterólogos testados fossem detectados (vírus das doença de Gumboro, de Newcastle e da bronquite infecciosa aviária). Os limites de detecção do IC-RTPCR foram iguais aos obtidos na técnica de extração com o kit da AMBION e menores do que aqueles que foram observados para os métodos de extração com Trizol® (Invitrogen) e com o MagNA Pure. O IC-RT-PCR demonstrou ser um sistema de diagnóstico capaz de conciliar simplicidade operacional e um menor custo com sensibilidade e especificidade analíticas iguais às do procedimento padrão atualmente adotado, podendo ser inclusive por laboratórios dotados de uma infra-estrutura mais simples / The polymerase chain reaction (PCR) and reverse transcription-PCR (RT-PCR), including real-time RT-PCR have been used for the rapid detection of Matrix glycoprotein gene (M gene) Avian influenza virus (AIV). Despite the availability of various RNA extraction methods for using in RT-PCR, isolation and detection of viral RNA are still difficult due to the unstable nature of viral RNA molecules and the presence of PCR inhibitory substances. In this study, a simple method using immune-capture (IC) to recover viral RNA from H2 AIV samples was developed and compared to one standard and two others reference methods used for viral RNA extraction, such as Ambion MagMAXTM kit and Trizol® (Invitrogen) and Magnapure kit (Roche), respectively, with subsequent analysis by real-time RT-PCR. The real-time IC-RT-PCR developed in was able to detect specifically H2N2 AIV strain, without detecting non-related avian RNA-virus pathogens, such as Newcastle disease virus, avian infectious bronchitis virus and Gumboro disease virus. Comparable detection limits were found for IC and the standard RNA extraction method using Ambion MagMAXTM kit, either for the detection of AIV in allantoic fluid suspension or in seeded tracheal and cloacal swab samples by conventional or real time RT-PCR techniques. These methods were less sensitive than Trizol® (Invitrogen) and Magnapure kit (Roche) procedures. Thus, IC was rapid and as sensitive and specific as current standard AIV RNA extraction method for real time or conventional RT-PCR, besides it conciliated simplicity and lower cost and can be applied simultaneously for direct detection of AIV in a large number of samples, including less-equipped laboratories

A FDA tem incentivado a indústria farmacêutica a implantar no seu sistema de controle de qualidade métodos rápidos para avaliação do produto durante seu processo de produção e/ou na sua forma final para comercialização. Na indústria farmacêutica, o doseamento microbiológico é amplamente utilizado pelo setor de Controle de Qualidade com objetivo de avaliar a eficácia e segurança do medicamento produzido, principalmente no que diz respeito a produção de antibióticos. No caso desses, a aplicabilidade de métodos rápidos é de suma importância a fim de se obter uma avaliação rápida e confiável do produto, garantindo assim a comercialização de medicamentos com a qualidade e segurança necessárias para o sucesso do tratamento terapêutico. O objetivo deste trabalho foi desenvolver e validar método para doseamento microbiológico de nistatina, com leitura cinética em microplaca, perfeitamente intercambiável com o método tradicional de difusão em ágar, porém com redução do tempo de execução e de liberação do resultado final. Os parâmetros definidos para este ensaio foram: utilização de inóculo com 5% de Candida albicans (ATCC 10231), meio de cultura líquido, tempos de 48 horas para crescimento do micro-organismo e 8 horas para leitura das micro-placas, , temperatura de análise de 30 °C e absorbância de 630 nM. Os parâmetros avaliados no processo de validação, de acordo com os códigos oficiais vigentes, demonstraram que o método é sensível, robusto, linear (r = 0,97332), precisão (repetibilidade: 96 %, DPR= 6 % / precisão intermediária: 97 %; DPR = 7 %) e exato ( 97 %, DPR = 7 %). Em comparação ao método de difusão em ágar tradicional, o método desenvolvido demonstrou a mesma eficiência do método já utilizado, contemplando todos os parâmetros do processo de validação, com a vantagem de ser realizado em menor tempo e com o uso reduzido de material de análise, como meio de cultura, padrão e amostra. Em função disso, pode-se concluir que o método desenvolvido e validado além de representar uma tendência mundial, por ser efetuado de modo rápido, eficaz e econômico, é perfeitamente aplicável a rotina laboratorial. / The FDA have been encouraging the pharmaceutical industry to deploy in their system of quality control a rapid method for assessment of the product during its production process and/or in their final form for sale. In the pharmaceutical industry, the microbiological assay is widely used by the Quality Control sector to evaluate the efficacy and safety of the product produced, especially as regards the production of antibiotics. In such case, the applicability of rapid methods is very important in order to obtain a rapid and reliable evaluation of the product, thus ensuring the marketing of medicinal products with the quality and security necessary for the success of therapeutic treatment. The aim of this study was to develop and validate an method for the microbiological assay of Nystatin with kinetic micro-plate reading, perfectly interchangeable with the traditional method of agar diffusion, but with reduced time for implementation and release of the final result. The parameters set for this assay were using 5 % inoculum of Candida albicans (ATCC 10231), liquid culture medium, time of 48 hours for growth of micro-organisms and 8 hours for reading the micro-plate temperature analysis of 30 °C and absorbance of 630 nM. The parameters evaluated in the validation process, according to the current official codes, shows that the method is sensitive, robust, linear (r = 0,97332), precision (repeatability: 96 %, RDS = 6 % / inter-day precision = 97 %, RSD = 7%) and accurate (97 %, RDS = 7%). Compared to the traditional method of diffusion in agar, the developed method demonstrated the same efficiency of the method already used, covering all parameters of the validation process, with the advantage of being performed in less time and with reduced use of material for analysis, as the culture medium and standard sample. As a result, it can be concluded that the method developed and validated besides representing a global trend, being made fast, effective and economical way, is perfectly applicable to routine clinical practice.

Au Nord-Est de la microplaque Adriatique la jonction Alpes-Dinarides représente une région clé pour comprendre les interactions entre la microplaque et l’Europe stable. Alors que la tectonique active de la partie alpine de cette zone est relativement bien contrainte, peu de données sont disponibles quant aux déformations actives contrôlées par la rotation de la microplaque à travers les Dinarides. Par une approche morpho-tectonique (étude de terrain combinée à l’analyse d’images aériennes et satellitaires, de cartes topographiques et modèles numériques de terrain haute résolution) nous avons cartographié en détails les failles actives des Dinarides septentrionales et de la partie orientale des Alpes du Sud. Sur la base de cette cartographie et des données géologiques une quarantaine de décalages tectoniques cumulés allant de quelques mètres à plusieurs kilomètres a été identifiée. A l’aide de datations 36Cl de marqueurs morphologiques affectés par les failles combinées aux chronologies existantes les vitesses de déformation actuelles ont ensuite été estimées. L’évolution des déformations depuis le Pliocène a pu être contrainte dans les Dinarides mettant en évidence une initiation des failles au début du Pliocène et un changement cinématique important autour du Pléistocène moyen. Les vitesses obtenues, notamment environ 3mm/an de mouvement dextre le long des Dinarides, ont finalement été confrontées aux modèles existants ce qui a conduit à proposer un modèle cinématique décrivant l’accommodation de la rotation de l’Adriatique par le mouvement relatif de blocs lithosphériques rigides et qui permet d’expliquer les déformations actives observées aux frontières de ces blocs. / At the northeastern corner of the Adria microplate the Alps-Dinarides junction represents a key region to understand the interactions between the microplate and the stable Europe. While the active tectonics of the alpine part of the area is relatively well-known, few data allow characterizing the present-day deformations controlled by the microplate rotation across the Dinarides. Using a morpho-tectonic approach (field study combined to the analysis of aerial and satellite images, topographical maps and high-resolution digital elevation models) we mapped in details the active faults in the Northern Dinarides and the eastern part of the Southern Alps. Based on this mapping and geologic data forty tectonic cumulative displacements ranging from few meters to several kilometers have been identified. By determining the 36Cl exposure ages of faulted geomorphic markers and comparing it to existing chronologies the present-day rates of deformation have then been assessed. The evolution of the deformations since the Pliocene could also have been constrained revealing an Early Pliocene age for the onset of strike-slip faulting and a major kinematic change during the Middle Pleistocene. Finally the yielded faults slip-rates, especially about 3 mm/yr of right-lateral motion across the Dinarides, have been compared to existing models. That allowed proposing a kinematic model describing the Adria rotation accommodation through the relative motion of rigid lithospheric blocks and explaining the observed active deformations at their boundaries.

The goal of the current study was to develop a computational framework for modelling the coupled fluid-structure interaction problem of squeeze films often encountered in MEMS devices. Vibratory MEMS devices such as gyroscopes, RF switches, and 2D resonators often have a thin plate like structure vibrating transversely to a Fixed substrate, and are generally not perfectly vacuum packed. This results in a thin air film being trapped between the vibrating plate and the fixed substrate which behaves like a squeeze film offering both stiffness and damping to the vibrating plate. For accurate modelling of the squeeze film effect, one must account for the coupled fluid-structure interaction. The majority of prior works attempting to address the coupled problem either approximate the mode shape of the vibrating plate or resort to cumbersome iterative solution strategies to address the problem in an indirect way. In the current work, we discuss the development of a fully coupled finite element based numerical scheme to solve the 2D Reynolds equation coupled with the 3D plate elasticity equation in a single step. The squeeze film solver so developed has been implemented into a commercial FEA package NISA as part of its Micro-Systems module. Further, extending on a prior analytical work, the effect of variable ow boundaries for an all sides clamped plate on squeeze film parameters has been thoroughly investigated. The developed FEM based numerical scheme has been used to validate the results of the prior analytical study. The developed numerical scheme models the 2D Reynolds equation thus limiting the model to account for the effects of the fluid volume strictly confined between the structure and the substrate. To study the effect of surrounding fluid volume ANSYS FLOTRAN simulations have been performed by numerically solving the full 3D Navier Stokes equation in the extended fluid domain for the different flow boundary scenarios. Cut-off frequencies are established beyond which one can consider a 2D fluid domain without considerable loss of accuracy. First, a displacement based finite element formulation is presented for the 2D Reynolds equation coupled with the 3D elasticity equation. Both lower order 8 node and higher order 27 node 3D elements are developed. Only a single type of 3D element is used for modelling along with a 2D fluid layer represented by the \wet" face of the 3D structural domain. The results from our numerical model are compared with experimental data from literature for a MEMS cantilever. The results from the 27 node displacement based elements show good agreement with published experimental data. The results from the lower order 8 node displacement based elements however show huge errors even for relatively fine meshes due to locking issues in modelling high aspect ratio structures. This limits the implementation of the displacement based solver in commercial FE packages where the available mesh generators are generally restricted to lower order 3D elements. In order to overcome the limitations faced by lower order elements (primarily locking issues) in modelling high aspect ratio MEMS geometries, a coupled hybrid formulation is developed next. A thorough performance study is presented considering both the hybrid and displacement based elements for lower order 8 node and higher order 27 node ele- ments. The optimal element choice for modelling squeeze film geometries is determined based on the comparative studies. The effect of element aspect ratio for hybrid and displacement based elements are studied and the superiority of hybrid formulation over displacement based formulations is established for lower order 8 node elements. The coupled hybrid nite element formulation developed for lower order elements is implemented in the commercial FEA package NISA. The implementation scheme to integrate the developed coupled hybrid 8 node squeeze film solver into the commercial FEA package is discussed. The pre-integration analysis and subsequent requirement gaps are first investigated. Based on the gap analysis, certain GUI modifications are undertaken and parser programs are developed to re-format data according to NISA input requirements. Certain special features are included in the package to aid in post processing data analysis by MEMS designers such as \frequency sweep" and \node of interest" selection. As a case study for validation, we also present the modelling of a MEMS cantilever and show that the simulation results from our software are in good agreement with experimental data reported in the literature. Finally as a case study, an extension of a prior analytical work, which studies the effect of varying flow boundaries on squeeze film parameters, is discussed. Explanations are provided for the findings reported in the prior analytical work. The concept of using variation in flow boundaries as a frequency tuning tool is introduced. The analytical results are validated with the coupled numerical scheme discussed before, by considering imposed mode shape for an all sides clamped plate as prescribed displacement to the fluid domain. The simulated results are used to study the intricacies in squeeze film damping and stiffness variations with respect to spatial changes in the fluid flow boundary conditions. In particular, it has been shown that the boundary venting conditions can be used effectively to tune the dynamic response of a micromechanical structure over a fairly large range of frequencies and somewhat smaller range of squeeze film damping. Next, the effect of the surrounding fluid volume for various venting conditions is studied. ANSYS FLOTRAN is used to solve for the full 3D Navier Stokes equation over the extended fluid domain. Results from the extended domain study are used to determine cut-off frequencies beyond which one need not resort to an extended mesh study, and yet be within 5% accuracy of the full extended mesh model.

The thesis deals with the processing and detection of specific objects in the image on the Android mobile platform. The main objective of this work was to design and then implement a mobile application for Android operating system, which allows control of pipetting processes based on images from a mobile device camera. The OpenCV library is used in the application for image processing. The resulting application should serve primarily in laboratories as a tool for complete analysis of the pipetting process. The work is divided into two main chapters, which further consist of sections and smaller subsections. The first chapter is devoted to the theoretical analysis of this work. Here is also describes used technology, Android operating system, OpenCV library and important parts of image processing. The second chapter deals with the proposal and subsequent practical solution of this work. There is a proposal and the following procedure for solving this work, important techniques, methods of processing and analysis of the camera image. The conclusion of the thesis is an evaluation of the results of the solution of this work.