A significant body of literature has reported strategies and techniques to assess the mechanical properties of biological samples such as proteins, cellular and tissue systems. Atomic force microscopy has been used to detect elasticity changes of cancer cells. However, only a few studies have provided a detailed and complete protocol of the experimental procedures and data analysis methods for non-adherent blood cancer cells. In this work, the elasticity of NB4 cells derived from acute promyelocytic leukemia (APL) was probed by AFM indentation measurements to investigate the effects of the disease on cellular biomechanics. Understanding how leukemia influences the nanomechanical properties of cells is expected to provide a better understanding of the cellular mechanisms associated to cancer, and promises to become a valuable new tool for cancer detection and staging. In this context, the quantification of the mechanical properties of APL cells requires a systematic and optimized approach for data collection and analysis, in order to generate reproducible and comparative data. This Thesis elucidates the automated data analysis process that integrates programming, force curve collection and analysis optimization to assess variations of cell elasticity in response to processing criteria. A processing algorithm was developed by using the IGOR Pro software to automatically analyze large numbers of AFM data sets in an efficient and accurate manner. In fact, since the analysis involves multiple steps that must be repeated for many individual cells, an automated and un-biased processing approach is essential to precisely determine cell elasticity. Different fitting models for extracting the Young’s modulus have been systematically applied to validate the process, and the best fitting criteria, such as the contact point location and indentation length, have been determined in order to obtain consistent results. The designed automated processing code described in this Thesis was used to correlate alterations in cellular biomechanics of cancer cells as they undergo drug treatments. In order to fully assess drug effects on NB4 cells, viability assays were first performed using Trypan Blue staining for primary insights before initiating thorough microplate fluorescence intensity readings using a LIVE/DEAD viability kit involving ethidium and calcein AM labelling components. From 0 to 24 h after treatment using 30 µM arsenic trioxide, relative live cell populations increased until 36 h. From 0 to 12 h post-treatment, relative populations of dead cells increased until 24 h post-treatment. Furthermore, a drastic drop in dead cell count has been observed between 12 and 24 h. Additionally, arsenic trioxide drug induced alterations in elasticity of NB4 cells can be correlated to the cell viability tests. With respect to cell mechanics, trapping of the non-adherent NB4 cells within fabricated SU8-10 microwell arrays, allowed consistent AFM indentation measurements up to 48 h after treatment. Results revealed an increase in cell elasticity up to 12 h post-treatment and a drastic decrease between 12 and 24 h. Furthermore, arsenic trioxide drug induced alterations in elasticity of NB4 cells can be correlated to the cell viability tests. In addition to these indentation and viability testing approaches, morphological appearances were monitored, in order to track the apoptosis process of the affected cells. Relationships found between viability and elasticity assays in conjunction with morphology alterations revealed distinguish stages of apoptosis throughout treatment. 24 h after initial treatment, most cells were observed to have burst or displayed obvious blebbing. These relations between different measurement methods may reveal a potential drug screening approach, for understanding specific physical and biological of drug effects on the cancer cells.

INTRODUÇÃO: O envelhecimento da população é uma tendência social conhecida em países desenvolvidos e cada vez mais pronunciada em países em desenvolvimento. A demência é considerada um dos principais problemas de saúde devido ao rápido crescimento populacional de idosos, sendo os distúrbios de linguagem considerados importantes nesses quadros. O discurso tem ganhado destaque para a identificação dos distúrbios linguísticos nas demências assim como no seguimento desses pacientes. A caracterização das diferenças pode auxiliar no diagnóstico diferencial e contribuir para a criação de ferramentas futuras que auxiliem na intervenção clínica e ajudem a evitar a evolução e/ou progressão dos quadros demenciais. O processo de transcrição e análise do discurso é bastante laborioso, desta forma o uso de métodos computacionais tem auxiliado na identificação e extração de características linguísticas. OBJETIVO: identificar alterações em aspectos micro e macrolinguísticos que diferenciem indivíduos com doença de Alzheimer, comprometimento cognitivo leve e idosos sem comprometimento cognitivo na tarefa de narrativa de figuras em sequência e explorar a ferramenta computacional (Coh-Metrix-Dementia) para análise do discurso desses sujeitos. MÉTODO: Foram avaliados 60 indivíduos, sendo 20 em cada grupo de pesquisa (doença de Alzheimer leve - GDA, comprometimento cognitivo leve amnéstico - GCCLa e controle - GC). Os indivíduos foram solicitados a enunciar uma narrativa baseada em 22 cenas em sequência, que retratam a história da \"Cinderela\". Foram aplicados também os seguintes testes linguístico-cognitivos: Fluência Verbal, Teste de Nomeação do Boston e Camel and Cactus test. Utilizou-se o Coh-Metrix- Dementia para extração automática das métricas. RESULTADOS: Os valores extraídos pelo Coh-Metrix-Dementia foram tratados estatisticamente sendo possível levantar métricas capazes de distinguir os grupos estudados. Em relação aos aspectos microlinguísticos destacaram-se a redução nas habilidades sintáticas, maior dificuldade no resgate verbal, discursos com menor coesão e coerência local no GDA. No nível macrolinguístico o GDA apresentou os discursos menos informativos, com maior prejuízo em relação a coerência global e maior número de modalizações. O GDA também apresentou maior comprometimento da estrutura narrativa. Não foi possível discriminar o GCCLa e GC em nenhuma métrica do discurso deste estudo. Foram feitas adaptações em relação a segmentação das sentenças para um melhor funcionamento da ferramenta computacional. CONCLUSÃO: Os indivíduos do GDA apresentaram discursos com maior comprometimento macro e microestrutural. O uso da ferramenta computacional se mostrou um importante aliado para análises discursivas / INTRODUCTION: Population aging is a social trend known in developed countries and increasingly pronounced in developing countries. Dementia is considered one of the main health problems due to the rapid population growth of the elderly, and language disorders are considered important in these settings. The discourse is important for the identification of linguistic disorders in dementias as well as in the follow-up of these patients. The discourse differences characterization can help on the differential diagnosis and contribute to the creation of future tools for clinical intervention and help prevent the evolution and/or progression of dementia. The transcription and discourse analysis are laborius, thus the use of computational methods helped in the identification and extraction of linguistic characteristics. OBJECTIVE: The objective of this study was to identify changes in micro and macrolinguistic aspects that differentiate individuals with Alzheimer\'s disease, mild cognitive impairment and healthy elderly individuals during narrative of figures in sequence and to explore the computational tool (Coh-Metrix-Dementia) to analyze the subjects\' discourse. METHODS: 60 subjects were evaluated, 20 of them in each research group (mild Alzheimer\'s disease - GDA, amnestic cognitive impairment - GCCLa and control - CG). The subjects were asked to construct a narrative based on sequence of pictures, about the \"Cinderella´s Story\". The following linguistic-cognitive tests were also applied: Verbal Fluency, Boston Naming Test, and Camel and Cactus test. Coh-Metrix-Dementia was used for automatic metrics extraction. RESULTS: The values extracted by Coh-Metrix-Dementia were statistically treated and it was possible to obtain metrics capable of distinguishing the studied groups. In relation to the microlinguistic aspects, it was found the reduction in syntactic abilities, greater difficulty in verbal rescue, discourses with less cohesion and local coherence in the GDA. In the macrolinguistic level the GDA presented the less informative discourses, with greater loss in global coherence and the greater number of modalizations. The GDA also presented greater impairment on narrative structure. It was not possible to discriminate GCCLa and GC in any discourse´s metric in this study tool functioning. CONCLUSION: The GDA subjects presented discourses with greater macro and microstructural impairment. The computational tool usage proved to be an important ally for discursive analysis

INTRODUÇÃO: O tema de investigação, discurso de afásicos, fornece informações importantes sobre aspectos fonológicos, morfológicos, sintáticos, semânticos e pragmáticos da linguagem de pacientes que sofreram lesão vascular cerebral. Uma das maneiras de estudar o discurso é por meio de cenas figurativas temáticas simples ou em sequência. A sequência da história de \"Cinderela\" é frequentemente utilizada em estudos, por ser familiar em todo o mundo, o que favorece estudos transculturais; por induzir a produção de narrativas, ao invés de descrições, frequentemente obtidas quando se utiliza prancha única para eliciar discursos. Outra vantagem do uso das sequências da \"Cinderela\" é o fato de gerar material linguístico em quantidade suficiente para análise detalhada. OBJETIVOS: (1) analisar, por meio de tecnologias computadorizadas, aspectos macro e microestruturais do discurso de indivíduos sadios do ponto de vista cognitivo, afásicos de Broca e afásicos anômicos; (2) explorar o discurso como indicador de evolução da afasia; (3) analisar a contribuição do SPECT para verificação de evolução da afasia junto ao discurso. MÉTODO: Participaram do estudo oito indivíduos afásicos de Broca e anômicos que compuseram o grupo do estudo longitudinal (G1), 15 indivíduos afásicos de Broca e anômicos que compuseram o outro grupo de estudo (G2) e 30 cognitivamente sadios (GC). Os participantes foram solicitados a examinar as cenas da história \"Cinderela\" e depois recontar a história, com suas palavras. Foram exploradas tecnologias computadorizadas e analisados aspectos macro e microestruturais dos discursos produzidos. Para o G1, tivermos a particularidade de coleta de discurso também pela prancha \"Roubo dos Biscoitos\", análise do exame SPECT e acompanhamento longitudinal por um período de seis meses. RESULTADOS: Comparando o GC e o G2, em relação à macroestrutura, notou-se que os afásicos do G2 se diferenciaram significativamente do GC em todas as proposições e, em relação à microestrutura, sete métricas foram capazes de diferenciar ambos os grupos. Houve diferença significante macro e micro estrutural entre os sujeitos afásicos de Broca e anômicos. Foi possível verificar diferenças em medidas da macro e da microestrutura no G1 com o avançar do tempo de lesão após AVC. A história da \"Cinderela\" forneceu dados de microestrutura mais completos do que a prancha \"Roubo dos Biscoitos\". Os resultados do SPECT permaneceram os mesmos, sem demonstração de mudança com a evolução da afasia. CONCLUSÃO: A produção de narrativa gerou material para análise de macroestrutura e microestrutura, tanto aspectos de macro quanto de microestrutura diferenciaram indivíduos cognitivamente sadios dos sujeitos afásicos. A análise do discurso da \"Cinderela\" serviu como instrumento para mensurar a melhora da linguagem dos sujeitos afásicos. O uso da ferramenta computacional auxiliou as análises discursivas / INTRODUCTION: The aphasic discourse analysis provides important information about the phonological, morphological, syntactic, semantic and pragmatic aspects of the language of patients who have suffered a stroke. The evaluation of the discourse, along with other methods, can contribute to observation of the evolution of the language and communication of aphasic patients; however, manual analysis is laborious and can lead to errors. OBJECTIVES: (1) to analyze, by computerized technologies, macro and microstructural aspects of the discourse of healthy cognitive individuals, Broca\'s and anomic aphasics; (2) to explore the discourse as indicator of the evolution of aphasia; (3) to analyze the contribution of single photon emission computed tomography (SPECT) to verify the correlation between behavioral and neuroimaging evolution data. METHOD: Two groups of patients were studied: GA1, consisting of eight individuals with Broca\'s aphasia and anomic aphasia, who were analyzed longitudinally from the sub-acute phase of the lesion and after three and six months; GA2 composed of 15 individuals with Broca\'s and anomic aphasia, with varying times of stroke installation and GC consisting of 30 cognitively healthy participants. Computerized technologies were explored for the analysis of metrics related to the micro and macrostructure of discourses uttered from Cinderela history and Cookie Theft picture. RESULTS: Comparing the GC and GA2, in relation to the discourse macrostructure, it was observed that the GA2 aphasics differed significantly from the GC in relation to the total number of propositions emitted; considering the microstructure, seven metrics differentiated both groups. There was a significant difference in the macro and microstructure between the discourses of Broca\'s aphasic subjects and anomic ones. It was possible to verify differences in macro and microstructure measurements in GA1 with the advancement of injury time. In GA1, the comparison between parameters in the sub-acute phase and after 6 months of stroke revealed differences in macrostructure - increase in the number of propositions of the orientation block and of the total propositions. Regarding the microstructure, the initial measures of syllable metrics by word content, incidence of nouns and incidence of content words differed after 6 months of intervention. The variable incidence of missing words in the dictionary showed a significantly lower value after three months of stroke. Cinderella\'s story provided more complete microstructure data than the Cookie Theft picture. There was no change in SPECT over time, without demonstration of change with the evolution of aphasia. CONCLUSION: The discourse produced from the history of Cinderella and the Cookie Theft picture generated material for macrostructure and microstructure analysis of cognitively healthy and aphasic individuals, made it possible to quantify and qualify the evolution of language in different phases of stroke recuperation and distinguished the behavior of healthy and with Broca´s and anomic aphasia, in macro and microstructure aspects. The exploration of computerized tools facilitated the analysis of the data in relation to the microstructure, but it was not applicable to the macrostructure, demonstrating that there is a need for tool adjustments for the discourse analysis of patients. SPECT data did not reflect the behavioral improvement of the language of aphasic subjects

INTRODUÇÃO: O tema de investigação, discurso de afásicos, fornece informações importantes sobre aspectos fonológicos, morfológicos, sintáticos, semânticos e pragmáticos da linguagem de pacientes que sofreram lesão vascular cerebral. Uma das maneiras de estudar o discurso é por meio de cenas figurativas temáticas simples ou em sequência. A sequência da história de \"Cinderela\" é frequentemente utilizada em estudos, por ser familiar em todo o mundo, o que favorece estudos transculturais; por induzir a produção de narrativas, ao invés de descrições, frequentemente obtidas quando se utiliza prancha única para eliciar discursos. Outra vantagem do uso das sequências da \"Cinderela\" é o fato de gerar material linguístico em quantidade suficiente para análise detalhada. OBJETIVOS: (1) analisar, por meio de tecnologias computadorizadas, aspectos macro e microestruturais do discurso de indivíduos sadios do ponto de vista cognitivo, afásicos de Broca e afásicos anômicos; (2) explorar o discurso como indicador de evolução da afasia; (3) analisar a contribuição do SPECT para verificação de evolução da afasia junto ao discurso. MÉTODO: Participaram do estudo oito indivíduos afásicos de Broca e anômicos que compuseram o grupo do estudo longitudinal (G1), 15 indivíduos afásicos de Broca e anômicos que compuseram o outro grupo de estudo (G2) e 30 cognitivamente sadios (GC). Os participantes foram solicitados a examinar as cenas da história \"Cinderela\" e depois recontar a história, com suas palavras. Foram exploradas tecnologias computadorizadas e analisados aspectos macro e microestruturais dos discursos produzidos. Para o G1, tivermos a particularidade de coleta de discurso também pela prancha \"Roubo dos Biscoitos\", análise do exame SPECT e acompanhamento longitudinal por um período de seis meses. RESULTADOS: Comparando o GC e o G2, em relação à macroestrutura, notou-se que os afásicos do G2 se diferenciaram significativamente do GC em todas as proposições e, em relação à microestrutura, sete métricas foram capazes de diferenciar ambos os grupos. Houve diferença significante macro e micro estrutural entre os sujeitos afásicos de Broca e anômicos. Foi possível verificar diferenças em medidas da macro e da microestrutura no G1 com o avançar do tempo de lesão após AVC. A história da \"Cinderela\" forneceu dados de microestrutura mais completos do que a prancha \"Roubo dos Biscoitos\". Os resultados do SPECT permaneceram os mesmos, sem demonstração de mudança com a evolução da afasia. CONCLUSÃO: A produção de narrativa gerou material para análise de macroestrutura e microestrutura, tanto aspectos de macro quanto de microestrutura diferenciaram indivíduos cognitivamente sadios dos sujeitos afásicos. A análise do discurso da \"Cinderela\" serviu como instrumento para mensurar a melhora da linguagem dos sujeitos afásicos. O uso da ferramenta computacional auxiliou as análises discursivas / INTRODUCTION: The aphasic discourse analysis provides important information about the phonological, morphological, syntactic, semantic and pragmatic aspects of the language of patients who have suffered a stroke. The evaluation of the discourse, along with other methods, can contribute to observation of the evolution of the language and communication of aphasic patients; however, manual analysis is laborious and can lead to errors. OBJECTIVES: (1) to analyze, by computerized technologies, macro and microstructural aspects of the discourse of healthy cognitive individuals, Broca\'s and anomic aphasics; (2) to explore the discourse as indicator of the evolution of aphasia; (3) to analyze the contribution of single photon emission computed tomography (SPECT) to verify the correlation between behavioral and neuroimaging evolution data. METHOD: Two groups of patients were studied: GA1, consisting of eight individuals with Broca\'s aphasia and anomic aphasia, who were analyzed longitudinally from the sub-acute phase of the lesion and after three and six months; GA2 composed of 15 individuals with Broca\'s and anomic aphasia, with varying times of stroke installation and GC consisting of 30 cognitively healthy participants. Computerized technologies were explored for the analysis of metrics related to the micro and macrostructure of discourses uttered from Cinderela history and Cookie Theft picture. RESULTS: Comparing the GC and GA2, in relation to the discourse macrostructure, it was observed that the GA2 aphasics differed significantly from the GC in relation to the total number of propositions emitted; considering the microstructure, seven metrics differentiated both groups. There was a significant difference in the macro and microstructure between the discourses of Broca\'s aphasic subjects and anomic ones. It was possible to verify differences in macro and microstructure measurements in GA1 with the advancement of injury time. In GA1, the comparison between parameters in the sub-acute phase and after 6 months of stroke revealed differences in macrostructure - increase in the number of propositions of the orientation block and of the total propositions. Regarding the microstructure, the initial measures of syllable metrics by word content, incidence of nouns and incidence of content words differed after 6 months of intervention. The variable incidence of missing words in the dictionary showed a significantly lower value after three months of stroke. Cinderella\'s story provided more complete microstructure data than the Cookie Theft picture. There was no change in SPECT over time, without demonstration of change with the evolution of aphasia. CONCLUSION: The discourse produced from the history of Cinderella and the Cookie Theft picture generated material for macrostructure and microstructure analysis of cognitively healthy and aphasic individuals, made it possible to quantify and qualify the evolution of language in different phases of stroke recuperation and distinguished the behavior of healthy and with Broca´s and anomic aphasia, in macro and microstructure aspects. The exploration of computerized tools facilitated the analysis of the data in relation to the microstructure, but it was not applicable to the macrostructure, demonstrating that there is a need for tool adjustments for the discourse analysis of patients. SPECT data did not reflect the behavioral improvement of the language of aphasic subjects

INTRODUÇÃO: O envelhecimento da população é uma tendência social conhecida em países desenvolvidos e cada vez mais pronunciada em países em desenvolvimento. A demência é considerada um dos principais problemas de saúde devido ao rápido crescimento populacional de idosos, sendo os distúrbios de linguagem considerados importantes nesses quadros. O discurso tem ganhado destaque para a identificação dos distúrbios linguísticos nas demências assim como no seguimento desses pacientes. A caracterização das diferenças pode auxiliar no diagnóstico diferencial e contribuir para a criação de ferramentas futuras que auxiliem na intervenção clínica e ajudem a evitar a evolução e/ou progressão dos quadros demenciais. O processo de transcrição e análise do discurso é bastante laborioso, desta forma o uso de métodos computacionais tem auxiliado na identificação e extração de características linguísticas. OBJETIVO: identificar alterações em aspectos micro e macrolinguísticos que diferenciem indivíduos com doença de Alzheimer, comprometimento cognitivo leve e idosos sem comprometimento cognitivo na tarefa de narrativa de figuras em sequência e explorar a ferramenta computacional (Coh-Metrix-Dementia) para análise do discurso desses sujeitos. MÉTODO: Foram avaliados 60 indivíduos, sendo 20 em cada grupo de pesquisa (doença de Alzheimer leve - GDA, comprometimento cognitivo leve amnéstico - GCCLa e controle - GC). Os indivíduos foram solicitados a enunciar uma narrativa baseada em 22 cenas em sequência, que retratam a história da \"Cinderela\". Foram aplicados também os seguintes testes linguístico-cognitivos: Fluência Verbal, Teste de Nomeação do Boston e Camel and Cactus test. Utilizou-se o Coh-Metrix- Dementia para extração automática das métricas. RESULTADOS: Os valores extraídos pelo Coh-Metrix-Dementia foram tratados estatisticamente sendo possível levantar métricas capazes de distinguir os grupos estudados. Em relação aos aspectos microlinguísticos destacaram-se a redução nas habilidades sintáticas, maior dificuldade no resgate verbal, discursos com menor coesão e coerência local no GDA. No nível macrolinguístico o GDA apresentou os discursos menos informativos, com maior prejuízo em relação a coerência global e maior número de modalizações. O GDA também apresentou maior comprometimento da estrutura narrativa. Não foi possível discriminar o GCCLa e GC em nenhuma métrica do discurso deste estudo. Foram feitas adaptações em relação a segmentação das sentenças para um melhor funcionamento da ferramenta computacional. CONCLUSÃO: Os indivíduos do GDA apresentaram discursos com maior comprometimento macro e microestrutural. O uso da ferramenta computacional se mostrou um importante aliado para análises discursivas / INTRODUCTION: Population aging is a social trend known in developed countries and increasingly pronounced in developing countries. Dementia is considered one of the main health problems due to the rapid population growth of the elderly, and language disorders are considered important in these settings. The discourse is important for the identification of linguistic disorders in dementias as well as in the follow-up of these patients. The discourse differences characterization can help on the differential diagnosis and contribute to the creation of future tools for clinical intervention and help prevent the evolution and/or progression of dementia. The transcription and discourse analysis are laborius, thus the use of computational methods helped in the identification and extraction of linguistic characteristics. OBJECTIVE: The objective of this study was to identify changes in micro and macrolinguistic aspects that differentiate individuals with Alzheimer\'s disease, mild cognitive impairment and healthy elderly individuals during narrative of figures in sequence and to explore the computational tool (Coh-Metrix-Dementia) to analyze the subjects\' discourse. METHODS: 60 subjects were evaluated, 20 of them in each research group (mild Alzheimer\'s disease - GDA, amnestic cognitive impairment - GCCLa and control - CG). The subjects were asked to construct a narrative based on sequence of pictures, about the \"Cinderella´s Story\". The following linguistic-cognitive tests were also applied: Verbal Fluency, Boston Naming Test, and Camel and Cactus test. Coh-Metrix-Dementia was used for automatic metrics extraction. RESULTS: The values extracted by Coh-Metrix-Dementia were statistically treated and it was possible to obtain metrics capable of distinguishing the studied groups. In relation to the microlinguistic aspects, it was found the reduction in syntactic abilities, greater difficulty in verbal rescue, discourses with less cohesion and local coherence in the GDA. In the macrolinguistic level the GDA presented the less informative discourses, with greater loss in global coherence and the greater number of modalizations. The GDA also presented greater impairment on narrative structure. It was not possible to discriminate GCCLa and GC in any discourse´s metric in this study tool functioning. CONCLUSION: The GDA subjects presented discourses with greater macro and microstructural impairment. The computational tool usage proved to be an important ally for discursive analysis

Title: Automated determination of earthquake source parameters Author: Jiří Vackář Department: Department of Geophysics Supervisor: prof. RNDr. Jiří Zahradník, DrSc., Department of Geophysics Abstract: The thesis deals with methods for automated inversion of seismic source parameters. We studied the influence of structure model used and show an ex- ample how the existing model can be improved. We have developed a new, fully automated tool for the centroid moment tensor (CMT) inversion in a Bayesian framework. It includes automated data retrieval from ArcLink server or local data storage. Step-like disturbances are detected using modeling of the distur- bance according to instrument parameters and such components are automati- cally excluded from further processing. Frequency ranges for the filtration and time windows for the inversion are determined automatically due to epicentral distance. Full-waveform inversion is performed in a space-time grid around a provided hypocenter. A data covariance matrix calculated from pre-event noise yields an automated weighting of the station recordings according to their noise levels and also serves as an automated frequency filter suppressing noisy frequency ranges. The method is tested on synthetic and observed data. It is applied on a dataset from the Swiss seismic...