Orientador: Ricardo Pasquini / Co-orientador: Luiz Fernando Bleggi Torres / Dissertação(mestrado) - Universidade Federal do Parana, Setor de Ciencias da Saude, Programa de Pós-Graduação em Medicina Interna

INTRODUÇÃO: O emprego de novas tecnologias na reabilitação de linguagem pode auxiliar amplamente o processo terapêutico mediante treino intensivo. Para isso, no entanto, é necessário que essas ferramentas atendam às demandas desses pacientes, e que o terapeuta seja protagonista no desenvolvimento dessas tecnologias. Essas ferramentas precisam ainda ser analisadas por meio de pesquisas científicas que avaliem sua efetividade no tratamento fonoaudiológico. OBJETIVOS: Desenvolver uma plataforma on-line para reabilitação de pacientes afásicos e avaliar sua eficácia em dois experimentos. MÉTODOS: Essa pesquisa foi dividida em três estudos: Estudo 1) criação da plataforma com emprego do modelo de desenvolvimento ágil e com uso de software livre, e aplicação das normas de qualidade ISO 25010. Estudo 2) estudo prospectivo, longitudinal e intervencionista, com a finalidade de prover treino de aprendizado de palavras, para avaliar as opiniões de 12 pessoas idosas cognitivamente sadias sobre o uso da plataforma. Os participantes foram divididos em três grupos: grupo controle (GC); grupo experimento escrita (GEE); grupo experimento repetição (GER). O treino foi conduzido na plataforma e foi composto por atividades de treino com emprego de 60 pseudopalavras, associadas a figuras. O estudo teve duração de um mês para cada participante, sendo composto por três etapas: 1) Pré-treino (1 dia); 2) Treino (5 dias) e 3) Pós-treino (3 dias). As etapas 1 e 3 foram comuns para todos os participantes. Na etapa 1, os participantes realizaram duas atividades a plataforma com o propósito de apresentar a plataforma e os estímulos usados nas outras etapas do estudo. Na etapa 2, os GEE e GER realizaram o treino de pseudopalavras, enquanto o GC realizou atividade de jogos. A etapa 3 foi dividida em três avaliações: após os cinco dias de treino; uma semana após a primeira avaliação e 1 mês depois do início do treino. A análise estatística avaliou a acurácia (número de acertos); efeitos de idade, escolaridade, intensidade e modalidade do treino; e o impacto de propriedades dos estímulos. O Estudo 3 apresenta o relato de caso de uma paciente com afasia de Wernicke grave, que usou a plataforma intensamente ao longo de 18 meses de terapia de linguagem. RESULTADOS: Estudo 1) Criação da plataforma com elaboração de 114 atividades terapêuticas, com exportação de dados para análise estatística e adequação às normas de qualidade ISO 25010. Estudo 2) Os participantes dos grupos GEE e GER apresentaram desempenho significativamente superior ao do GC em todas as etapas do pós-treino. Todos os participantes apresentaram opiniões favoráveis sobre a plataforma. Estudo 3) A paciente apresentou melhora significativa em habilidades linguísticas, com uso de 58 atividades da plataforma, totalizando 280 horas de treino com a ferramenta. CONCLUSÕES: A plataforma mostrou-se eficiente para atender às necessidades de pessoas idosas cognitivamente sadias e na reabilitação de linguagem / INTRODUCTION: The application of new technologies in language rehabilitation has the potential to improve therapies through intensive training. But, to that end, it is important that these tools meet the needs of patients and that the therapist becomes a main player during the development of these technologies. These tools also need to be evaluated by means of scientific research that evaluate their effectiveness on the speech and language pathology treatment. OBJECTIVE: To develop an online platform for the rehabilitation of aphasic patients, and to evaluate its effectiveness in two experiments. METHODS: This research was divided into three studies: Study 1) creation of the platform using the agile development model and free software, and application of ISO 25010 quality standards. Study 2) a prospective, longitudinal and interventional study with the purpose of providing training in word learning, in order to evaluate the opinions of 12 cognitively healthy elderly people on the use of the platform. Participants were divided into three groups: control group (CG), writing experiment group (WEG), and repetition experiment group (REG). The training was conducted on the platform and was composed of training activities with the employment of 60 pseudowords, associated with pictures. The study had a duration of one month for each participant, and was composed of three stages: 1) Pre-training (1 day), 2) Training (5 days), and 3) Post-training (3 days). Stages 1 and 3 were common to all participants. In the stage 1, the participants performed two activities in platform in order to present the platform and the stimulus used in the other stages. In stage 2, the WEG and REG performed pseudoword training; and the CG performed gaming activity. Stage 3 was divided into three evaluations: a) after five days of training, b) one week after the first evaluation, and one month after the beginning of training. The statistical analysis evaluated the accuracy (number of hits); effects of age, schooling, intensity, and training modality; and the impact of stimulus properties. Study 3) presented the case report of a patient with severe Wernicke\'s aphasia who used intensively the platform over 18 months of language therapy. RESULTS: Study 1) Creation of the platform with elaboration of 114 therapeutic activities, with export of data for statistical analysis and adaptation to ISO 25010 quality standards. Study 2) Participants in the WEG and REG groups presented performance significantly superior to the CG in all post-training steps. All participants presented favorable opinions on the platform. Study 3) The patient presented significant improvement in language skills with the use of 58 platform activities, totaling 280 hours of training with the tool. CONCLUSIONS: The platform proved to be an efficient tool in meeting the needs of cognitively healthy older people and in language rehabilitation

INTRODUÇÃO: O emprego de novas tecnologias na reabilitação de linguagem pode auxiliar amplamente o processo terapêutico mediante treino intensivo. Para isso, no entanto, é necessário que essas ferramentas atendam às demandas desses pacientes, e que o terapeuta seja protagonista no desenvolvimento dessas tecnologias. Essas ferramentas precisam ainda ser analisadas por meio de pesquisas científicas que avaliem sua efetividade no tratamento fonoaudiológico. OBJETIVOS: Desenvolver uma plataforma on-line para reabilitação de pacientes afásicos e avaliar sua eficácia em dois experimentos. MÉTODOS: Essa pesquisa foi dividida em três estudos: Estudo 1) criação da plataforma com emprego do modelo de desenvolvimento ágil e com uso de software livre, e aplicação das normas de qualidade ISO 25010. Estudo 2) estudo prospectivo, longitudinal e intervencionista, com a finalidade de prover treino de aprendizado de palavras, para avaliar as opiniões de 12 pessoas idosas cognitivamente sadias sobre o uso da plataforma. Os participantes foram divididos em três grupos: grupo controle (GC); grupo experimento escrita (GEE); grupo experimento repetição (GER). O treino foi conduzido na plataforma e foi composto por atividades de treino com emprego de 60 pseudopalavras, associadas a figuras. O estudo teve duração de um mês para cada participante, sendo composto por três etapas: 1) Pré-treino (1 dia); 2) Treino (5 dias) e 3) Pós-treino (3 dias). As etapas 1 e 3 foram comuns para todos os participantes. Na etapa 1, os participantes realizaram duas atividades a plataforma com o propósito de apresentar a plataforma e os estímulos usados nas outras etapas do estudo. Na etapa 2, os GEE e GER realizaram o treino de pseudopalavras, enquanto o GC realizou atividade de jogos. A etapa 3 foi dividida em três avaliações: após os cinco dias de treino; uma semana após a primeira avaliação e 1 mês depois do início do treino. A análise estatística avaliou a acurácia (número de acertos); efeitos de idade, escolaridade, intensidade e modalidade do treino; e o impacto de propriedades dos estímulos. O Estudo 3 apresenta o relato de caso de uma paciente com afasia de Wernicke grave, que usou a plataforma intensamente ao longo de 18 meses de terapia de linguagem. RESULTADOS: Estudo 1) Criação da plataforma com elaboração de 114 atividades terapêuticas, com exportação de dados para análise estatística e adequação às normas de qualidade ISO 25010. Estudo 2) Os participantes dos grupos GEE e GER apresentaram desempenho significativamente superior ao do GC em todas as etapas do pós-treino. Todos os participantes apresentaram opiniões favoráveis sobre a plataforma. Estudo 3) A paciente apresentou melhora significativa em habilidades linguísticas, com uso de 58 atividades da plataforma, totalizando 280 horas de treino com a ferramenta. CONCLUSÕES: A plataforma mostrou-se eficiente para atender às necessidades de pessoas idosas cognitivamente sadias e na reabilitação de linguagem / INTRODUCTION: The application of new technologies in language rehabilitation has the potential to improve therapies through intensive training. But, to that end, it is important that these tools meet the needs of patients and that the therapist becomes a main player during the development of these technologies. These tools also need to be evaluated by means of scientific research that evaluate their effectiveness on the speech and language pathology treatment. OBJECTIVE: To develop an online platform for the rehabilitation of aphasic patients, and to evaluate its effectiveness in two experiments. METHODS: This research was divided into three studies: Study 1) creation of the platform using the agile development model and free software, and application of ISO 25010 quality standards. Study 2) a prospective, longitudinal and interventional study with the purpose of providing training in word learning, in order to evaluate the opinions of 12 cognitively healthy elderly people on the use of the platform. Participants were divided into three groups: control group (CG), writing experiment group (WEG), and repetition experiment group (REG). The training was conducted on the platform and was composed of training activities with the employment of 60 pseudowords, associated with pictures. The study had a duration of one month for each participant, and was composed of three stages: 1) Pre-training (1 day), 2) Training (5 days), and 3) Post-training (3 days). Stages 1 and 3 were common to all participants. In the stage 1, the participants performed two activities in platform in order to present the platform and the stimulus used in the other stages. In stage 2, the WEG and REG performed pseudoword training; and the CG performed gaming activity. Stage 3 was divided into three evaluations: a) after five days of training, b) one week after the first evaluation, and one month after the beginning of training. The statistical analysis evaluated the accuracy (number of hits); effects of age, schooling, intensity, and training modality; and the impact of stimulus properties. Study 3) presented the case report of a patient with severe Wernicke\'s aphasia who used intensively the platform over 18 months of language therapy. RESULTS: Study 1) Creation of the platform with elaboration of 114 therapeutic activities, with export of data for statistical analysis and adaptation to ISO 25010 quality standards. Study 2) Participants in the WEG and REG groups presented performance significantly superior to the CG in all post-training steps. All participants presented favorable opinions on the platform. Study 3) The patient presented significant improvement in language skills with the use of 58 platform activities, totaling 280 hours of training with the tool. CONCLUSIONS: The platform proved to be an efficient tool in meeting the needs of cognitively healthy older people and in language rehabilitation

In dieser Arbeit wurde die systematische Suche nach krankheitsassoziierten Genen bei periodischer Katatonie fortgeführt. Für diese Erkrankung war die klinische Abgrenzbarkeit und die familiäre Häufung signifikant und ließ aufgrund der vertikalen Transmission und dem Auftreten über mehrere Generationen und hinweg auf einen Hauptgeneffekt schließen. Nach der Durchführung von Kopplungs-Analysen kristallisierten sich zwei koppelnde Regionen auf den Chromosomen 15 und 22 heraus. Mittels Haplotypanalyse konnte der Genort auf Chromosom 22q13 auf einen knapp 5 Mbp großen Bereich eingeschränkt werden. Im kodierenden Bereich des MLC1-Genes segregierte im mit periodischer Katatonie assoziierten Haplotyp eine Variante (p.Leu309Met). Da Mutationen im MLC1-Gen bereits im Zusammenhang mit Megalenzephaler Leukoenzephalopathie beschrieben worden waren, wurden in dieser Arbeit zunächst fünf Patienten mit dieser Erkrankung auf Mutationen in kodierenden Bereichen von MLC1 systematisch untersucht. Daran schloss sich eine Analyse dieses Gens bei 140 Patienten mit periodischer Katatonie an. Ein Zusammenhang zwischen Mutationen in MLC1 und dem Auftreten von Megalenzephaler Leukoenzephalopathie wurde untermauert, wohingegen die Ergebnisse eindeutig gegen eine Assoziation mit periodischer Katatonie sprachen. Ein weiteres im Gehirn exprimiertes Kandidatengen (KIAA0767/DIP) wurde in dieser Arbeit untersucht. Dabei wurden sechs SNPs im exonnahen intronischen Bereich entdeckt sowie eine Variante im Exonbereich (p.Glu156Asn). Dies ist eine seltene Normvariante, eine Assoziation zur periodischen Katatonie wurde in einer Fall-Kontroll-Studie ausgeschlossen. Insgesamt wurde durch die systematische Mutationsanalyse die Kandidatenregion auf Chromosom 22q13.3 weiter eingeengt. Gegen einen Zusammenhang zwischen MLC1 und periodischer Katatonie sprechen die vorgestellten validen Ergebnisse. / The present dissertation has carried on the systematic screening for genes involved in periodic catatonia. The clinical definition and the familial clustering of this disorder were of statistical significance. Its vertical transmission and the occurrence across generations led to the conclusion of it being a major gene effect. Linkage scans discovered two linking regions on chromosome 15 and chromosome 22. Using haplotype analysis it was possible to restrict the candidate region on chromosome 22q13 to region of 5 Mbp. In the coding region of the MLC1 gene a variant (p.Leu309Met) segregated in the haplotype associated with periodic catatonia. As detailed descriptions of mutations in the MLC1 gene in connection with megalencephalic leucoencephalopathy already exist, for the present dissertation five patients with the aforesaid disorder were systematically screened for mutations in the coding regions of MLC1. This was followed by an analysis of the according gene in 140 patients with periodic catatonia, which substantiated a correlation between mutations in MLC1 and the occurrence of megalencephalic leucoencephalopathy, whereas the results definitely did not support an association with periodic catatonia. Additionally, a further brain-expressed candidate gene (KIAA0027/DIP) was screened in this dissertation discovering six single nucleotide polymorphisms in the intronic exon-near region and a variant in the exonic region (p.Glu156Asn). This is a rare normal variant, an association with periodic catatonia was excluded in a case-control study. Altogether the systematic mutation analysis enabled a further reduction of the candidate region on chromosome 22q13.3. The research results, which meet the criterion of validity, do not support a correlation between MLC1 and periodic catatonia.

Doctor of Philosophy (PhD) / Proteomics is rapidly achieving recognition as a complimentary and perhaps superior approach to examine global changes in protein abundance in complex biological systems and the value of these techniques in neuropsychiatry is beginning to be acknowledged. Characterizing the brain’s regional proteomes provides a foundation for the detection of proteins that may be involved in disease-related processes. Firstly, optimal conditions were achieved for the application of two dimensional-gel electrophoresis (2D-GE)-based proteomics with postmortem human brain tissue. These optimized techniques were then applied to soluble fractions of adjacent grey and white matter of a single cytoarchitecturally defined area (Brodmann area 9; BA9) and of two adjacent regions of frontal white matter (BA9 and CC body) from healthy individuals. These normative proteomic comparisons highlighted the importance of correct tissue sampling, i.e. proper separation of regional white matter, as heterogeneity in the respective proteomes was demonstrated. Furthermore, they stressed the necessity for future molecular brain mapping studies. The main focus of this thesis however, was to examine the proteomes of brain regions specifically vulnerable to alcohol-induced damage underlying cognitive dysfunction. Alcoholic patients commonly experience mild to severe cognitive decline. It is postulated that cognitive dysfunction is caused by an alcohol-induced region selective brain damage, particularly to the prefrontal cortex. The cerebellum is increasingly recognized for its role in various aspects of cognition and alcohol–induced damage to the cerebellar vermis could indirectly affect neurocognitive functions attributed to the frontal lobe. We used a 2D-GE-based proteomics approach to compare protein abundance profiles of BA9 grey and white matter and the cerebellar vermis from human alcoholics (neurologically uncomplicated and alcoholics complicated with liver cirrhosis) and healthy control brains. Among the protein level changes observed are disturbances in the levels of a number of thiamine-dependent enzymes. A derangement in energy metabolism perhaps related to thiamine deficiency seems to be important in all regions analysed, even where there are no clinical or pathological findings of Wernicke-Korsakoff Syndrome. Evidence of oxidative changes was also seen in all regions and effects of liver dysfunction in the vermis found. However, overall, these results highlight the complexity of this disease process in that a number of different proteins from different cellular pathways appear to be affected. By identifying changes in protein abundance levels in the prefrontal grey and white matter and the cerebellar vermis, hypotheses may draw upon more mechanistic explanations as to how chronic ethanol consumption causes the structural and functional alterations associated with alcohol-related brain damage. Furthermore, by comparing these results, we may be able to isolate disturbances in molecular pathways specific to the brain damage caused by alcohol, severe liver dysfunction and thiamine deficiency.

No description available.

Wernicke's encephalopathy is a neuropsychiatric disorder resulting from thiamine deficiency (TD) and is characterized by neuronal loss, astrocytic proliferation and microglial activation. Cyclooxygenases (COX) are enzymes which catalyze the first step in the synthesis of prostanoids. COX-1 is expressed constitutively and COX-2 is the inducible isoform. Groups of TD rats and pair-fed controls were killed at presymptomatic and symptomatic stages of encephalopathy. Cresyl violet and NeuN staining showed decreased numbers of neuronal cells in vulnerable regions (medial thalamus and inferior colliculus) but not in a spared region (frontal cortex). Numbers of GFAP-positive and OX-42-positive cells were increased at symptomatic stage of encephalopathy. Expression of COX-2 mRNA and neuronal COX-2 immunoreactivity were selectively increased in vulnerable regions of TD rats at symptomatic stages of encephalopathy. Nimesulide, a highly selective COX-2 inhibitor, lowered PGE2 levels and precipitated the progression of encephalopathy suggesting that COX-2 in this model is conferring neuroprotection.

Doctor of Philosophy (PhD) / Proteomics is rapidly achieving recognition as a complimentary and perhaps superior approach to examine global changes in protein abundance in complex biological systems and the value of these techniques in neuropsychiatry is beginning to be acknowledged. Characterizing the brain’s regional proteomes provides a foundation for the detection of proteins that may be involved in disease-related processes. Firstly, optimal conditions were achieved for the application of two dimensional-gel electrophoresis (2D-GE)-based proteomics with postmortem human brain tissue. These optimized techniques were then applied to soluble fractions of adjacent grey and white matter of a single cytoarchitecturally defined area (Brodmann area 9; BA9) and of two adjacent regions of frontal white matter (BA9 and CC body) from healthy individuals. These normative proteomic comparisons highlighted the importance of correct tissue sampling, i.e. proper separation of regional white matter, as heterogeneity in the respective proteomes was demonstrated. Furthermore, they stressed the necessity for future molecular brain mapping studies. The main focus of this thesis however, was to examine the proteomes of brain regions specifically vulnerable to alcohol-induced damage underlying cognitive dysfunction. Alcoholic patients commonly experience mild to severe cognitive decline. It is postulated that cognitive dysfunction is caused by an alcohol-induced region selective brain damage, particularly to the prefrontal cortex. The cerebellum is increasingly recognized for its role in various aspects of cognition and alcohol–induced damage to the cerebellar vermis could indirectly affect neurocognitive functions attributed to the frontal lobe. We used a 2D-GE-based proteomics approach to compare protein abundance profiles of BA9 grey and white matter and the cerebellar vermis from human alcoholics (neurologically uncomplicated and alcoholics complicated with liver cirrhosis) and healthy control brains. Among the protein level changes observed are disturbances in the levels of a number of thiamine-dependent enzymes. A derangement in energy metabolism perhaps related to thiamine deficiency seems to be important in all regions analysed, even where there are no clinical or pathological findings of Wernicke-Korsakoff Syndrome. Evidence of oxidative changes was also seen in all regions and effects of liver dysfunction in the vermis found. However, overall, these results highlight the complexity of this disease process in that a number of different proteins from different cellular pathways appear to be affected. By identifying changes in protein abundance levels in the prefrontal grey and white matter and the cerebellar vermis, hypotheses may draw upon more mechanistic explanations as to how chronic ethanol consumption causes the structural and functional alterations associated with alcohol-related brain damage. Furthermore, by comparing these results, we may be able to isolate disturbances in molecular pathways specific to the brain damage caused by alcohol, severe liver dysfunction and thiamine deficiency.

Thesis (Ph. D.)--State University of New York at Binghamton, Department of Psychology, Behavioral Neuroscience, 2008. / Includes bibliographical references.

Thesis (M.A.)--State University of New York at Binghamton, Department of Psychology, 2005. / Includes bibliographical references.