INTRODUÇÃO: Epilepsia do lobo temporal (ELTM) é a forma mais comum de epilepsia parcial, ocorrendo em cerca de 40% dos pacientes. Na ELTM as crises epilépticas podem ter origem na região cortical, o que é menos comum, porém ocorrem frequentemente em estruturas mesiais do lobo temporal, representadas basicamente pelo hipocampo e/ou amígdala. Com intuito de incluir novas possibilidades de tratamento e diagnóstico, torna-se necessário uma maior compreensão das bases moleculares da ELMT. Dentro desta perspectiva, um dos grandes desafios para a Neurociências na atualidade é o desenvolvimento de biomarcadores que facilitem o diagnóstico e prognóstico para a epilepsia. Recentemente, com o desenvolvimento de algumas pesquisas e novas técnicas no campo da biologia molecular demonstram que microRNAs circulantes no sangue são biomarcadores sensíveis e específicos para várias doenças, incluindo doenças neurológicas, podendo ser obtido de forma não invasiva, representando assim um método de eficiente detecção e custo baixo. OBJETIVOS: Analisar o perfil de expressão dos microRNAs miR-629-3p, miR-1202, miR1225-5p na amígdala, hipocampo e sangue de pacientes submetidos a cirurgia para tratamento da epilepsia do lobo temporal mesial refratários ao tratamento medicamentoso e investigar se os mesmos podem auxiliar como biomarcadores de diagnóstico e prognóstico para a epilepsia. PACIENTES E MÉTODOS: Foram utilizadas amostras de amígdalas, hipocampos e sangue de 20 pacientes com ELTM, sendo 10 com boa evolução pós-operatória (Engel I) e 10 com evolução pós-operatória insatisfatória (Engel III e IV), e para controle foram utilizados 10 amostras de amígdalas e 10 de hipocampos de necrópsias, assim como 10 amostras de sangue de indivíduos saudáveis. A análise de expressão dos miRNAs foi feita utilizando a técnica de RQPCR. RESULTADOS e CONCLUSÕES: Os miRNAs miR-629-3p, miR-1202 e miR- 1225-5p apresentaram-se hiperexpressos no sangue de pacientes com ELTM, podendo sugerir um possível papel de biomarcadores auxiliando no diagnóstico da ELTM. Os miRNAs-629-3p, 1202 e 1225-5p apresentaram aumento nos níveis de expressão sanguíneos progressivos entre os grupos controle, Engel I e Engel III e IV respectivamente, apresentando também possível potencial de biomarcadores no prognóstico cirúrgico entre Engel I e Engel III e IV. / INTRODUCTION: Epilepsy of the temporal lobe is the most common form of partial epilepsy, occurring in about 40% of patients. In TLE epileptic seizures may originate in the cortical region, which is less common. However they occur more frequently in temporal lobe mesial structures, represented primarily by the hippocampus and / or amygdala. A greater understanding of the epilepsy molecular basis is necessary when aiming to implement new treatments possibilities. Within this field of study, one of the major challenges for neuroscience today is the development of biomarkers that facilitate the diagnosis and prognosis for epilepsy. Recently, with the development of some researches and new techniques in the field of molecular biology, it demonstrate that microRNAs circulating in the blood are sensitive and specific biomarkers for various diseases, including neurological diseases and they can be obtained noninvasively, thus representing a low cost method of efficient detection. OBJECTIVES: To analyze the expression profile of miR-629-3p, miR-1202, miR1225-5p microRNAs in the amygdala, hippocampus and blood of patients operated to treat the mesial temporal lobe epilepsy refractory to drug treatment and investigate whether they can be used as biomarkers for epilepsy diagnosis and prognosis. PATIENTS AND METHODS: Amygdala, hippocampus and blood samples of 20 patients with MTLE were used; 10 with good postoperative outcome (Engel I) and 10 with poor postoperative outcome (Engel III and IV), and for the control group 10 amygdalas and 10 hippocampus of necropsy, as well as, 10 samples blood from healthy individuals. The analysisof the expression of miRNAs was performed using RQ-PCR. RESULTS AND CONCLUSION: MiRNAs miR-629-3P, miR-1202 and miR- 1225-5P were hyper-expressed in the blood of patients with MTLE, and may suggest a possible role of biomarkers in the diagnosis of MTLE. miR-629-3p, miR-1202 and miR-1225-5p were progressively expressed in the blood of patients in the control, Engel I and Engel III and IV groups respectively, representing also potential biomarkers for surgical prognosis between Engel I and Engel III and IV.

A memória prospectiva (MP), função não avaliada em baterias neuropsicológicas tradicionalmente aplicadas em pacientes com epilepsia do lobo temporal secundária à esclerose de hipocampo (EMT), refere-se a um conjunto de habilidades cognitivas que permitem lembrar-se de uma intenção a desempenhar no futuro, no momento adequado. Estudos de neuroimagem associam os lobos frontais ao desempenho do componente prospectivo da MP (lembrar a intenção) e as estruturas mesiais temporais ao componente retrospectivo da MP (lembrar o conteúdo da atividade). Estudou-se a MP, com ênfase no componente prospectivo, em pacientes com EMT unilateral. Correlacionou-se o desempenho no teste de MP com o desempenho em testes neuropsicológicos tradicionalmente utilizados e escalas de auto-avaliação. Incluímos 26 pacientes com EMT à direita, 22 à esquerda e 26 controles sadios pareados por idade, sexo e escolaridade. Houve alteração do componente prospectivo da MP em ambos os grupos EMT (p<0,001) (efeito lesão), com pior resultado no grupo EMTE (p<0,05) (efeito lateralidade da lesão). O grupo EMTE também apresentou rebaixamento no componente retrospectivo da MP. A alteração do desempenho não pôde ser explicada por depressão, ansiedade, efeito das drogas antiepilépticas, número de crises ou duração da epilepsia. O desempenho de pacientes com EMTE no teste de MP correlacionou-se com o desempenho em testes de evocação tardia de material verbal e autopercepção de desempenho de memória, o que sugere substrato neurobiológico comum para as funções avaliadas por estes instrumentos. A bateria de MP demonstrou o efeito da lesão e da lateralidade da lesão para os erros de componente prospectivo, e o efeito da lateralidade da lesão para os erros de componente retrospectivo. Sugere-se que o papel de estruturas mesiais temporais, incluindo os hipocampos, sobre o componente prospectivo da MP deve ser considerado, especialmente em atividades de longo prazo. A inclusão da avaliação da MP em baterias neuropsicológicas para pacientes com EMT pode auxiliar o entendimento das dificuldades de memória que estes apresentam em seu cotidiano. / Prospective memory (PM) refers to a set of cognitive abilities that allow future performance of a present intention, at the appropriate time. It has not been evaluated in mesial temporal sclerosis (MTS). Functional neuroimaging studies indicate the role of the frontal lobes in the prospective component of PM (intention recall) and the role of mesial temporal structures in the retrospective component (content recall). We studied PM, with emphasis on the prospective component, in patients with unilateral MTS and correlated performance in PM testing with performance in traditional neuropsychological tests, as well as self evaluation scales. We evaluated PM in 26 right MTS and in 22 left MTS patients, as well as in 26 age-gender and education matched normal controls. The prospective component of PM was impaired in both patient groups (p<0.001) (lesion effect), worse in LMTS group (p< 0.05) (laterality effect). Left MTS also impacted on performance in the retrospective PM component. Impaired performance could not be explained by depression, anxiety, age at epilepsy onset, duration of epilepsy or an antiepileptic drug effect. Mesial temporal lobe structures play an important role in prospective PM processes, especially in tasks involving long delay intervals. Inclusion of PM tests in neuropsychological batteries used to evaluate patients with temporal lobe structures dysfunction may allow a better understanding of memory impairment in these patients, especially of the impact of PM impairment on daily living activities.

Introdução: Dois grandes estudos de genoma humano ENIGMA e CHARGE identificaram uma variante genética comum da tescalcina (TESCrs7294919) envolvida no volume hipocampal em indivíduos saudáveis. Sabe-se que o gene TESC é altamente expresso na embriogênese e no hipocampo adulto e pode ter papel chave em doenças que afetam o hipocampo, entre estas, a epilepsia do lobo temporal. Neste estudo será investigado o impacto do polimorfismo rs7294919 em pacientes com epilepsia do lobo temporal. Métodos: Estudo de associação de 162 pacientes com epilepsia do lobo temporal e 100 controles sem epilepsia. Foram comparadas as frequências das variantes rs7294919 entre pacientes com epilepsia e controles e avaliados os impactos das diferenças alélicas nas principais características de epilepsia. Resultados: Não houve diferenças significativas entre os polimorfismos rs7294919 entre pacientes e controles (Tabela 1). No entanto, o genótipo CT foi mais comum no sexo masculino do que do sexo feminino (Tabela 2, Tabela 3 e Figura 1). Não foram observadas outras diferenças em relação à média de idade de início da epilepsia, história familiar de epilepsia, presença de aura, controle das crises, anormalidades de EEG interictal e achados de neuroimagem. Depois de controle de variáveis confusionais, o alelo C esteve significativamente aumentado em pacientes do sexo masculino(OR = 2,57; IC = 1.26-5.24 ; p = 0,009). Conclusão:Aobservação de uma maior frequência do alelo do alelo C em homens pode sugerir um efeito dependente da tescalcina através do seu gene TESCsobre o desenvolvimento da epilepsia do lobo temporal. Tanto quanto sabemos, este é o primeiro estudo da tescalcina no desenvolvimento de epilepsia. / Rationale: Two independent large genome-wide association studies identified a common genetic variant of tescalcin polymorphism (TESC rs7294919), that is significantly associated with hippocampal volume in healthy subjects. It is known that the TESC gene is highly expressed in embryogenesis and in adult hippocampus and might play a role in characteristics of diseases that affect hippocampus, among them temporal lobe epilepsy. In this study we investigate the impact of rs7294919 variants in temporal lobe epilepsy. Methods: This is an association study of 162 patients with temporal lobe epilepsy and 100 controls without epilepsy. We compared frequencies of rs7294919 variant between patients with epilepsy and controls and evaluate the impact of allelic difference in main characteristics of epilepsy. Results: There were no significant differences between TESC gene polymorphisms of patients and controls (Table 1). However, genotype CT was more frequent in male than females (Table 2, Table 3 and Figure 1). No other differences were observed regarding mean age of epilepsy onset, family history of epilepsy, presence of aura, seizure control, EEG interictal abnormalities and neuroimaging findings. After controlling for confounding variables, C allele was increased in male patients (O.R. = 2.57; CI = 1.26-5.24; p=0.009. Conclusions: Our finding might suggest a gender dependent effect of tescalcin polymorphism (TESC rs7294919) on development of epilepsy. As far as we are aware, this is the first study of tescalcin polymorphism on development of epilepsy. In our view, other studies in this venue are necessary to confirm our findings and explore other possible effects of tescalcina polymorphisms on development and characteristics of epilepsy.

Introdução: A epilepsia do lobo temporal mesial (ELTM) é a forma mais comum de epilepsia parcial e com índices significativos de resistência aos tratamentos farmacológicos. Um dos grandes desafios para a neurociência na atualidade é o desenvolvimento de biomarcadores que facilitem o diagnóstico e prognóstico para a epilepsia. Recentemente, algumas pesquisas demonstraram que microRNAs circulantes no sangue são biomarcadores sensíveis e específicos para várias doenças, incluindo do SNC, podendo ser obtido de forma não invasiva e representando um método de detecção eficiente e de baixo custo. Portanto, nosso trabalho teve como objetivo analisar o perfil de expressão dos microRNAs miR-145, miR-181c, miR199a e miR-1183 no hipocampo e fração leucocitária do sangue de pacientes com ELTM e investigar se os mesmos podem auxiliar como biomarcadores de diagnóstico e prognóstico para a epilepsia. Pacientes e métodos: Foram utilizadas amostras de hipocampo e sangue de 20 pacientes com ELTM, sendo 10 com boa evolução pós-operatória (Engel I) e 10 com evolução pósoperatória insatisfatória (Engel III e IV), e para controle foram utilizados hipocampos de necropsias e sangue de indivíduos saudáveis. A análise de expressão dos miRNAs foi feita utilizando a técnica de RQ-PCR. Resultados e Conclusões: As expressões dos miRNAs se comportaram de forma diferente no hipocampo e sangue de pacientes com ELTM quando comparados aos grupos controles. O miR- 145 apresentou-se hipoexpresso no hipocampo e hiperexpresso no sangue. Os miRNAs miR-145, miR-181c, miR199a e miR-1183 apresentaram-se hiperexpressos na fração leucocitária do sangue de pacientes com ELTM, podendo estes contribuir como possíveis biomarcadores para auxiliarem no diagnóstico. Ainda, não encontramos diferenças estatística desses miRNAs no sangue e hipocampo de pacientes Engel I versus Engel III e IV, no entanto, o miR-1183 apresentou-se progressivamente mais expresso no sangue dos pacientes nos grupos controle, no grupo Engel I e no grupo Engel III e IV respectivamente. / Introduction: The mesial temporal lobe epilepsy (MTLE) is the most common form of partial epilepsy and also presents significant resistance to pharmacological treatment. A major challenge for neuroscience today is the development of biomarkers that facilitate the diagnosis and prognosis for epilepsy. Recently, research has shown that microRNAs circulating in the blood are sensitive and specific biomarkers for various diseases, including CNS, they can be obtained noninvasively and represent a low cost detection method. Thus, our study aimed to analyze the expression profile of microRNAs miR-145, miR-181c, miR199a and miR- 1183 in the hippocampus and leukocyte fraction of the blood of patients with MTLE and investigate whether they can assist as diagnostic and prognosis biomarkers for epilepsy. Patients and Methods: Hippocampus and blood samples of 20 patients with MTLE were used; ten with good postoperative outcome (Engel I) and ten with poor postoperative outcome (Engel III and IV), and for the control group hippocampus of necropsy and blood of healthy individuals were used. The analysis of the expression of miRNAs was performed using RQ-PCR. Results and Conclusions: The expression of miRNAs behaved differently in the hippocampus and blood of patients with MTLE when compared with the control groups. This different behavior was most evident in miR-145, which was hypo-expressed in the hippocampus and hyper-expressed in the blood. MiRNAs miR-145, miR-181c, miR199a and miR-1183 were hyper-expressed in the leukocyte fraction from the blood of patients with MTLE, and these can be potential biomarkers to aid in the diagnosis. We found no statistical differences of these miRNAs in blood and hippocampus of patients Engel I versus Engel III and IV, however, miR-1183 was progressively expressed in the blood of patients in the control groups, Engel I and Engel III and IV respectively.

Introdução: A recorrência de crises epiléticas pós cirurgia para tratamento da epilepsia do lobo temporal tem sido classificada como precoce ou tardia dependendo do tempo da primeira crise depois do procedimento. Contudo, este tempo de recorrência é variado, sendo arbitrariamente definido nos artigos científicos. Objetivos: Nós desenvolvemos um modelo matemático que pode identificar pacientes com chance de recorrência de crises pós-operatórias precoces ou tardias. Após, analisamos os dois grupos para identificar as diferenças clínicas, eletrofisiológicas e de neuroimagem entre estes pacientes. Métodos: Uma coorte histórica com 247 pacientes tratados com cirurgia para epilepsia do lobo temporal foi estudada. Dentre os paciente onde as crises epiléticas retornaram, utilizamos o tempo de recorrência em uma curva ROC para avaliar com maior acurácia o melhor período para predizer o prognóstico cirúrgico a longo prazo. Com isto, dividimos os pacientes em dois grupos: os de recorrência precoce e os de recorrência tardia. Após, nós analisamos as diferenças clínicas e radiológicas entre estes pacientes. Resultados: As crises epiléticas retornaram em 107 (48.9%) pacientes. A curva ROC mostrou que 6 meses era o tempo onde o prognóstico a longo prazo poderia ser determinado com maior acurácia (AUC = 0.761; sensibilidade = 78.8%; especificidade = 72.1%). Nós observamos que nos pacientes onde a recorrência ocorreu nos primeiros 6 meses após a cirurgia, a epilepsia começou mais precocemente (OR: 6.034; CI95%: 1.056–11.013; p=0.018), estes pacientes possuíam um pior prognóstico (6.849;2.538–18.518;p=0.001), necessitaram de maior número de medicação anti-epilética após o procedimento (2.07;1.162–9.345;p=0.013) e mais freqüentemente foram submetidos a uma nova cirurgia para controle de crises (9.592;1.181–77.877;p=0.021). Nos pacientes com recorrência tardia, as crises epiléticas eram mais comumente associadas a fatores desencadeantes (9.615;3.521–26.315;p<001). Conclusão: Pacientes com recorrências de crises epiléticas precoces ou tardias possuem diferentes características que podem estar relacionadas a diferenças entre os tipos de zonas epileptogênicas, eficácia dos procedimentos cirúrgicos ou mesmo à própria epileptogenicidade. A existência de tais disparidades podem ajudar a explicar os diferentes padrões de recorrência de crises pós cirurgia para epilepsia, auxiliando no planejamento do tratamento destes pacientes a longo prazo. / Background: Recurrence of seizures after surgery for epilepsy has been generally classified as either early or late depending on the time between surgery and seizure recurrence. However, the time of seizure recurrence is variable and it has been arbitrarily defined in the literatures. Objective: Here we establish a statistical-based model for discriminating patients with early or late seizure recurrence and examine the clinical, electrophysiological, and neuroimaging differences between these two groups of patients. Methods: A historical cohort of 247 patients treated surgically for temporal lobe epilepsy was identified. In those patients who recurred, the post-operative time until seizure recurrence was examined using an ROC curve to discriminate with greater accuracy the best period for predicting the long-term prognosis of patients. This approach divided patients in two groups, those with early and those with late seizure recurrence. Following this division, we compared differences between these groups in terms of a number of clinical and radiological variables. Results: Seizures recurred in 107 (48.9%) patients. The ROC curve showed that 6 months was the time when long-term surgical outcome could be determined with best accuracy, (AUC = 0.761; sensitivity = 78.8%; specificity = 72.1%). We observed that patients with seizure recurrence during first 6 months after surgery started seizing at younger age (OR: 6.034; CI95%: 1.056–11.013; p=0.018), had a worse outcome (6.849;2.538–18.518;p=0.001), needed a higher number of antiepileptic medications after surgery (2.07;1.162–9.345;p=0.013) and were more frequently submitted to reoperations (9.592;1.181–77.877;p=0.021). Patients with late relapse more frequently had seizures associated with trigger events (9.615;3.521–26.315;p<001). Conclusion: Patients with early or late recurrence of seizures have different characteristics that might reflect diversity in the epileptogenic zone and epileptogenicity itself. These disparities might help to explain variable patterns of seizures recurrence after epilepsy surgery.

Epilepsy is a neurological disorder that is characterized by aberrant electrical activity in the brain resulting in at least two unprovoked seizures over a period longer than 24 hours. Approximately 60% of individuals with epilepsy are diagnosed with temporal lobe epilepsy (TLE) and about one third of those individuals do not respond well to anti-seizure medications. This places those individuals at high risk for sudden unexpected death in epilepsy (SUDEP). SUDEP is defined as when an individual with epilepsy, who is otherwise healthy, dies suddenly and unexpectedly for unknown reasons. SUDEP is one of the leading causes of death in individuals with acquired epilepsies (i.e. not due to genetic mutations), such as TLE. Previous studies utilizing genetic models of epilepsy have suggested that circuitry within the vagal complex of the brainstem may play a role in SUDEP risk. Gamma-aminobutyric acid (GABA) neurons of the nucleus tractus solitarius (NTS) within the vagal complex receive, filter, and modulate cardiorespiratory information from the vagus nerve. GABAergic NTS neurons then project to cardiac vagal motor neurons, eventually effecting parasympathetic output to the periphery. In this study, a mouse model of TLE was used to assess the effect of epileptogenesis on GABAergic NTS neuron function and determine if functional alterations in these neurons impact SUDEP risk. It was discovered that mice with TLE (i.e. TLE mice) have significantly increased mortality rates compared to control animals, suggesting that SUDEP occurs in this model. Using whole cell electrophysiology synaptic and intrinsic properties of GABAergic NTS neurons were investigated in TLE and control mice. Results suggest that during epileptogenesis, GABAergic NTS neurons become hyperexcitable, potentially due to a reduction in A-type potassium channel current and increased excitatory synaptic input. Increases in hyperexcitability have been shown to be associated with an increased risk of spreading depolarization and action potential inactivation leading to neuronal quiescence. This may lead to a decreased inhibition of parasympathetic tone, causing cardiorespiratory collapse and SUDEP in TLE.

Purpose: Excessive day time sleepiness, sleep disorders and neurobehavior changes are common clinical observations in the patients with epilepsy. From literature review, they were highly related with epilepsy risk characteristics (age of onset, types or numbers of drugs, seizure frequency), co-morbidities or neuronal network changes. The serological biomarkers have been reported to reflect the phenomenon of seizure, while their correlations with neurobehavior changes were still not concluded. There were two purposes of this thesis. (1) To understand the relationship between sleep disturbance with day time performances (2) To understand the relationships between serological biomarkers, neurobehavior performances and neuronal networks in patients with temporal lobe epilepsy. Material and Methods: The study enrolled patients from epilepsy outpatient clinic. By using self-appreciated questionnaire (The Pittsburgh Sleep Quality Index, The Epworth Sleepiness Scale, Euroqol Quality of Life Scale-5D), we collected the characteristics of sleep related behavior and life quality changes and explored the relationship with epilepsy risk characteristics. In patients with temporal lobe epilepsy, we assessed the neurobehavior performances, measured the serological biomarkers (heat shock protein 70, S100£]protein, neuron specific enolase, brain derived neurotrophic factor, plasma and mitochondrial DNA) and brain magnetic resonance imaging. In statistical analysis, we compared the differences with age matched controls or performed correlation analysis among the parameters Result: One hundred and seventeen patients with epilepsy completed the sleep quality questionnaires. The results showed that 20 percent of patients had day time sleepiness, while the sleep disorder was prolong sleep latency and impaired sleep efficiency. In epilepsy characteristics, patients with complex partial seizure, intractable seizure or with multi-pharmacy were related with poor sleep quality. A total of 34 patients completed the serological, neurobehavior and brain magnetic resonance analysis. The results showed that patients with temporal lobe epilepsy had higher heat shock protein 70 and S100£]protein levels, while those with attacks more than twice per month had significant higher heat shock protein 70, S100£]protein and neuron specific enolase levels. Compared with the matched controls, the regions showing atrophy included hippocampus and parahippocampus, putamen, thalamus and supplementary motor areas. In correlation study, only heat shock protein 70 showed an inverse correlation with hippocampal volume (R square = 0.22, p = 0.007) after controlling for the effect of age. Conclusion: The study suggested that epilepsy risk characteristics, serological biomarkers, brain atrophic regions were important factors for day time sleepiness, sleep disturbances and neurobehavior changes in patients with epilepsy.

The aim of the thesis was to conduct a detailed examination of the evidence pertaining to the cerebral lateralisation of musical function. Theoretical models from the neuropsychological and cognitive psychology fields were employed, with emphasis placed on the way the models interrelate to gain a more coherent account of music cognition and its relationship to cerebral lateralisation. (For complete abstract open document.)

Thesis (doctoral)--Westfälischen Wilhelms-Universität zu Münster, 1995. / Includes bibliographical references.

Thesis (D.Phil (Social Work and Criminology))--University of Pretoria, 2005. / Includes abstract in English and Afrikaans. Bibliography.