<p> Animal studies have shown that the tonotopic organization of the auditory cortex is not statically fixed, but can be remodeled by experience. The purpose of this study was to investigate whether or not frequency discrimination training can induce changes in the cortical representation of a selected frequency in humans. Six human subjects were trained for approximately 3 weeks to detect a change in pitch between two tones (40Hz amplitude modulated) using a standard frequency of 2040 Hz. Each subject was tested on his/her discriminative ability before and after training using three different standards (2040Hz, 1840Hz, and 2240Hz). EEG data were recorded both before and after training and changes in transient and steady-state responses were investigated. Behaviourally, every subject improved at the discrimination task using the trained frequency. However, only three subjects demonstrated transfer to both untrained frequencies. In the EEG data, the P2-Nl amplitude increased in five of the six subjects and the Nllatency decreased in all six for the 2040Hz set. These two findings were statistically significant (p<0.05) for the group. There were no statistically significant findings for the side frequencies. The change in the 40 Hz steady-state response was also not significant, increasing in three subjects and decreasing in the other three. These findings indicate that changes are expressed in the secondary auditory cortex. These findings may also be applicable to the treatment of tinnitus. </p> / Thesis / Master of Science (MSc)

Des mutations du gène ARX (aristaless-related homeobox gene) ont été identifiées dans un large spectre de désordres neurologiques précoces, incluant ou non des malformations cérébrales, le plus souvent associés à des épilepsies. Il est proposé que le gène ARX, codant pour un facteur de transcription, joue un rôle primordial au cours du développement cérébral, notamment sur la migration des neurones GABAergiques, mais son implication au cours de la mise en place du système nerveux central reste cependant encore mal connue. L’objectif de ce travail a été d’étudier le rôle du gène ARX et les conséquences de ses mutations sur le développement cérébral dans le but de mieux comprendre ces pathologies. Dans un premier temps, nous avons étudié l’effet d’une mutation particulière du gène, la mutation ARX(CGC)7, une expansion polyalanine retrouvée principalement dans des pathologies sans malformation cérébrale mais avec des épilepsies, tels que les syndromes de West ou d’Ohtahara. Des analyses réalisées sur une lignée de souris knock-in pour cette mutation (GCG)7 et sur des rats après électroporation in utero ont montré que la migration neuronale des neurones glutamatergiques et GABAergiques ainsi que la maturation des neurones GABAergiques ne sont pas altérées par cette mutation. De façon intéressante, nos données suggèrent que les épilepsies observées chez les souris knock-in résulteraient plutôt d’une réorganisation du réseau glutamatergique. Etant donné que le gène ARX n’est pas exprimé dans les neurones glutamatergiques, l’ensemble de ce travail suggère donc que les épilepsies chez les souris knock-in pour la mutation (GCG)7 sont la conséquence d’une altération développementale secondaire à la mutation initiale du gène, et ceci aurait d’importantes répercussions thérapeutiques qui requièrent d’avantages d’études. Des expériences nous ont ensuite permis d’étudier l’effet de plusieurs mutations du gène ARX sur la morphologie des interneurones in vitro. Celles-ci ont montré que les mutations d’ARX n’engendrent pas une localisation subcellulaire anormale de la protéine dans les interneurones en culture. De façon intéressante, ces expériences suggèrent que la morphologie des interneurones est altérée seulement par certaines mutations, notamment les mutations P353R et Dup24. Ces données soulignent ainsi l’importance d’étudier de façon spécifique chaque mutation du gène pour expliquer les mécanismes engendrant l’hétérogénéité phénotypique liée aux mutations d’ARX. L’ensemble de ces travaux contribuent à une meilleure compréhension du rôle du gène ARX dans le développement cortical et à une meilleure caractérisation des mécanismes physiopathologiques des désordres neurologiques précoces liés aux mutations de ce gène. / Several mutations in ARX gene (aristaless-related homeobox gene) have been found in a large spectrum of infantile neurological disorders, with or without cerebral malformation, but frequently linked to epilepsy. It has been proposed that ARX, coding for a transcription factor, plays a crucial role in brain development, especially in migrating interneurons, but its involvement in nervous system development still remains to be clarified. The aim of this work has been to study the role of ARX gene and the consequences of ARX mutations on cerebral development in order to better understand these pathologies.We have first investigated the effects of an ARX polyalanine expansion, the mutation (GCG)7, which was found in pathologies without brain malformation but associated to epilepsy, such as West and Ohtahara syndromes. Analysis performed on knock-in mice for this mutation and in utero electroporated rat brains have shown that this mutation doesn’t alter neither glutamatergic and GABAergic neuronal migration, nor GABAergic neuron maturation. Interestingly, our data suggest that epilepsy observed in knock-in mice would result rather from a reorganization of glutamatergic networks. Since ARX gene is not expressed in excitatory neurons, our work suggests that epilepsy observed in knock-in mice is the consequence of developmental alterations secondary to the initial mutation, and this would have crucial therapeutic implications that require additional investigations. In vitro experiments have then allowed us to study the effect of several ARX mutations on interneurons morphology. These experiments have shown no abnormal subcellular localization of ARX protein following transfection of these different mutations in cultured interneurons. Interestingly, our data show that interneuron morphology is altered only by some mutations, particularly the P353R and the Dup24 ARX mutations. Our data underline the importance to study specifically each mutation in order to explain mechanisms generating phenotypic heterogeneity linked to ARX mutations.Taken together, this study contributes to a better understanding of ARX involvement in cerebral development and to a better characterization of pathophysiological mechanisms linked to ARX mutations.

The human brain changes in a complex and heterogeneous way throughout life, the normal aging process is associated to significant alterations in the axonal connections. In this study, we evaluated the age-related changes in physical parameters associated with the brain white and gray matter integrity in healthy subjects, as well as the possible correlation between them in specific tracts. Structural images (1 mm isotropic) and diffusion weighted images (2 mm isotropic, b = 1000 s /mm2) of 158 healthy individuals aged between 18 and 83 years were retrospectively collected at the Clinics Hospital of Ribeirao Preto, after their acquisition in a 3T MR scanner. From the structural images, the cortical thickness was estimated and the age effect was evaluated in several regions based on the Atlas of Destrieux. The diffusion-weighted images were processed to characterize the intravoxel diffusion using two models: diffusion tensor (DT) and constrained spherical deconvolution (CSD). Fractional anisotropy (FA) and apparent density of fiber (AFD) maps were estimated and used in statistical group analysis between the three groups separated by age. The most relevant brain tracts were segmented by three procedures: manually, automatically with a specific tool and based on automatic segmented cortical regions. Physical parameters of diffusion (anisotropy and diffusivities) were evaluated in the segmented tracts to determine the age-related changes. The connectome analysis based on two cortical parcellations was performed to evaluate the age effect on characteristic structural brain network parameters. The tract-cortical relationship was evaluated considering the anisotropy of each tract and the thickness of the cortical areas at the end of the corresponding tract. Further analysis was performed to evaluate a possible association of structural and functional connectivity in the corpus callosum (CC). There was significant cortical thinning in 88.5% of the regions during life (p <0.05, corrected for multiple comparisons); the frontal region was the most affected in the initial aging (after 40 years), and the occipital and temporal regions in the elderly (after 60 years). Similarly, the group analysis demonstrated a global pattern of reduction of FA and AFD in the white matter, with a higher rate of degradation of integrity from the sixth decade of life. The manual selection of tracts from the DT model proved to be the most reliable methodology in the precise definition of the tracts for our data. Following this methodology, analysis of anisotropy and diffusion parameters also indicated degeneration of white matter in normal aging in all studied brain tracts and corroborated to the antero-posterior gradient of degeneration in the CC. Fornix was the most affected tract bilaterally, with a 3.5% reduction and an increase of 4% per decade in these parameters, respectively; followed by CC. In the evaluation of the age effect on the connectome estimates, regardless of diffusion model and cortical atlas, there was a decrease in global efficiency, number of connections and local efficiency with aging, mainly in the prefrontal, temporal and parietal and its connections. In the tract-cortical analysis, cortical regions connected by tracts demonstrated similar thinning patterns for the majority of tracts, and a significant relation between mean cortical thinning rate and FA/MD alteration rates were found. In all evaluated tracts, age was the main effect controlling diffusion parameters alterations; there were no direct correlations with cortical thickness for the majority of tracts. Only for the fornix, the values of FA and MD showed significant correlation with the cortical thickness of the subcallosal gyrus in both hemispheres during aging (p <0.05 corrected). For the other tracts, CC, Inferior Longitudinal Fasciculus, Uncinated Fasciculus, Inferior Fronto-occipital Fasciculus, Corticospinal Tract, Cingulum and Arcuate Fasciculus, age was the main effect controlling alterations in the parameters, but there were no direct correlations between FA and MD and cortical thickness during the aging process. / O cérebro humano muda de forma complexa e heterogênea ao longo da vida, o processo de envelhecimento normal tem associado significativas alterações nas conexões axonais. Neste estudo, avaliamos as mudanças relacionadas à idade em parâmetros físicos associados à integridade das substâncias branca e cinzenta cerebral em sujeitos saudáveis, assim como a possivel correlação entre eles em tratos específicos. Imagens estruturais (1 mm isotrópica) e imagens ponderadas em difusão (2 mm isotrópica e b=1000 s/mm2) de 158 indivíduos saudáveis entre 18 a 83 anos foram coletadas retrospectivamente no Hospital das Clinícas de Ribeirão Preto, após sua aquisição em aparelho de ressonância magnética de 3 Teslas. A partir das imagens estruturais, a espessura cortical foi estimada e o efeito de idade nela foi avaliado em diversas regiões tomando com base o atlas de Destrieux. As imagens ponderadas em difusão foram processadas para caracterizar a difusão intravoxel utilizando dois modelos: tensor de difusão (DT) e deconvolução esférica restrita (CSD). Mapas de anisotropia fracionada (FA) e densidade aparente da fibra (AFD) foram estimados e usados em analise estatistica de três grupos separados por faixa etária. Os tratos cerebrais mais relevantes foram segmentados por tres procedimentos: manualmente, automaticamente com uma ferramenta especifica e com base em regiões corticais automaticante segmentadas. Parâmetros físicos de difusão (anisotropia e difusibilide) foram avaliados nos tratos segmentados para determinar as alterações relacionadas à idade. A análise de conectoma baseada em dois parcelamentos corticais foi realizada para avaliar também o efeito da idade em parâmetros caracteristicos da rede estrutural cerebral. A relação trato-cortical foi avaliada considerando a anisotropia de cada trato e as espessuras das áreas corticais nas extremidades do trato correspondente. Uma análise adicional foi realizada para avaliar uma possivel associação de onetividades estrutural e funcional no corpo caloso (CC). Houve afinamento cortical significativo em 88,5% das regiões durante a vida (p <0,05, corrigido); a região frontal foi a mais afetada no envelhecimento inicial (após 40 anos), e as regiões occipital e temporal nos idosos (após 60 anos). Similarmente, a análise de grupo demonstrou um padrão global de redução de FA e AFD na substância branca, com uma maior taxa de degradação de integridade a partir da sexta década de vida. A seleção manual de tratos baseada no modelo de DT mostrou-se a metodologia mais confiavél na precisa definição dos tratos nos nossos dados. Seguindo essa metodologia, a análise dos parâmetros de anistropia e difusão também indicou degeneração de substância branca no envelhecimento normal em todos os tratos cerebrais estudados e corroborou o gradiente ântero-posterior de degeneração no CC. O fornix foi o trato mais afetado bilatreamente com redução de 3.5% e aumento de 4% por década nesses parâmetros, respectivamente; seguido do CC. Na avaliação do efeito da idade nas estimativas do conectoma, independentemente do modelo de difusão e do atlas cortical usado, houve uma diminuição da eficiência global com o envelhececimento, do número de conexões e da eficiência local, principalmente nas regiões pré-frontal, temporal e parietal e suas conexões. Nas análises trato-corticais, as regiões corticais conectadas por tratos mostraram padrões de afinamento similares para a maioria dos tratos, e uma correlação significativa entre a taxa média de afinamento cortical e as taxas de alteração de FA e difusibilidade média (MD) foram encontradas. Em todos os tratos avaliados, a idade foi o principal efeito controlando das alterações dos parâmetros de difusão; não houve correlações diretas com espessura cortical para a maioria dos tratos. Somente para o fornix, os valores de FA e MD mostraram correlação com a espessura cortical do giro subcalosal (parcelamento de Destrieux) em ambos os hemisférios durante o envelhecimento (p <0,05 corrigido). Para os outros tratos, CC, fascículo longitudinal inferior, fascículo uncinado, fascículo occipitofrontal inferior, trato cortico-espinal, parte cingulada do cíngulo e fascículo arqueado, a idade foi o principal efeito no controle das alterações dos parâmetros, mas não houve correlações diretas entre FA e MD e espessura cortical durante o processo de envelhecimento

Cette thèse concerne la segmentation d’images HRpQCT et l’évaluation d’indices morphologiques de l’os cortical pour le diagnostic de l’ostéoporose et la prédiction des fractures osseuses. Dans un premier temps,deux méthodes sont proposées pour la segmentation de l’os cortical. La première utilise une nouvelle approche des contours actifs basée sur la logique floue suivie d’une nouvelle technique de remplissage développée pour imiter le comportement des opérateurs pour séparer l’os cortical de l’os trabéculaire. La deuxième approche est une technique 3D à double contours actifs combinant à la fois les informations locales le long et entre les deux contours. Les deux approches de segmentation sont comparées à celles de l’état de l’art afin de valider leurs performances. Dans un second temps, différents indices extraits de l’os cortical sont utilisés pour déterminer leur potentiel de prédiction des fractures ostéoporotiques. Les résultats obtenus montent que l’analyse globale de l’os cortical masque des variations potentiellement importantes.Par conséquent, une décomposition régionale de l’enveloppe corticale est proposée afin d’améliorer la prédiction du risque fracturaire. / This thesis concerns the segmentation of HRpQCT images and the evaluation of the cortical bone parameters for the osteoporosis characterization and the fracture prediction. Firstly, two approaches were proposed to segment the cortical bone. The first uses a new fuzzy energy active contours approach followed by a new filling technique designed to mimic the behaviour of clinicians while extracting the cortical bone from the trabecularone. The second approach is a local based 3D dual active contours approach proposed to separate between three regions constituting the image. To move, this approach combines the local information along each point in the two contours conjointly with the information between them. The segmentation results of these approaches were confronted to the state of the art methods to validate their performance. Secondly,different parameters were extracted from the segmented cortical bone to monitor the association of these parameters with the osteoporotic fracture prediction. Global analysis of the cortical bone obscures potentially important regional variations. Therefore, regional cortical decomposition was proposed to illustrate that cortical sub-regions could improve the evaluation of fracture risk than the global analysis of the cortical bone.

A literatura recente sugere que a fadiga mental piora o desempenho de atletas durante testes contrarrelógio (TT) de ciclismo. Assim o objetivo deste estudo foi verificar se a cafeína poderia reverter ou bloquear os efeitos negativos da fadiga mental em um contrarrelógio de 20km (TT20km). Métodos: Doze ciclistas treinados, com experiência em TT há pelo menos 2 anos, e nível de performance > 3 (34.3 ± 6.2 anos; 179.3 ± 5.1 cm; 77.6 ± 6.8 kg; 13.8 ± 4.5 % de gordura corporal; 58.9 ± 6.2 ml.kg-1.min-1; 367.0 ± 32.5 WPEAK), realizaram cinco visitas ao laboratório: 1) PRELIMINAR: para seleção dos sujeitos e familiarização com os instrumentos; 2) CONTROLE: foram realizadas três contrações voluntárias isométrica máxima (CVM), cujo maior torque encontrado entre elas definirá o valor de 70% para contração voluntária isométrica submáxima (CVIS) nas sessões seguintes. Posteriormente a CVIS, foi realizado o TT20km sem manipulação cognitiva ou suplementação; 3) FADIGA MENTAL (FM): execução do teste de atenção sustentada (RVIP), CVIS e TT20km respectivamente; 4) PLACEBO: ingestão da cápsula de placebo (FM + PLA) em seguida teste de RVIP, CVIS e TT20km; 5) CAFEÍNA: ingestão de cafeína (FM + CAF), consequentemente realizaram o teste de RVIP, CVIS e o TT20km). A atividade cerebral do córtex pré-frontal e córtex motor primário foi analisada por meio de eletroencefalografia (EEG) registrada antes e depois do teste RVIP. Igualmente, EEG do córtex pré-frontal e motor primário foi registrada durante uma contração voluntária isométrica submáxima, realizada antes do TT20km. Para as análises de comparações entre pré e pós tarefa cognitiva de atenção sustentada utilizou-se um teste T-student. Para as comparações múltiplas analisou-se por modelo misto entre a condição e a distância no TT20km, quando necessário o teste de Bonferroni foi utilizado. Resultados: CAF melhorou o desempenho em ciclistas mentalmente fatigados em ~ 1.8 % para FM e ~ 1.7 % para PLA (p = 0.00) e aumentou ~ 4.4 % para FM e ~ 3.6 % do que o placebo de WMEAN em FM + CAF (p = 0,00). A ativação cortical aumentou em ~ 4.8 % nas ondas teta de EEG após teste RVIP em FM e reduziu em FM + CAF ~ 8.8 % e FM + PLA ~ 4.8%. A razão entre PSE e WMEAN foi menor em FM + CAF do que nas outras condições (p = 0.01). Os ciclistas apresentaram maior motivação nos primeiros 2 km em FM + CAF comparado a FM e FM + PLA (p = 0.02) e maior afeto nos últimos 2 km em FM e FM + CAF do que FM + PLA (p = 0.01). Uma menor ativação cortical durante a CVIS e uma maior eficiência neuromuscular foi identificada na condição FM + CAF (p = 0.03). Conclusão: A CAF bloqueou os efeitos negativos da fadiga mental no CPF, melhorou o desempenho, diminuiu a PSE durante TT20km e aumentou a eficiência neuromuscular / Recent literature suggests that mental fatigue worsens the performance of athletes during time trial (TT) cycling tests. Thus, the aim of this study was to verify if caffeine could reverse or block the negative effects of mental fatigue in a TT of 20km (TT20km). Methods: Twelve trained cyclists with experience in TT for at least 2 years, and performance level > 3 (34.3 ± 6.2 years, 179.3 ± 5.1 cm, 77.6 ± 6.8 kg, 13.8 ± 4.5 % body fat 58.9 ± 6.2 ml .kg-1.min-1; 367.0 ± 32.5 WPEAK), carried out five visits at laboratory: 1) PRELIMINARY: for selection of subjects and familiarization with the instruments; 2) CONTROL: three maximal isometric voluntary contractions (CVM) were performed, the highest torque found between them will define the value of 70% for voluntary isometric submaximal contraction (CVIS) in the following sessions. After the CVIS, the TT20km will be performed without cognitive manipulation or supplementation; 3) MENTAL FATIGUE (FM): execution of the sustained attention test (RVIP), CVIS and TT20km respectively; 4) PLACEBO: placebo capsule intake (FM + PLA) then RVIP, CVIS and TT20km test; 5) CAFFEINE: caffeine intake (FM + CAF), consequently performed the RVIP, CVIS and TT20km tests). Brain activity of the prefrontal cortex and primary motor cortex will be analyzed by electroencephalography (EEG) recorded before and after the RVIP test. Likewise, EEG of the prefrontal cortex and primary motor will be recorded during a submaximal isometric voluntary contraction, performed before TT20km. For the analysis of comparisons between pre and post cognitive task of sustained attention was used a T-student test. For the multiple comparisons, the mixed model was analyzed for the condition and the distance in the TT20km, and the best matrix was identified for each case, when necessary the Bonferroni test was used. Results: CAF improved performance in mentally fatigued cyclists at ~ 1.8 % for FM and ~ 1.7 % for PLA (p = 0.00) and increased ~ 4.4% for FM and ~ 3.6% than WMEAN on FM + CAF (p = 0.00). Cortical activation increased by ~ 4.8 % in the theta EEG waves after RVIP test in FM and reduced in FM + CAF ~ 8.8 % and FM + PLA ~ 4.8 %. The ratio of RPE to WMEAN was lower in FM + CAF than in other conditions (p = 0.01). The ciclists started the TT20km more motivated in first 2 km (p = 0.02) and finished with more pleasure in the last 2km in FM and FM + CAF than in FM + PLA (p = 0.01). Lower cortical activation during CVIS and greater neuromuscular efficiency was identified in the FM + CAF condition (p = 0.03). Conclusion: CAF blocked the negative effects of mental fatigue on CPF, improved performance, decreased PSE over TT20km and increased neuromuscular efficiency

Este trabalho teve como objetivo estudar um caso específico de epilepsia refratária causada por uma má formação no tecido cerebral, denominada displasia cortical focal (DCF). Por ser uma má formação no cérebro, suas consequências aparecem desde a infância, em que ela, a DCF, é a principal causadora das epilepsias de caso refratário. O mapeamento da região com DCF geralmente é feito por meio de imagens de ressonância magnética em conjunto com outras técnicas, como, por exemplo, o PET (positron emission tomography), o EEG (eletroencefalograma) intracraniano, entre outras. Contudo, por serem técnicas muito caras, de difícil realização ou muito invasivas, e por sabermos que as regiões displásicas possuem uma rigidez diferente da do restante do cérebro, foi proposto nesta dissertação o estudo desses casos utilizando uma técnica barata, simples, não invasiva e sensível à rigidez tecidual, a elastografia por ultrassom, na qual, para causar a deformação do tecido cerebral, foram usadas próprias artérias internas do cérebro. Para tal estudo, criamos um algoritmo de processamento de dados com uma interface gráfica GUI (graphical user interface) capaz de mudar os parâmetros de processamento e ver seus resultados em tempo real. Em seguida, esse algoritmo foi estudado em um ambiente controlado em material mimetizador de tecido biológico (phantom), no qual construímos um bloco de 10 x 10 x 12cm3, preenchido com material que mimetiza as propriedades mecânicas e acústicas do tecido mole e inserimos nele uma bexiga canudo preenchida com um uido simulador de sangue e uma inclusão mais rígida do que a base do material, posicionada acima do canudo. Foi utilizado, também, um acionador mecânico pulsátil para simular a pulsação mecânica equivalente à pulsação sanguínea da artéria cerebral. Foram feitas imagens elastográcas e de velocidade utilizando somente a deformação causada pelo deslocamento da bexiga, no interior do phantom, e, através de uma transformada de Fourier, foi calculado o período de pulsação da bexiga. Vimos que as imagens elastográcas e de velocidade foram capazes de localizar a inclusão, e o processamento temporal pode nos mostrar com precisão a frequência de pulsação da bexiga canudo. Finalizada essa etapa laboratorial, zemos o mesmo procedimento, porém in vivo, para dois casos: um com DCF tipo III-B, no qual não enxergávamos nada no modo B; e outro com tipo II-B, no qual foi observado uma diferença de impedância mecânica pelo modo B. As imagens foram coletadas durante o procedimento cirúrgico pelo próprio cirurgião usando um transdutor microconvexo acoplado a uma plataforma de ultrassom, modelo Sonix RP, e processadas num segundo momento. Vimos, no primeiro caso, pelas imagens elastográcas, as regiões mais rígidas, supostamente displásicas, que não estavam aparecendo no modo B e, no segundo caso, uma região maior do que a apresentada no modo B. Nossos resultados das medidas de frequência da pulsação arterial, para ambas as situações, 61; 5BPM e 91BPM, caram bastante próximos do valor medido com o eletrocardiograma durante a coleta do sinal, 65BPM e 94BPM, respectivamente. Por meio dos resultados da análise histológica, pudemos conrmar que o que estávamos enxergando com nosso programa era realmente uma região displásica. Dessa forma, concluímos que nosso algoritmo funcionou bem para esses casos clínicos. / The mainly goal of this work was to study a specic case of refractory epilepsy generated by a malformation in the brain tissue, called focal cortical dysplasia (FCD). Due the fact it is a brain malformation its eects show up since the childhood where it is the principal epilepsy generator. The mapping of this region is usually made by magnetic resonance images with another technique, such as, for instance, the PET (position emition tomography), the EEG (electrocardiogram), and others. However, for the fact that these techniques are expensive, dicult to perform or invasive, and knowing that the dysplastic regions are stier than the regular brain tissue, it was proposed in this dissertation the use of ultrasound elastography as a cheaper, simpler and noninvasive image modality capable to detect dierences in the tissue stiness of the FCD region. To generate the strain in the brain tissue it was used the pulsation of the local arteries. To achieve our goal, we created a data processing algorithm in MATLAB with a graphic user interface (GUI) capable to change the processing parameters to see its results in real time. This algorithm was tested in phantom using a block of tissue mimicking material (10 x 10 x 12 cm3). A balloon of latex led with a blood mimicking uid was immersed in the middle of the phantom and a cylindrical inclusion of 1 cm of diameter was immersed above the balloon. The bulb of the balloon was keep outside of the phantom to be mechanically pressured by a dedicated magnetic actuator, simulating the mechanical pulsation of the brain arteries. The velocity and elastography images were studied using just the strain caused by the displacement of the wall of the balloon tube inside the phantom. The period of pulsation was precisely calculated from these images. After that, we did the same procedure in two in vivo cases: one with FCD type III-B; and the other with FCD type II-B. All our intraoperative images were acquired for the surgeons using a micro convex transducer linked to an Ultrasound platform (Sonix RP) and, then, processed o-line. In the B mode scanning we didnât see any formation inside the brain for the rst case, and for the second, we did. In the elastographic images we saw a clearly stiffer region in the rst case that was invisible in the B mode; and for the second case, we saw a bigger stiffer region than we saw in the B mode imaging too. And for both results, the arteria pulsation frequency, 61.5 BPM and 91 BPM, were veryclose to the measured value collected in the electrocardiogram during the surgery, 65 BPM and 94 BPM, respectively. Analyzing the histological results we could conrm that what we were showing in our elastographic images were FCD, indeed. Thereby we concluded that our algorithm had worked in these clinical data.

As displasias corticais focais constituem um grupo de malformações do desenvolvimento cortical cerebral. São consideradas a causa mais comum de epilepsia refratária na população pediátrica. A hemimegalencefalia faz parte deste grupo de malformação do desenvolvimento cortical e clinicamente devastadora em crianças, caracterizada pelo crescimento distorcido e anormal de um hemisfério cerebral. O mTOR (Mammalian Target of Rapamycin) é uma proteína quinase, que normalmente funciona como um regulador central de importantes funções fisiológicas, incluindo o crescimento e proliferação celular, metabolismo, autofagia, e sobrevivência e morte celular. O objetivo deste estudo foi identificar o defeito genético específico da hemimegalencefalia analisando os genes das vias de sinalização do mTOR. Foram selecionados 10 pacientes diagnosticados com hemimegalencefalia com faixa etária entre 0 á 18 anos de idade. Os pacientes submetidos ao procedimento cirúrgico foram designados para a coleta do material biológico (sangue e tecido encefálico) para análise genômica. Foram encontradas variantes somáticas de três genes relacionados a via mTOR com alto índice de patogenicidade, sendo elas; mutações missense na MTOR, HME 6584 (c.7255G> A, p.Glu2419Lys), HME 4146 (c.7498A> T, p.L7105f) mutações missense do gene PIK3CA, HME 4149 E542K (c.1624G> A), HME 4143 (c.1258T> C, p.C420R). A hipótese mediante esses resultados é que a mutação somática de genes que estão presentes na via mTOR podem ser uma das causas genéticas da HME. Essas observações sugeriram que a HME representa um espectro de distúrbios do neurodesenvolvimento resultando de distintas progenitoras que são determinados pelo tempo em que a mutação ocorreu durante o desenvolvimento cerebral. Pode-se esperar que uma mutação que ocorre precocemente durante o desenvolvimento afete um grande número de células e resulte em uma malformação maior, ao passo que a mesma mutação ocorrendo mais tarde no desenvolvimento poderia causar uma menor malformação. No futuro, numerosas mutações somáticas em genes conhecidos ou novos serão, sem dúvida, reveladas em amostras de cérebros ressecados e assim, possíveis correlações entre genótipos e fenótipos podem emergir, permitindo que o diagnóstico clínico genético ajude a prever o desfecho do paciente / Focal cortical dysplasias constitute a group of malformations of cerebral cortical development. They are considered the most common cause of refractory epilepsy in the pediatric population. Hemimegalencephaly is part of this group of malformation of cortical development and clinically devastating in children, characterized by the distorted and abnormal growth of a cerebral hemisphere. MTOR (Mammalian Target of Rapamycin) is a protein kinase, which normally functions as a central regulator of important physiological functions, including cell growth and proliferation, metabolism, autophagy, and cell death and survival. The aim of this study was to identify the specific genetic defect of hemimegalencephaly by analyzing the genes of the mTOR signaling pathways. Ten patients diagnosed with hemimegalencephaly with ages ranging from 0 to 18 years of age were selected. Patients submitted to the surgical procedure were assigned to the collection of biological material (blood and brain tissue) for genomic analysis. Somatic variants of three genes related to the mTOR pathway with high pathogenicity index were found; missense mutations in the MTOR, HME 6584 (c.7255G> A, p.Glu2419Lys), HME 4146 (c.7498A> T, p.Leu7105Phe) missense mutations of the PIK3CA gene, HME 4149 (c.1624G> A Glu542Lys), HME 4143 (c.1258 -> C, p.Cys420Arg). The hypothesis by these results is that the somatic mutation of genes that are present in the mTOR pathway may be one of the genetic causes of HME. These observations have suggested that HME represent a spectrum of neurodevelopmental disorders resulting from distinct progenitors that are determined by the time the mutation occurred during brain development. A mutation that occurs early in development may be expected to affect a large number of cells and result in a larger malformation, whereas the same mutation occurring later in development could cause a minor malformation. In the future, numerous somatic mutations in known or new genes will undoubtedly be revealed in samples of resected brains and thus, possible correlations between genotypes and phenotypes may emerge, allowing genetic clinical diagnosis to help predict the outcome of the patient

Este estudo investiga o Módulo de Young (Módulo de Elasticidade) em amostras de osso cortical oriundas de uma mesma região da tíbia bovina depois de submetidas a congelamentos e descongelamentos sucessivos. As amostras foram coletadas da face caudal do terço médio da diáfise de tíbias bovinas e depois submetidas a testes de flexão em três pontos e ensaios de ruptura, obedecendo à norma ASTMD790M86&#949;1. Nesse experimento, 20 pares de amostras de ossos corticais foram retirados de 20 tíbias bovinas, cada amostra medindo 2 mm x 4 mm x 40 mm, aproximadamente. Todos os testes foram realizados à temperatura controlada (21 graus Celsius), e as condições de ensaio foram as mesmas durante os testes. As amostras foram identificadas e divididas em dois grupos experimentais: Grupo A: as amostras foram conservadas (em sacos plásticos) a - 20 graus Celsius (submetidos a congelamentos e descongelamentos sucessivos por 50 vezes: uma vez ao dia durante 50 dias). Esses descongelamentos produziram 15 ensaios de flexão, sendo que o primeiro foi realizado com as amostras ainda frescas mantidas em geladeira a 4 graus Celsius. Após os 50 descongelamentos foram levados até a ruptura. Grupo B: as amostras foram conservadas frescas (em sacos plásticos) a 4 graus Celsius em geladeira aguardando o tempo para que os ensaios do grupo A fossem concluídos e, posteriormente fez-se o teste de ruptura. Importantes cuidados foram tomados. Depois de cada descongelamento, as amostras foram analisadas e comparadas com as amostras frescas tanto para os ensaios de flexão em três pontos como para os ensaios de ruptura. Observamos que o congelamento não alterou de modo significativo as propriedades mecânicas destes ossos, pois não foi encontrada nenhuma diferença significativa entre o comportamento dos dois grupos testados. Isso implica que, congelando e estocando mesmo por longos períodos, as propriedades mecânicas não se alteram tanto nos ensaios de flexão em três pontos como nos ensaios de ruptura. Com respeito às diferenças observadas no dia a dia, as alterações do primeiro dia até o último dia não foram significativas / The present study was conducted to investigate the Young Module (Elasticity Module) in cortical bone samples submitted to successive freezing and thawing and then assayed in flexion tests at three points and in rupture tests according to the norms of ASTMD790M86&#949;1. Twenty pairs of cortical bone samples, each measuring approximately 2 x 4 x 40 mm, were collected from the same region of the caudal surface of the middle third of the diaphysis of 20 bovine tibiae and divided into two experimental groups: Group A: consisting of 20 cortical bone samples stored at – 20 Celsius degrees in plastic bags and submitted to successive freezing and thawing 50 times once a day for 50 days. These thawing episodes produced 15 flexion assays. After the 50 thawing episodes, the samples were tested until rupture. Group B: consisting of 20 cortical bone samples stored at 4 Celsius degrees in plastic bags in a refrigerator until the time when the assays of group A were concluded and then submitted to the rupture test. No statistically significant variation in the Young Module was observed after the flexion test at three points performed for Group A. The values of rupture tension also did not differ significantly between groups A and B. We observed that freezing did not cause a significant change in the mechanical properties of these bones, indicating that freezing and storage of these bones even for long periods of time does not alter their mechanical properties in flexion tests at three points or in rupture assays

A osteoporose é um enfraquecimento progressivo dos ossos, que ficam cada vez mais sujeitos a fraturas. Embora possa acometer ambos os sexos, ela é mais freqüente nas mulheres após a menopausa devido à diminuição dos hormônios femininos, os estrógenos. É uma doença que avança lentamente, sem sintomas, geralmente sem ser percebida até que aconteça uma fratura. Uma vez instalada a osteoporose, tem que se evitar maior perda óssea para prevenir fraturas. O objetivo deste trabalho é analisar o quadro de instalação de osteopenia em ossos corticais de ratas ovariectomizadas. Foi utilizado um modelo de osteopenia em 29 ratas Wistar ovarectomizadas com massa corpórea de 250 gramas. Os animais foram divididos em 6 grupos experimentais e eutanaziados em períodos diferentes: o primeiro grupo foi eutanaziado 30 dias após a cirurgia, que corresponde ao tempo de início da osteopenia, e a partir daí, os demais grupos foram eutanaziados numa seqüência de 15 dias até o 105º dia. A avaliação do quadro de instalação foi realizada através de medidas antropométricas e de propriedades mecânicas dos fêmures (ensaio de flexão de três pontos). Cada um dos itens obtidos foram comparados utilizando-se o programa GraphPad InStat 3. O teste t-Student foi aplicado para checar a variação do peso corporal com p < 0,05. Aplicaram-se os testes ANOVA e Student-Newmam-Keuls com coeficiente de variação também menor que 5% para os demais itens analisados. Observou-se um aumento significativo no comprimento dos fêmures durante o primeiro mês de experimento, assim como na carga máxima aplicada. Os resultados obtidos neste estudo sugerem que a ovariectomia é um fator que não causa grandes alterações mecânicas e geométricas na região cortical dos fêmures de ratas no período analisado / Osteoporosis is a progressive weakness of bones, which are more and more subjects to fractures. Although it happens to men and women, it is more frequent in postmenopausal women due to a decrease of female hormones. It is a disease without symptoms and it is usually noticed only after a fracture. Once the osteoporosis is installed, a bigger bone loss must be avoided in order to prevent a fracture. This study aims to analyze the osteopenia installation process in cortical bones of ovariectomized rats. An osteopenia model was used in 29 ovariectomized Wistar female rats with body weight of 250 g. Animals were divided in 6 experimental groups, the first group was sacrificed 30 days after the surgery and, the other groups, every 15 days on until the 105th day. The evaluation of installation process was made through anthropometrics and mechanical proprieties of femur (three-point bending test). Program GraphPad InStat 3 was used in order to compare the observed items. T-student test was used to check the body weight variation with p < 0,05 and tests ANOVA and Student-Newmam-Keuls were applied to the other items (p < 0,05). A significant increase in femur length and in maximum bending load were obtained. The found results in this research suggest that ovariectomy is a factor that does not cause significant mechanical and geometric alterations on cortical region of femurs in female rats during the analyzed period

The cortex can be viewed as a network of functional areas. A cortical area, composed ofneurons forming local connections, interacts with other areas via long distance connections.Each neuron receives multiple inputs and has to integrate the incoming signals. This integrativecapacity is the basis of the computational power of the brain. Our work concentrates onunderstanding the principles that govern the structure of the cortical network i.e. the allocationof neural resources as well as the anatomical segregation between processing steams. Usingretrograde tracer injections we extract two quantitative parameters: (i) the proportion ofSupragranular Labelled Neurons (SLN) identifies the feedforward (FF) or feedback (FB)operation between the source and target area; (ii) the Fraction of Labelled Neurons (FLN)identifies the magnitude of a connection pathway.We have made repeat injections in V1, V2, V4 to investigate the consistency of corticalpathways. This showed that (i) connection weights are consistent between animals; (ii) the listof areas projecting to each injection site is highly reproducible. We find that there are fixedFLN values for each pair of interconnected areas. The FLN values of all the afferent pathwaysto a given target span over a factor of 6 levels of log and although there is some overdispersiontheir variability is not larger than one single level of log meaning that there is a specificconnectivity profile for each area. Futermore the FLN follow a lognormal distribution. Inlognormals the mode is lower than the median and the mean i.e. the majority of pathways haveFLN weaker than the average FLN, meaning that strong projections are rare. If instead thedistribution of FLN was to follow a power law, then high FLN values would have been evenrarer. We found, a regularity in that the strongest input is invariably from within the injectedarea, second strongest are the inputs from areas sharing common borders with the target area.Sub-cortical inputs have a weak FLN, even when they are associated with an importantfunctional role such as the LGN → V1 pathway. We found that projection distance is inverselyrelated to the FLN value and an exponential distance rule operates that constrains short distanceprojections to high FLN and long distance projections to low FLN.We injected a total of 26 cortical areas homogenously distributed across the cortex. Thisrevealed 1232 projection pathways. Roughly 30% of pathways that we reveal have notpreviously been reported in the literature. Our ability to find new connections is due to theimproved tracing and brain segmentation techniques. We scan the whole brain at up to 80μmintervals to detect projection neurons, and this, as discussed in the text, is a major advantage toexisting studies. The weak long distance connections were shown to contract the characteristicpath-length of the graph (number of hops needed to go between any two areas).Our analysis of the graph showed that contrary to current belief the cortical inter-areal networkis dense (i.e. 58% of the connection that could exist do exist). At such a density, models basedon binary features such as small world cannot capture the specificity of the graph. Hence thecortex does not correspond small-world network, with sparse clustered graph possessingempowered by few critical projecitons that ensure short characteristic path-lengths. Furtheranalysis of pathway efficiency showed that the short distance connections of high magnitudeprovide large bandwidth for local connectivity and form a backbone of clustered functionallyrelated areas. This backbone is embedded in a sea of weak connections providing direct linksbetween cortical areas. We refer to this architecture as a tribal-network. We speculate that thesmall scale and high density that characterize the cortico-cortical network is facilitating theemergence of synchrony between cortical areas.