INTRODUÇÃO: O Transtorno do Espectro Autista tem incidência precoce e causas genéticas e ambientais multifatoriais. Gera disfunção no desenvolvimento da linguagem e da sociabilidade e um padrão de comportamento restrito e repetitivo. A formação e o desenvolvimento do sistema nervoso são afetados provocando impacto na arquitetura tecidual, desequilíbrio funcional entre os sistemas excitatórios e inibitórios, além de problemas na conectividade e consistência das redes neurais. O tratamento baseia-se em estimulação multiprofissional precoce e no uso de medicamentos que interferem na sintomatologia impactante do cotidiano. Os comportamentos restritos e repetitivos marcam o perfil funcional do paciente com Transtorno do Espectro Autista e podem interferir diretamente no desenvolvimento da linguagem e na sociabilidade. A mudança do espectro comportamental em razão do amadurecimento cognitivo não impede o desequilíbrio funcional nas atividades da vida diária. Existem indícios de que a função da flexibilidade cognitiva somada à capacidade para gerar novos conceitos e à velocidade de processamento das informações estão afetadas nesses pacientes e podem representar a base fisiopatológica dos Comportamentos Restritos e Repetitivos e suas repercussões no desenvolvimento desses indivíduos. O presente estudo visa avaliar os efeitos da facilitação da transmissão dos impulsos nervosos na função da flexibilidade cognitiva e no comportamento restrito e repetitivo dos pacientes com o diagnóstico de Transtorno do Espectro Autista de alto funcionamento através da Estimulação Magnética Transcraniana de pulsos pareados no Córtex Pré Frontal Ventro Lateral (CPFVL) direito em sua porção antero inferior. MÉTODOS: Trata-se de um estudo experimental com procedimento sistematizado e intervencional realizado com 11 pacientes de 9 a 17 anos, pareados por um perfil cognitivo dentro dos limites da normalidade, que foram submetidos ao protocolo de estimulação com a técnica de \"Theta Burst\" em CPFVL antero inferior à direita em 15 sessões divididas em três semanas. Os pacientes foram testados antes e depois do procedimento. A avaliação da flexibilidade cognitiva, da capacidade de formação de novos conceitos e do tempo cognitivo foram realizada através dos testes Wisconsin Sort Card Test (WSCT) e STROOP, aplicados aos pacientes. Os Comportamentos Restritos e Repetitivos foram avaliados pelos instrumentos Repetitive Behavior Scale - Revised (RBS-r) e Escala de Pensamentos Obsessivos e Comportamentos Compulsivos de Yale Brown (YBOCS) por meio de questionários aplicados aos pais e cuidadores desses pacientes. RESULTADOS: Encontraram-se evidências de mudanças no padrão de inflexibilidade cognitiva, representada pelo WSCT, nos itens Erros Perserverativos (p=0,028) e na Formação de Nível Conceitual (p=0,022). A alteração demonstrada pela Formação de Nível Conceitual pelo WSCT e pelo Total de Tempo para realizar o STROOP test (p=0,001) representam mudança na velocidade de processamento de informações. Constatou-se melhora do Comportamento Restrito e Repetitivo tanto pelo RBS-r (p=0,002) quanto no YBOCS Total de Compulsões (p=0,02). Todas as classes mostraram mudanças significativas: Ordem Inferior (p=0,05), Ordem superior (p=0,028) e Insistência na Mesmice (p=0,014). Houve correlação entre as curvas que representam as mudanças ocorridas no RBS-r e na flexibilidade cognitiva, representada pelo item Respostas Perseverativas (r=0,049). Por se tratar de um piloto, o presente estudo permite o aprofundamento das pesquisas que relacionam esta forma de intervenção descrita ao tratamento do Comportamento Restrito e Repetitivo em pacientes com Transtorno do Espectro Autista / INTRODUTION: The Autism Spectrum Disorder has an early incidence. It has genetic and environment multifactorial causes. Patients present language and social development disability and restricted and repetitive behavior. The maturation and development of the nervous system are damaged. Hence there is a disruption on the tissue architecture, functional imbalance between excitatory and inhibitory systems and connectivity and consistency of neural networks issues. The treatment is grounded on premature multiprofessional stimulation to improve cognition and the medication for mood instability, sleep problems, anxiety, hyperactivity and attention problems. The Restricted and Repetitive Behavior marks the functional profile of the Autism Spectrum Disorder and can hinder the communication and sociability development. The behavior changes with the cognitive maturation but does not prevent from the functional imbalance in daily activities. Evidence revel that the cognitive inflexibility added to the impairment to form new concepts and cognitive tempo can reflect the pathophysiology of these behaviors and its impact on development. The current study aims to valuate changes on cognitive flexibility and restricted and repetitive behavior after the facilitation of impulses through paired pulse Trascranial Magnetic Stimulation (EMT) on right anterior inferior Ventro Lateral Pre Frontal Cortex (VLPFC) in high function Autism Spectrum Disorder individuals. METHODS: It is an experimental study with systemic interventional procedure. Eleven subjects with nine to seventeen years old paired by normal cognitive profile were submitted to the protocol of Theta Burst stimulation on right anterior inferior VLPFC with fifteen sessions divided into three weeks. Subjects were tested before and after the procedure. They were tested for cognitive inflexibility, the ability to form new concepts and the cognitive tempo by the Wisconsin Sort Card (WSCT) test and STROOP test. The restricted and repetitive behaviors were evaluated by Restricted and Repetitive Behavior Scale revised (RBS-r) and Yale Brown Obsessive and Compulsive Scale applied for parents. RESULTS: Evidence of changes on cognitive inflexibility pattern were found items Perseverative Errors (p=0,028) and Conceptual level Response (p=0,022) of the WSCT test. Information processing speed changes were established by the Conceptual level Response of the WSCT and the Total Time to Finish the STROOP test (p=0,001). Improvement on restricted and repetitive behavior were found by changes on total score of RBS-r (p=0,002) and by YBOCS Total of Compulsions (p=0,02). There were changes on all aspects of the repetitive behaviors: Low-Order behavior (p=0,05), High-Order behavior (p=0,028) and Sameness behavior (0,014). Correlation were found between changing curves of cognitive inflexibility represented by Perseverative Response and total score of RBS-r (r=0,049). Since this is a pilot study, our data allows us to suggest that EMT-pp can be efficient for treating restricted and repetitive behavior on Autism Spectrum Disorder. Further controlled studies will be conducted to confirm this hypothesis

A tartaruga verde (Chelonia mydas) é uma tartaruga marinha que se alimenta e nidifica em locais da costa brasileira, ilhas e atóis. Esta espécie tem sido acometida pela fibropapilomatose (FP), doença considerada como uma das mais importantes ameaças à sua sobrevivência e caracterizada por tumores cutâneos benignos. A etiologia é multifatorial e complexa, possivelmente havendo relação entre Chelonid Fibropapilloma-associated Herpesvirus, fatores ambientais e genéticos. Foram estudadas 133 tartarugas verdes com e sem tumores provenientes de cinco áreas brasileiras: Florianópolis/SC, Ubatuba/SP, Vitória/ES, Almofala/CE e Fernando de Noronha/PE. Foram obtidos dados de biometria (comprimento curvilíneo da carapaça-CCC, massa corporal-MC e índice de condição corporal-ICC), tamanho (classificado por categorias A, B, C e D), quantidade e localização anatômica dos tumores, além da colheita de amostra sanguínea. Os objetivos foram (1) avaliar a atividade dos leucócitos por citometria de fluxo (fagocitose e burst oxidativo); (2) investigar a presença de sete congêneres de bifenilas policloradas (PCBs) e (3) estabelecer correlações entre PCBs e a atividade dos leucócitos. Houve diferenças no CCC entre tartarugas sem tumores de acordo com a área de estudo, na MC e no ICC entre animais acometidos ou não capturados em Ubatuba. Foram quantificados e classificados 1033 tumores de 50 tartarugas verdes, sendo que a quantidade por tartaruga variou de 1 a 61 e a maior parte foi classificada nas categorias de tamanho A (446 tumores) e B (520 tumores) e na região anterior (669 tumores). Foi observada diferença significativa nas concentrações do congênere PCB 138 entre tartarugas com e sem FP capturadas em Vitória. Animais acometidos tiveram maiores concentrações de todas as PCBs quantificadas, exceto as tartarugas verdes capturadas em Vitória no caso dos congêneres PCBs 118, 138, 153 e 180. PCB 138 foi quantificado em maior concentração nas amostras e PCB 180 em níveis menores. Não foram observadas correlações significativas entre concentrações de PCBs e atividade celular. A FP tem sido amplamente pesquisada e os estudos que visam estabelecer correlações entre poluentes químicos e saúde animal são extremamente importantes, pois podem contribuir para novas direções nos planos de ação para conservação. / The green sea turtle (Chelonia mydas) feeds and nests in Brazilian coast, islands and atolls. This species has been affected by fibropapillomatosis (FP), disease considered one of the most important threats to its survival and characterized by benign skin tumors. The etiology is multifactorial and complex, possibly having relation between Chelonid Fibropapilloma-associated Herpesvirus, environmental and genetic factors. Were studied 133 green sea turtles with and without FP from five Brazilian areas: Florianópolis/SC, Ubatuba/SP, Vitória/ES, Almofala/CE and Fernando de Noronha/PE. Were taken biometric data (curved carapace length-CCL, body mass-BM and body condition index-BCI), as well as size (classified by categories A, B, C and D), numbers and anatomical distribution of tumors, besides blood sample collection. The aims were (1) to assess leukocytes activity by flow cytometry (phagocytosis and oxidative burst); (2) to investigate the presence of seven congeners of polychlorinated biphenyls (PCBs) and (3) to establish correlations between PCBs and leukocytes activity. There were differences in CCL between turtles without FP according to the study area, in BM and BCI between affected and non-affected animals caught in Ubatuba. Were quantified and classified 1033 tumors of 50 green sea turtles, the number of tumors ranged from 1 to 61 and the most part of them was classified in categories of size A (446 tumors) and B (520 tumors) and in anterior region (669 tumors). Was observed difference in PCB 138 concentrations between green sea turtle with and without FP caught in Vitória. Affected animals had higher concentrations of all PCBs quantified, except green sea turtles caught in Vitória in the event of PCBs 118, 138, 153 and 180. Were quantified highest concentrations of PCB 138 and lowest of PCB 180. There were not significant correlations between PCBs concentration and leukocyte activity. The FP has been widely investigate and the studies that aimed to establish correlations between pollutants and animal health are very important, because can contribute for new directions in sea turtle conservation issues.

Bakgrund: Deep Brain Stimulation (DBS) i nucleus subthalamicus (STN) och i kaudala zona incerta (cZi) har visat positiva effekter på motoriska symtom (rörelseförmåga och balans) vid Parkinsons sjukdom. Efter DBS har dock negativa taleffekter noterats, inklusive nedsatt artikulatorisk precision och stabilitet. Nedsatt artikulatorisk precisionoch stabilitet påverkar klusilproduktionen mest vilket kan resultera i att klusilexplosionen uteblir eller att multipla explosioner uppstår. Mål: Att undersöka artikulatorisk precision och stabilitet vid klusilexplosionen efter DBS i STN respektive cZi. Metod: I studien deltog 19 patienter varav 9 deltagare stimulerats i STN och 10 deltagare i cZi. Talinspelningar gjordes innan operation och ett år efter operation med DBSstimulering på och av. Klusilerna i talmaterialet delades in i tre kategorier, de med en klusilexplosion, de med två eller flera klusilexplosioner samt de utan klusilexplosion. Hypotesprövning gjordes gällande fördelningen av klusiler med multipla explosioner samt klusiler utan explosion mellan och inom patientgrupperna samt inom respektive klusil. Resultat: Multipla explosioner ökade hos cZi-patienterna och minskade hos STNpatienterna. Klusiler som saknade explosion ökade hos STN-patienterna medan de minskade hos cZi-patienterna. För båda patientgrupperna ökade multipla explosioner och klusiler utan explosion då stimuleringen var på jämfört med av. Skillnaderna mellan och inom grupperna var inte signifikanta. Slutsats: Artikulatorisk precision och stabilitet påverkades efter DBS i form av bristande stabilitet, för kort slutningsrörelse och felaktig koordination av slutningsrörelsen. Detta resulterade i multipla explosioner och uteblivna explosioner med olika effekter för elektrodlokalisationerna. / Background: Deep Brain Stimulation (DBS) in Subthalamicus Nucleus (STN) and Caudala zona incerta (cZi) have shown positive effects on motor symptoms in Parkinson‘s disease. Negative effects on speech after DBS has been noted including reduced articulatory precision. Reduced articulatory precision and stability affects the production of stop consonants and as a result, loss of burst or multiple burst can occur. Aim: To investigate articulatory precision and stability regarding the burst in stop consonants after DBS of STN or cZi. Method: The study included 19 patients with 9 patients stimulated in STN and 10 patientsin cZi. Speech recordings were made before surgery and one year after with stimulation ON and OFF. The stop consonants were divided into three categories, those with one burst, those with two or more bursts (multiple burst) and those with loss of burst. Hypothesis testing was done on the categorization of the stop consonants in and between the groups and between the stop consonants types. Results: Multiple burst increased in the cZi group and decreased in the STN group. Stop consonants with loss of burst increased in the STN group but decreased in the cZi group. For both groups multiple burst and stop consonants with loss of burst increased with stimulation on. The differences between and within the groups were not significant. Conclusion: Articulatory precision and stability were affected by DBS with decreased stability, shortened occlusion and incorrect coordination. As a result multiple burst and loss of burst occurred in both groups. / Speech, voice and swallowing outcomes after deep brain stimulation of the zona incerta and the pedunculopontine nucleus in Parkinson’s disease: Comparsion with stimulation of the subthalamic nucleus

Ce travail a montré que l’apport de la technologie d’intégration 3D, permet de surmonter les limites imposées par la technologie monolithique sur les performances électriques (« coupling » et consommation) et sur l’implémentation physique (aire du pixel) des imageurs. Grâce à l’analyse approfondie sur la technologie d’intégration 3D, nous avons pu voir que les technologies d’intégration 3D les plus adaptées pour l’intégration des circuits dans le pixel sont : 3D wafer level et 3D construction séquentielle. La technologie choisie pour cette étude, est la technologie d'intégration 3D wafer level. Cela nous a permis de connecter 2 wafers par thermocompression et d’avoir une interconnexion par pixel entre wafers. L’étude de l’architecture CAN dans le pixel a montré qu’il existe deux limites dans le pixel : l’espace de construction et le couplage entre la partie analogique et numérique « digital coupling ». Son implémentation dans la technologie 3D autorise l’augmentation de 100% l’aire de construction et la réduction du « digital coupling » de 70%. Il a été implémenté un outil de calcul des éléments parasites des structures 3D. L’étude des imageurs rapides, a permis d’étendre l’utilisation de cette technologie. L’imageur rapide type « burst » a été étudié principalement. Cet imageur permet de dissocier la partie d’acquisition des images de la sortie. La limite principale, dans la technologie monolithique, est la taille des colonnes (pixels vers mémoires). Pour une haute cadence d’acquisition des images, il faut une grande consommation de courant. Son implémentation dans la technologie 3D a autorisé à mettre les mémoires au-dessous des pixels. Les études effectuées pour ce changement (réduction de la colonne à une interconnexion entre wafers), ont réduit la consommation totale de 90% et augmenté le temps d’acquisition des images de 184%, en comparaison à son pair monolithique. / This work has shown that the contribution of 3D integration technology allows to overcome the limitations imposed by monolithic technology on the electrical performances (coupling and consumption) and on the physical implementation (area of the pixel) of imagers. An in-depth analysis of the 3D integration technology has shown that the most suitable 3D integration technologies for the integration of the circuits at the pixel level are: 3D wafer level and 3D sequential construction. The technology chosen for this study is the 3D wafer level integration technology. This allows us to connect 2 wafers by thermocompression bonding and to have an interconnection or “bonding point” par pixel between wafers. The study of the architecture CAN at the pixel level showed that there are two limits in the pixel: the construction area and the coupling between the analog and digital part «digital coupling». Its implementation in 3D technology allows the construction area to be increased by 100% and the digital coupling reduced by 70%. It has been implemented a tool for computing the parasitic elements of 3D structures. The study of high speed imagers has allowed the use of this technology to be extended. The "burst" imager was mainly studied. This kind of imager’s architecture can dissociate the image acquisition from the output part. The main limit, in monolithic technology, is the size of the columns (pixels to memories). For a high rate of image acquisition, a high current consumption is required. Its implementation in 3D technology allowed to put the memories below the pixels. The studies carried out for this change (reduction of the column to an interconnection between wafers) reduced the total consumption by 90% and increased the acquisition time of the images by 184%, compared to its monolithic peer.

As infecções constituem a principal causa de morbidade e mortalidade em pacientes com lúpus eritematoso sistêmico (LES), representando 20-55% das mortes e sendo 80% delas causadas por bactérias. O LES é uma doença autoimune inflamatória crônica e a suscetibilidade às infecções está associada às próprias anormalidades imunológicas da doença, bem como a sua terapia, particularmente imunossupressora e citotóxica. Além disso, os polimorfismos genéticos dos Fc?R, Fc?RIIa e Fc?RIIIb, nos neutrófilos, têm sido associados com as disfunções imunes do LES.Os Fc?R são importantes mediadores das funções efetoras do neutrófilo e atuam em sinergismo com os CR. O polimorfismo dos genes FCGR2A e FCGR3B determina a expressão de variantes alélicas com diferenças funcionais, as quais podem influenciar as respostas biológicas e a suscetibilidade e o prognóstico das doenças infecciosas.O objetivo deste estudo foi avaliar a influência dos polimorfismos dos receptores Fc?RIIa (H/R131), Fc?RIIIb (HNA-1a, HNA-1b e HNA-1c) e CR3 (HNA-4a) no burst oxidativo de neutrófilos de pacientes com LES.Neutrófilos de pacientes com LES (n=36) e indivíduos saudáveis (n=36) foram purificados do sangue periférico (0.5x106/500µL) e estimulados com 30µg de IC, IC/soro humano normal (SHN), IC/SHN inativado ou PMA 10-7M. O burst oxidativo foi medido por quimioluminescência (QL) na presenca de luminol 10-4M ou lucigenina 10-4M. As frequências dos genótipos de Fc?RIIa, Fc?RIIIb e HNA-4a em pacientes com LES (n=157) e indivíduos saudáveis (n=147) foram determinadas por PCR com primers oligoespecíficos e gel de agarose 2%.Quanto aos polimorfismos genéticos, foi observado que o alelo positivo de HNA-4a contribui para a proteção e o alelo negativo para a suscetibilidade ao LES. Entre os pacientes com LES, as infecções foram mais frequentes quando os alelos R131 de FCGR2A, HNA-1b de FCGR3B e HNA-4a positivo do CR3 estavam presentes. Para o burst oxidativo com luminol, no grupo controle, as homozigoses H131, HNA-1b e HNA-4a negativo foram associadas à redução do burst oxidativo dos neutrófilos comparado às homozigoses para os respectivos alelos correspondentes. No LES, o burst oxidativo foi maior na homozigose R131 do grupo LES inativo comparado ao homozigoto H131 controle; menor na homozigose HNA-1b do grupo LES ativo comparado ao homozigoto HNA-1a do controle e, também, na heterozigose HNA-4a positivo/negativo o burst foi menor no grupo LES ativo comparado ao LES inativo. A ausência de diferenças entre os grupos com LES e controle, nos ensaios de burst oxidativo com lucigenina e com PMA, sugerem que a NADPH oxidase, responsável pela geração do burst oxidativo, não está comprometida nos neutrófilos dos pacientes com LES. Esses resultados têm implicações para a fisiopatologia do LES e, sobretudo, reforçam a hipótese de que os polimorfismos dos FCGR2A, FCGR3B e HNA-4a modulam o burst oxidativo de neutrófilos nos indivíduos saudáveis e no LES. Assim, o presente estudo contribui para o entendimento das anormalidades nas funções dos neutrófilos no LES. / Infections represent 20-55% of all deaths in patients with systemic lupus erythematosus (SLE). About 80% of them are caused by bacteria. SLE is an autoimmune disease in whichdisease-related and genetic factors and immunosuppressive and cytotoxic therapies all contributeto an increased susceptibility to infections.Recent data have provided evidence that genetic polymorphism of Fc?R is associated withimmune abnormalities and risk to development of SLE.Fc?R can mediate neutrophil effector functions and play a synergistic action with CR. Fc?RIIaand Fc?RIIIb display functionally relevant genetic polymorphisms, which allelic variants caninfluence the biological responses and the susceptibility to and course of infectious diseases.The aim of this study was to investigate the influence of the Fc?RIIa (H/R131), Fc?RIIIb (HNA- 1a, HNA-1b and HNA-1c) and CR3 (HNA-4a) polymorphisms on neutrophil oxidative burst of SLE patients. Neutrophils of SLE patients (n=36) and control individuals (n=36) were purified from peripheral blood (0.5x106/500µL) in Hanks 0.1% gelatin pH7.2 and were stimulated with 30µg of immune complexes (IC), IC/normal human serum (NHS), IC/heat-inactivated NHS (iNHS) or PMA 10-7M. The oxidative burst was measured by chemiluminescence (CL) in the presence of luminol 10-4M or lucigenin 10- 4 M. The reaction was monitored in a luminometer at 37ºC for 20min and the results were analyzed as the area under the curve of the CL profile. Genotype frequencies of Fc?RIIa, Fc?RIIIb and HNA-4a alleles for SLE patients (n=157) and for healthy subjects (n=147) were determined using PCR with sequence-specific primers and agarose gel 2%. It was observed that the HNA-4a positive allele contributes to the protection and the negative allele contributes to susceptibility to SLE. Among patients with SLE, infections were more frequent when the alleles R131 of FCGR2A, HNA-1b of FCGR3B and HNA-4a positive of CR3 were present. For the oxidative burst with luminol in the control group, the homozygous H131, HNA-1b and HNA-4a negative were associated with reduced oxidative burst of neutrophils compared to homozygous to their corresponding alleles. In SLE, the oxidative burst was higher in homozygous R131 inactive SLE group compared to the homozygous H131 control; it was also lower in homozygous HNA-1b active SLE group than homozygous HNA-1a control and either in the heterozygous HNA-4a positive/negative active SLE compared to inactive SLE group. The absence of differences between the SLE and control groups in the oxidative burst assays, using lucigenin and PMA, suggests that NADPH oxidase, responsible for generating the oxidative burst, is not impaired in neutrophils from patients with SLE. These results have implications for the pathophysiology of SLE and, above all, support the hypothesis that polymorphisms of FCGR2A, FCGR3B and HNA-4a modulate the oxidative burst of neutrophils in healthy and in SLE. Thus, this study contributes to the understanding of abnormalities in neutrophil functions in SLE.

INTRODUÇÃO: O Transtorno do Espectro Autista tem incidência precoce e causas genéticas e ambientais multifatoriais. Gera disfunção no desenvolvimento da linguagem e da sociabilidade e um padrão de comportamento restrito e repetitivo. A formação e o desenvolvimento do sistema nervoso são afetados provocando impacto na arquitetura tecidual, desequilíbrio funcional entre os sistemas excitatórios e inibitórios, além de problemas na conectividade e consistência das redes neurais. O tratamento baseia-se em estimulação multiprofissional precoce e no uso de medicamentos que interferem na sintomatologia impactante do cotidiano. Os comportamentos restritos e repetitivos marcam o perfil funcional do paciente com Transtorno do Espectro Autista e podem interferir diretamente no desenvolvimento da linguagem e na sociabilidade. A mudança do espectro comportamental em razão do amadurecimento cognitivo não impede o desequilíbrio funcional nas atividades da vida diária. Existem indícios de que a função da flexibilidade cognitiva somada à capacidade para gerar novos conceitos e à velocidade de processamento das informações estão afetadas nesses pacientes e podem representar a base fisiopatológica dos Comportamentos Restritos e Repetitivos e suas repercussões no desenvolvimento desses indivíduos. O presente estudo visa avaliar os efeitos da facilitação da transmissão dos impulsos nervosos na função da flexibilidade cognitiva e no comportamento restrito e repetitivo dos pacientes com o diagnóstico de Transtorno do Espectro Autista de alto funcionamento através da Estimulação Magnética Transcraniana de pulsos pareados no Córtex Pré Frontal Ventro Lateral (CPFVL) direito em sua porção antero inferior. MÉTODOS: Trata-se de um estudo experimental com procedimento sistematizado e intervencional realizado com 11 pacientes de 9 a 17 anos, pareados por um perfil cognitivo dentro dos limites da normalidade, que foram submetidos ao protocolo de estimulação com a técnica de \"Theta Burst\" em CPFVL antero inferior à direita em 15 sessões divididas em três semanas. Os pacientes foram testados antes e depois do procedimento. A avaliação da flexibilidade cognitiva, da capacidade de formação de novos conceitos e do tempo cognitivo foram realizada através dos testes Wisconsin Sort Card Test (WSCT) e STROOP, aplicados aos pacientes. Os Comportamentos Restritos e Repetitivos foram avaliados pelos instrumentos Repetitive Behavior Scale - Revised (RBS-r) e Escala de Pensamentos Obsessivos e Comportamentos Compulsivos de Yale Brown (YBOCS) por meio de questionários aplicados aos pais e cuidadores desses pacientes. RESULTADOS: Encontraram-se evidências de mudanças no padrão de inflexibilidade cognitiva, representada pelo WSCT, nos itens Erros Perserverativos (p=0,028) e na Formação de Nível Conceitual (p=0,022). A alteração demonstrada pela Formação de Nível Conceitual pelo WSCT e pelo Total de Tempo para realizar o STROOP test (p=0,001) representam mudança na velocidade de processamento de informações. Constatou-se melhora do Comportamento Restrito e Repetitivo tanto pelo RBS-r (p=0,002) quanto no YBOCS Total de Compulsões (p=0,02). Todas as classes mostraram mudanças significativas: Ordem Inferior (p=0,05), Ordem superior (p=0,028) e Insistência na Mesmice (p=0,014). Houve correlação entre as curvas que representam as mudanças ocorridas no RBS-r e na flexibilidade cognitiva, representada pelo item Respostas Perseverativas (r=0,049). Por se tratar de um piloto, o presente estudo permite o aprofundamento das pesquisas que relacionam esta forma de intervenção descrita ao tratamento do Comportamento Restrito e Repetitivo em pacientes com Transtorno do Espectro Autista / INTRODUTION: The Autism Spectrum Disorder has an early incidence. It has genetic and environment multifactorial causes. Patients present language and social development disability and restricted and repetitive behavior. The maturation and development of the nervous system are damaged. Hence there is a disruption on the tissue architecture, functional imbalance between excitatory and inhibitory systems and connectivity and consistency of neural networks issues. The treatment is grounded on premature multiprofessional stimulation to improve cognition and the medication for mood instability, sleep problems, anxiety, hyperactivity and attention problems. The Restricted and Repetitive Behavior marks the functional profile of the Autism Spectrum Disorder and can hinder the communication and sociability development. The behavior changes with the cognitive maturation but does not prevent from the functional imbalance in daily activities. Evidence revel that the cognitive inflexibility added to the impairment to form new concepts and cognitive tempo can reflect the pathophysiology of these behaviors and its impact on development. The current study aims to valuate changes on cognitive flexibility and restricted and repetitive behavior after the facilitation of impulses through paired pulse Trascranial Magnetic Stimulation (EMT) on right anterior inferior Ventro Lateral Pre Frontal Cortex (VLPFC) in high function Autism Spectrum Disorder individuals. METHODS: It is an experimental study with systemic interventional procedure. Eleven subjects with nine to seventeen years old paired by normal cognitive profile were submitted to the protocol of Theta Burst stimulation on right anterior inferior VLPFC with fifteen sessions divided into three weeks. Subjects were tested before and after the procedure. They were tested for cognitive inflexibility, the ability to form new concepts and the cognitive tempo by the Wisconsin Sort Card (WSCT) test and STROOP test. The restricted and repetitive behaviors were evaluated by Restricted and Repetitive Behavior Scale revised (RBS-r) and Yale Brown Obsessive and Compulsive Scale applied for parents. RESULTS: Evidence of changes on cognitive inflexibility pattern were found items Perseverative Errors (p=0,028) and Conceptual level Response (p=0,022) of the WSCT test. Information processing speed changes were established by the Conceptual level Response of the WSCT and the Total Time to Finish the STROOP test (p=0,001). Improvement on restricted and repetitive behavior were found by changes on total score of RBS-r (p=0,002) and by YBOCS Total of Compulsions (p=0,02). There were changes on all aspects of the repetitive behaviors: Low-Order behavior (p=0,05), High-Order behavior (p=0,028) and Sameness behavior (0,014). Correlation were found between changing curves of cognitive inflexibility represented by Perseverative Response and total score of RBS-r (r=0,049). Since this is a pilot study, our data allows us to suggest that EMT-pp can be efficient for treating restricted and repetitive behavior on Autism Spectrum Disorder. Further controlled studies will be conducted to confirm this hypothesis

A tartaruga verde (Chelonia mydas) é uma tartaruga marinha que se alimenta e nidifica em locais da costa brasileira, ilhas e atóis. Esta espécie tem sido acometida pela fibropapilomatose (FP), doença considerada como uma das mais importantes ameaças à sua sobrevivência e caracterizada por tumores cutâneos benignos. A etiologia é multifatorial e complexa, possivelmente havendo relação entre Chelonid Fibropapilloma-associated Herpesvirus, fatores ambientais e genéticos. Foram estudadas 133 tartarugas verdes com e sem tumores provenientes de cinco áreas brasileiras: Florianópolis/SC, Ubatuba/SP, Vitória/ES, Almofala/CE e Fernando de Noronha/PE. Foram obtidos dados de biometria (comprimento curvilíneo da carapaça-CCC, massa corporal-MC e índice de condição corporal-ICC), tamanho (classificado por categorias A, B, C e D), quantidade e localização anatômica dos tumores, além da colheita de amostra sanguínea. Os objetivos foram (1) avaliar a atividade dos leucócitos por citometria de fluxo (fagocitose e burst oxidativo); (2) investigar a presença de sete congêneres de bifenilas policloradas (PCBs) e (3) estabelecer correlações entre PCBs e a atividade dos leucócitos. Houve diferenças no CCC entre tartarugas sem tumores de acordo com a área de estudo, na MC e no ICC entre animais acometidos ou não capturados em Ubatuba. Foram quantificados e classificados 1033 tumores de 50 tartarugas verdes, sendo que a quantidade por tartaruga variou de 1 a 61 e a maior parte foi classificada nas categorias de tamanho A (446 tumores) e B (520 tumores) e na região anterior (669 tumores). Foi observada diferença significativa nas concentrações do congênere PCB 138 entre tartarugas com e sem FP capturadas em Vitória. Animais acometidos tiveram maiores concentrações de todas as PCBs quantificadas, exceto as tartarugas verdes capturadas em Vitória no caso dos congêneres PCBs 118, 138, 153 e 180. PCB 138 foi quantificado em maior concentração nas amostras e PCB 180 em níveis menores. Não foram observadas correlações significativas entre concentrações de PCBs e atividade celular. A FP tem sido amplamente pesquisada e os estudos que visam estabelecer correlações entre poluentes químicos e saúde animal são extremamente importantes, pois podem contribuir para novas direções nos planos de ação para conservação. / The green sea turtle (Chelonia mydas) feeds and nests in Brazilian coast, islands and atolls. This species has been affected by fibropapillomatosis (FP), disease considered one of the most important threats to its survival and characterized by benign skin tumors. The etiology is multifactorial and complex, possibly having relation between Chelonid Fibropapilloma-associated Herpesvirus, environmental and genetic factors. Were studied 133 green sea turtles with and without FP from five Brazilian areas: Florianópolis/SC, Ubatuba/SP, Vitória/ES, Almofala/CE and Fernando de Noronha/PE. Were taken biometric data (curved carapace length-CCL, body mass-BM and body condition index-BCI), as well as size (classified by categories A, B, C and D), numbers and anatomical distribution of tumors, besides blood sample collection. The aims were (1) to assess leukocytes activity by flow cytometry (phagocytosis and oxidative burst); (2) to investigate the presence of seven congeners of polychlorinated biphenyls (PCBs) and (3) to establish correlations between PCBs and leukocytes activity. There were differences in CCL between turtles without FP according to the study area, in BM and BCI between affected and non-affected animals caught in Ubatuba. Were quantified and classified 1033 tumors of 50 green sea turtles, the number of tumors ranged from 1 to 61 and the most part of them was classified in categories of size A (446 tumors) and B (520 tumors) and in anterior region (669 tumors). Was observed difference in PCB 138 concentrations between green sea turtle with and without FP caught in Vitória. Affected animals had higher concentrations of all PCBs quantified, except green sea turtles caught in Vitória in the event of PCBs 118, 138, 153 and 180. Were quantified highest concentrations of PCB 138 and lowest of PCB 180. There were not significant correlations between PCBs concentration and leukocyte activity. The FP has been widely investigate and the studies that aimed to establish correlations between pollutants and animal health are very important, because can contribute for new directions in sea turtle conservation issues.

Les Encéphalopathies Épileptiques Précoces sont des pathologies rares et sévères caractérisées par des crises fréquentes commençant dans les trois premiers mois de vie accompagnées d’un EEG intercritique altéré et un pronostic très défavorable. Au cours de la caractérisation génétique d’une cohorte de 402 patients, nous avons mis en évidence une délétion de 19,9 kb localisée en Xp11.23 chez un garçon et 34 mutations de novo du gène KCNQ2. La première partie de mon projet a consisté en l’étude de la pathogénicité de la délétion Xp11.23, qui implique trois gènes dont WDR45. Les mutations de WRD45 ont été décrites dans une dégénérescence neuronale avec accumulation de fer et presque exclusivement chez des patients de sexe féminin. Le diagnostic initial, chez ce patient, montre une IRM normale avec un phénotype d'EEP et l'accumulation de fer a été détectée à partir de 5 ans. Ce travail m’a permis de décrire le premier patient atteint d’EEP porteur d’une délétion de WDR45. La deuxième partie de mon projet a concerné le gène KCNQ2. Nos résultats ont montré que les mutations sont impliquées dans deux mécanismes physiopathologiques, une délocalisation subcellulaire et un gain de fonction. Ces résultats ouvrent de nouvelles perspectives en terme de compréhension de la pathologie et de thérapies qui peuvent être proposées. Une dernière partie de ce projet a consisté en l’élaboration de nouveaux modèles in vitro, j’ai mis au point des lignées stables exprimant KCNQ2 qui permettront le criblage de molécules thérapeutiques à haut-débit, ainsi que des progéniteurs neuronaux différenciés à partir de cellules iPS issues de la reprogrammation de fibroblastes de patients. / Early onset epileptic encephalopathies are rare and severe disorders, characterized by frequent motor seizures occurring before three months of age associated with an altered interictal EEG pattern. The prognosis is poor. During the course of the genetic characterization of a cohort of 402 EOEE patients, we identified a de novo deletion located at Xp11.23 in a male patient and 34 KCNQ2 de novo mutations. The first part of my project consisted in the study of the pathogenicity of the Xp11.23 deletion that encompasses three genes including WDR45. Mutations in the WDR45 gene been have recently identified in patients suffering from neurodegeneration with brain iron accumulation. WDR45 mutations have been almost exclusively found in females. Our patient with the Xp11.23 deletion presented a normal MRI and the EOEE phenotype was predominant. Iron accumulation began only at 5 years. My work reveals that deletions of WDR45 are viable in males and can be diagnosed as EOEE. The second part of my project was aimed at the functional study of two KCNQ2 gene mutations. During this work, my results showed that those mutations were involved in new pathological mechanisms, namely a mislocalization or gain of function. Those results provide new perspectives in term of disease knowledge and therapy. The last part of my project consisted in the development of two new in vitro models for the study of KCNQ2 mutations: stable cell lines expressing the Kv7.2 channel for high-throughput screening of drugs and the production of neurons from induced pluripotent stem cells arising from reprogrammed patient fibroblasts.

O presente trabalho tem o objetivo principal de compreender os possíveis efeitos da radiação energética de um evento de Gamma-Ray Burst (GRB) sobre o meio interestelar no entorno de seu local de geração e em planetas possivelmente iluminados. Gamma-Ray Bursts foram detectados pela primeira vez nos anos 60 e rapidamente atraíram a atenção da comunidade astrofísica, uma vez que as energias emitidas apenas em poderiam alcançar 1054erg, o equivalente a massa de repouso do Sol. Não se conhecia nenhum mecanismo tão eficiente para extrair energia gravitacional para produzir tal evento. Mais tarde, a possibilidade da emissão ser colimada abaixou a energia em para 5x1050erg, mas o mecanismo central de geração ainda não foi completamente desvendado, havendo muito espaço para alternativas exóticas. Estudamos os efeitos de um GRB sobre o meio interestelar, em uma tentativa de distinguir os remanescentes do GRB do gerado por múltiplas supernovas. Usamos argumentos energéticos e sobre a possibilidade de alterações químicas e isotópicas devido a reações fotonucleares. Também trabalhamos com as implicações biológicas da iluminação de planetas por um GRB, concluindo que os efeitos de tais eventos podem afetar seriamente a biosfera de um planeta mesmo a distâncias de ~10kpc. / The present work has the main goal of understanding the possible effects of the hard gamma radiation produced during a Gamma-Ray Burst (GRB) event both on the interstellar medium surrounding the source of the burst and on planets possibly illuminated. Gamma-Ray Bursts were first detected on the 60s and quickly have attracted the attention of the astrophysical community, since the energies emitted just in could reach 1054erg, the rest mass of the Sun. No mechanism was known to be so efficient in extracting gravitational energy to produce such emission. Later on, the possibility of the emission being collimated has lowered the energy of the to 5x1051erg, but the central engine has not yet been completely understood, and there is still ample room for exotic alternatives. We have studied the effects of GRB on the ISM, in an attempt to distinguish the candidates of GRB remnants from those generated by multiple supernovae. We have used both energetic arguments and the possibility of chemical alterations due to photonuclear reactions. We have also worked on the biological implications of the illumination of planets by a GRB, concluding that the effects of such event could seriously harm the biosphere of a planet even at distances of ~10kpc.

Ionenkanäle leiten Ionenströme über neuronale Membranen, wodurch Aktionspotentiale erzeugt und weitergeleitet werden. Sie spielen eine zentrale Rolle bei der Regulierung der Erregbarkeit und des Aktivierungsverhaltens von Neuronen. KCNQs sind eine wichtige Familie von spannungsgesteuerten Kaliumkanälen; ihre Dysfunktion kann zu verschiedenen neurologischen Krankheiten führen, einschließlich Erkrankung an Epilepsie und Taubheit. Es wurde gezeigt, dass KCNQ2 und KCNQ3 den M-Strom verantwortlich sind. Letzterer ist für die Regulierung des repetitiven Feuerns von Pyramidenzellen entscheidend. Im Gegensatz zu KCNQ2, ist die funktionelle Bedeutung von KCNQ3 noch nicht aufgeklärt. In dieser Arbeit zeigen wir mittels extrazellulärer Elektrophysiologie in vivo, dass bei konstitutiven Kcnq3 Knockoutmäusen die hippokampalen Pyramidenzellen vermehrt burstartig feuern. Außerdem weisen diese Tiere eine verminderte Spike-Frequenz-Anpassung auf und die Wahrscheinlichkeit des Burst-Feuerns während zwei verschiedener Oszillationen – Theta gegen Nicht-Theta – kann nicht mehr unterscheiden werden. Des Weiteren zeigen Kcnq3-Knockout- Pyramidenzellen während der Theta-Oszillation weder eine dominante Phasenpräferenz, noch eine Koordination ihrer Burst-Feuerung. Die Thetawellen Phasenpräzision tritt weiterhin bei dem vorübergehend verstärkten Feuern auf. Das räumliche selektive Feuern von mutmaßlichen Ortszellen blieb auch bei den Knockout-Mäusen erhalten, aber es ist hauptsächlich vom Burst- Feuern abhängig. Diese Studie zeigt, dass der KCNQ3-Ionenkanal eine wichtige Rolle bei der Regulierung der neuronalen Erregbarkeit und der Informationsverarbeitung spielt, und gibt damit Einblicke in die Bedeutsamkeit der KCNQ3-Ionenkanäle bezüglich der neurologischen Störungen. / Ion channels conduct ion flows across neuronal membrane whereby action potential is generated and propagated. They play a central role in regulating the excitability and firing behavior of a neuron. Among them, the KCNQs present a prominent family of voltage-gated potassium channels. Dysfunction of KCNQ2–5 channels can lead to varied neurological diseases including early onset epilepsy and deafness. In cortex and hippocampus, KCNQ2 and KCNQ3 have been demonstrated to underlie the non-inactivating M-current critical for controlling the repetitive firing of pyramidal cells. However, the functional significance of KCNQ3, unlike that of KCNQ2, remains elusive. Here, by applying in vivo extracellular electrophysiology in Kcnq3 constitutive knockout mice and the wild-type littermates, we find that hippocampal pyramidal cells lacking KCNQ3 exhibit increased burst firing. Moreover, the spike frequency adaptation of their bursts is diminished, and the burst propensity during two different field oscillations – theta versus non-theta – becomes indistinguishable. During theta oscillations, Kcnq3 knockout pyramidal cells no longer display unimodal phase preference and do not coordinate their burst firing. But phase advancement along successive theta cycles continues to occur at times of transiently intensified firing. The selective firing of place cells is largely preserved in the knockout while mainly relying on bursts. These results suggest that KCNQ3 channels indeed play a significant and specific role in regulating the neurons’ excitability and information processing, thus providing crucial mechanistic insights into the relevance of the KCNQ3 channels in neurological disorders.