46,XX doença do desenvolvimento sexual testicular : a propósito de um caso clínico

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Síndrome de bronquiolite obliterante após o transplante pulmonar : factores de risco

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Doença de Hirschsprung : abordagem sumária de um caso clínico

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014

Relação do polimorfismo BDNF val66met e níveis periféricos de BDNF com a doença de Parkinson e sua sintomatologia

Neurodegenerative diseases are frequently studied due to the increasing number of cases associated with the populational ageing and to the impact on the conditions on the quality of life. Parkinson’s disease (DP) is the second most frequent neurodegenerative disease. Despite the fact that its etiology is not completely understood, it is known that DP is caused by environmental and genetic factors. Thus, the investigation of etiologic factors and mechanisms responsible for the changes that lead to DP may help early diagnostic and prevention. A possible association between DP and the common polymorphism of Brain Derived Neurotrophic Factor (BDNF) G196A (Val66Met) has been suggested by different studies with contrasting results. For this reason, the aim of this study is to investigate if the BDNF Val66Met polymorphism is related to susceptibility to DP in a cohort of Brazilian patients. Additionaly, we verify if the presence of the polymorphism implies in alterations in the BDNF whole blood concentrations, as well as variations in symptomatology. The sample comprised Brazilian patients accompanied by the neurology service of the Onofre Lopes University Hospital (HUOL) and healthy controls (CTRL). The motor aspects of DP were evaluated by Hoehn e Yahr Scale (HY), Unified Parkinson’s Disease Rating Scale (UPDRS) and Schwab & England Scale (SE). For the evaluation of non-motor symptoms were used the following instruments: Frontal Assessment Battery (BAF), Mini-Mental State Examination (MEEM), Beck Depression Inventory (IDB) and the Beck Anxiety Inventory (IAB). Blood samples were collected for BDNF Val66Met polymorphism genotyping and BDNF whole blood measurement. As expected, DP patients performed worse in motor, cognitive and emotional battery of questionnaires. Alleles distribution between DP and CTRL was not significantly different, but the A/G genotype was significantly associated with a protector factor for DP. In contrast, the G/G genotype was significantly associated with depression and anxiety development in DP patients. However, BDNF concentrations were not different between genotypes or groups. This is the first study of genetic association of this polymorphism with DP in Brazilian subjects and the first one that associate A/G genotype with protection against DP.

Variabilidade genética de Leishmania spp. circulantes entre seres humanos e cães e infecção de flebotomíneos em áreas endêmicas para as leishmanioses no Rio Grande do Norte

Leishmania infantum is the main etiologic agent of visceral leishmaniasis in the New World. The pattern of distribution of leishmaniasis has changed substantially and has presented an emerging profile within the periphery of the Large Urban Centers. Leishmania infection can compromise skin, mucosa and viscera. Only 10% of the individuals infected develop the disease and 90% of human infection is asymptomatic. The main factors involved in the development of the disease are the host immune response, the vector’s species and the parasite’s genetic content. The sequencing of Leishmania isolated seeks to increase the understanding of the symptoms of individuals. The aim of this study was to evaluate the genetic diversity of circulating Leishmania strains among humans, and symptomatic and asymptomatic, and dogs from endemic areas of Rio Grande do Norte State and analyze sandflies from endemic areas for cutaneous and visceral disease. The genetic variability was evaluated by the use of markers hsp70 , ITS1 and a whole genome sequencing was also carried out. The amplified hsp70 and ITS1 of samples were analyzed and assembled using a Phred / Phrap package. The dendograms were constructed using the same methodology, but adding 500 bootstraps, followed by inferences on the relationships between Leishmania variants. The sequences of the 20 Brazilian isolates were mapped to the reference genome L. infantum JPCM5, using the Bowtie2 program and the identification of 36 contigs. The information of the valid SNPs were used in the PCA. SNPs were visualized by Geneious 7.1 and IGV. The genome annotations were transferred to their respective chromosomes and displayed on Geneious. The matching sequences of all chromosomes were aligned using Mauve. The phylogenetic trees were calculated according to maximum likelihood and JTT models. Sandflies were analyzed by PCR for the identification of Leishmania infection, a blood meal source and GAPDH sand fly. As a result, hsp70 and ITS1 were not capable of identifying genetic variability among human isolates from symptomatic and asymptomatic, and dogs. The complete sequencing of the 20 Brazilian isolates revealed a strong similarity between the circulating Leishmania strains in Rio Grande do Norte. The isolates collected in the city of Natal from humans and canines remained grouped in all analyzes, suggesting that there is genotypic and geographic proximity among the isolates. The isolated samples in the 1990s had a higher genotypic diversity when compared to freshly isolated samples. All isolates presented 36 chromosomes with variable ploidy among them, no correlation was found between the number of amastina genes copies, gp63, A2 and SSG with such clinic forms. In general, we did not find correlation between symptomatic and asymptomatic clinical forms and the gene content of the Brazilian isolates of Leishmania. 34,28% of the sandflies collected in the upper west region were L. longipalpis and the main sources of blood meal were humans, dogs and chickens.

Variabilidade genética de Leishmania spp. circulantes entre seres humanos e cães e infecção de flebotomíneos em áreas endêmicas para as leishmanioses no Rio Grande do Norte

Leishmania infantum is the main etiologic agent of visceral leishmaniasis in the New World. The pattern of distribution of leishmaniasis has changed substantially and has presented an emerging profile within the periphery of the Large Urban Centers. Leishmania infection can compromise skin, mucosa and viscera. Only 10% of the individuals infected develop the disease and 90% of human infection is asymptomatic. The main factors involved in the development of the disease are the host immune response, the vector’s species and the parasite’s genetic content. The sequencing of Leishmania isolated seeks to increase the understanding of the symptoms of individuals. The aim of this study was to evaluate the genetic diversity of circulating Leishmania strains among humans, and symptomatic and asymptomatic, and dogs from endemic areas of Rio Grande do Norte State and analyze sandflies from endemic areas for cutaneous and visceral disease. The genetic variability was evaluated by the use of markers hsp70 , ITS1 and a whole genome sequencing was also carried out. The amplified hsp70 and ITS1 of samples were analyzed and assembled using a Phred / Phrap package. The dendograms were constructed using the same methodology, but adding 500 bootstraps, followed by inferences on the relationships between Leishmania variants. The sequences of the 20 Brazilian isolates were mapped to the reference genome L. infantum JPCM5, using the Bowtie2 program and the identification of 36 contigs. The information of the valid SNPs were used in the PCA. SNPs were visualized by Geneious 7.1 and IGV. The genome annotations were transferred to their respective chromosomes and displayed on Geneious. The matching sequences of all chromosomes were aligned using Mauve. The phylogenetic trees were calculated according to maximum likelihood and JTT models. Sandflies were analyzed by PCR for the identification of Leishmania infection, a blood meal source and GAPDH sand fly. As a result, hsp70 and ITS1 were not capable of identifying genetic variability among human isolates from symptomatic and asymptomatic, and dogs. The complete sequencing of the 20 Brazilian isolates revealed a strong similarity between the circulating Leishmania strains in Rio Grande do Norte. The isolates collected in the city of Natal from humans and canines remained grouped in all analyzes, suggesting that there is genotypic and geographic proximity among the isolates. The isolated samples in the 1990s had a higher genotypic diversity when compared to freshly isolated samples. All isolates presented 36 chromosomes with variable ploidy among them, no correlation was found between the number of amastina genes copies, gp63, A2 and SSG with such clinic forms. In general, we did not find correlation between symptomatic and asymptomatic clinical forms and the gene content of the Brazilian isolates of Leishmania. 34,28% of the sandflies collected in the upper west region were L. longipalpis and the main sources of blood meal were humans, dogs and chickens.

Avaliação demográfica, clinico-laboratorial e genética de indivíduos com lupus eritematoso sistêmico e artrite reumatóide residentes em região tropical

The aetiology of autoimmunes disease is multifactorial and involves interactions among environmental, hormonal and genetic factors. Many different genes may contribute to autoimmunes disease susceptibility. The major histocompatibility complex (MHC) genes have been extensively studied, however many non-polymorphic MHC genes have also been reported to contribute to autoimmune diseases susceptibility. The aim of the present study was to evaluate the influence of SLC11A1 gene in systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA). Ninety-six patients with SLE, 37 with RA and 202 controls enrolled in this case-control study, were evaluated with regard to demographic, genetic, laboratorial and clinical data. SLE mainly affects females in the ratio of 18 women for each man, 88,3% of the patients aged from 15 to 45 years old and it occurs with similar frequency in whites and mulattos. The rate of RA between women and men was 11:1, with 77,1% of the cases occurring from 31 to 60 years. The genetic analysis of the point mutation -236 of the SLC11A1 gene by SSCP did not show significant differences between alleles/genotypes in patients with SLE or RA when compared to controls. The most frequent clinical manifestations in patients with SLE were cutaneous (87%) and joint (84.9%). In patients with RA, the most frequent out-joint clinical manifestation were rheumatoid nodules (13,5%). Antinuclear antibodies were present in 100% of the patients with SLE. There was no significant relation between activity of disease and presence of rheumatoid factor in patients with RA, however 55,6% of patients with active disease presented positive rheumatoid factor. Significant association between alleles/genotypes of point mutation -236 and clinical manifestations was not found

No tempo de Asclépio : a suposta engenharia genética de Platão

A descrição do papel da medicina no Livro III da República pode facilmente deixar o leitor constrangido. O trecho defende que a medicina deveria se espelhar nas práticas do tempo de Asclépio. Neste tempo, a medicina tinha um papel político de não prolongar a vida dos cidadãos tomados pela doença, nem deixá-los procriar. Frente a estas declarações, surgiram várias interpretações que vão desde o totalitarismo de Popper até a ironia de Strauss. Para uma melhor compreensão do texto, seria necessária a interlocução com a medicina hipocrática e com as práticas médicas do tempo dos séculos V e IV aEN. Contudo, a passagem em questão não se reduz ao contexto histórico, mas apresenta uma crítica válida ainda na atualidade. _______________________________________________________________________________ ABSTRACT

Identification of regulatory polymorphisms associated with breast cancer risk

Dissertação de Mestrado, Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, 2014

Melhoramento genético florestal e o desafio imposto pela doença da murchidão do pinheiro

Mestrado em Engenharia Florestal e dos Recursos Naturais - Instituto Superior de Agronomia - UL

Pesquisas genéticas, prognósticos morais e discriminação genética : um estudo de caso sobre traço falciforme

O debate sobre ética em pesquisa pode ser aplicado tanto ao âmbito da metodologia científica como a outras áreas do saber, como é o caso dos esportes. No campo da saúde esportiva brasileira, têm sido comuns pesquisas que fazem testes genéticos para identificar atletas com traço falciforme. Apesar da persistência de confederações esportivas brasileiras em discriminar atletas com essa característica hereditária, o traço falciforme não é uma doença. Este artigo relata o caso de uma atleta de futebol vítima de discriminação genética: identificada com o traço falciforme, ela foi considerada inapta a participar de um campeonato pela Confederação Brasileira de Futebol. O artigo analisa as repercussões da pesquisa genética para identificação do traço falciforme na ausência de cuidados éticos voltados à preservação dos direitos de quem se submete aos testes. Além disso, mostra a situação de vulnerabilidade à qual estão expostas pessoas envolvidas em pesquisas que fazem testes genéticos sem cuidados éticos ou mesmo justificativas razoáveis e cujos resultados são interpretados sob a racionalidade do determinismo biológico e do reducionismo genético. As confederações esportivas brasileiras interessadas em identificar atletas com o traço falciforme deveriam submeter esse objetivo de estudo à avaliação de Comitês de Ética em Pesquisa, pois esse é um procedimento com potencial de acarretar prejuízos aos atletas. O teste genético não pode ser considerado um ato de assistência em saúde, visto que não há doença a ser tratada. _______________________________________________________________________________ ABSTRACT

Diversidade genética estimada através de marcadores ISSR de Colletotrichum graminicola no Brasil.

A antracnose do milho causada pelo fungo Colletotrichum graminicola é uma das principais doenças da cultura no Brasil e no mundo, atacando praticamente todas as partes da planta. Neste trabalho, foi avaliada a variabilidade genética de 95 isolados monospóricos de C. graminicola provenientes dos estados de Goiás, Minas Gerais, Santa Catarina, São Paulo, Paraná e Rio Grande do Sul. Foram avaliados 15 primers ISSR, sendo selecionados nove que resultaram em um maior polimorfismo. Os fragmentos de DNA gerados pelas análises de ISSR foram avaliados mediante inspeção visual dos géis. Bandas de mesmo peso molecular, em indivíduos diferentes, foram consideradas idênticas e designadas em função da ausência (0) e presença (1) no gel. Baseado na matriz, foi gerado um dendrograma pelo método UPGMA, utilizando as 66 bandas amplificadas pelos nove primers ISSR. Ao analisar o dendrograma, foi traçada uma linha divisória no valor da distância de dissimilaridade de 0,3, dividindo os isolados em sete grupos. Baseado nos resultados, foi possível concluir que a variabilidade genética entre os isolados de C. graminicola é alta, sendo os marcadores ISSR eficazes na determinação de sua variabilidade. Os isolados utilizados no presente trabalho não apresentam estruturação geográfica.

Identificação de fonte de resistência à ferrugem polissora em milho para desenvolvimento de população de mapeamento e estudo de herança genética.

2016

Determinación de la incidencia de parásitos internos en los bovinos del Complejo uno y dos de la Empresa Genética Agenor Gómez, Boaco, Nicaragua

El presente trabajo se realizó en el periodo comprendido entre Noviembre y Diciembre de 1989, con el objetivo de investigar la infestación de parásitos gastrointestinales y pulmonares de mayor importancia económica en los bovinos del complejo 1 y 2 de la empresa genética Agenor Gómez. Boaco, Nicaragua. Se muestrearon 242 animales, entre bovinos de la raza Aberdeen Angus y Brahman, distribuidos en las categorías siguientes: C1 (0-1 año), C2 (1-3 años) y C3 (>3 años). Las muestras fueron extraídas directamente del recto animal, luego refrigeradas a 5°C y fueron analizadas en laboratorio de parasitología del MIDINRA, localizado en municipio de Camoapa V región. La cantidad de huevos y larvas se registró para cada género empleando las técnicas de sedimentación en agua, flotación con solución salina saturada y el método de larvoscopia e identificación con fotografías. Al realizar el ANDEVA por género de parásitos reportados en el análisis coprológico para la variable N° de huevos y larvas/gr de heces, se encontró que existen diferencias significativas con P>0.05 entre los promedios observados de los diferentes hatos. Los niveles de infestación promedio en los distintos hatos oscilaron entre 0u y 2lu; donde u= número de huevos y larvas/gramos de heces. De estos 48 promedios (8 hatos por 6 géneros de parásitos), 6 alcanzaron valores >9; 1 hato C1 de la upe Las Brisas con media de 2lu del género Haemonchus, 1 hato e de la upe San Miguel con media de 20.87u del género Strongyloides, 1 hato C2 de la upe San Jorge con media de 17.69u del género Strongyloides, 2 hatos C1 de las upe las Brisas y San Miguel con medias de 10.67u y medias de 10.5u respectivamente, ambos del género Coccidia y 1 hato e2 de la upe San Jorge con media de 9.6lu del género Coccidia. En los promedios restantes se obtuvieron valores <6u perteneciendo a la calificación leve. Posteriormente se realizó un análisis de correlación para determinar el grado de asociación existente entre cada uno de los géneros de parásitos económicamente importantes, encontrándose que el género Coccidia presentó el mayor grado de correlación con respecto a los demás géneros encontrados, se encontró que las correlaciones >= 0.25 corresponden a los géneros Coccidia, Moniezia, Neoascaris, Strongyloides y Haemonchus con r de 0.25, 0.25, 0.48, 0.30 respectivamente, similar para Haemonchus y Moniezia con r de 0.289.

Evaluación de la mortalidad del síndrome de la muerte súbita en condiciones comerciales de la linea genética arbor acres vs. hubbard

La actividad avícola ha alcanzado un gran desarrollo en estos últimos años, pero como toda actividad está sujeta a riesgos o factores que inciden en su labor, en este caso nos referimos al síndrome de muerte súbita (SMS) que desde su aparición se han realizados diversos estudios para encontrar los factores que influyen en la presentación de este síndrome por tal razón en este estudio se evalúa la incidencia del síndrome de muerte súbita en condiciones comerciales en dos líneas como son: Arbor Acres y Hubbard . Con el objetivo de determinar la línea más susceptible en presentar este síndrome como también evaluar las pérdidas económicas que causo este factor en la granja donde se realizó el experimento, tomando en cuenta los parámetros productivos de ambas líneas para lo cual se utilizaron tres galeras de pollos de engorde con una cantidad de 14406,14406 y14295 respectivamente , evaluando las siguientes galeras galera 3, galera 4, y galera 6 equivalente a la suma de 43107 pollos de un día de nacido de la línea arbor acres la galera 3 y 4 representando el tratamiento 1 y de la línea Hubbard la galera 6 representando el tratamiento 2 ,ambos tratamientos con el mismo manejo implementado en les granjas de la empresa TIPTOP . Planteando como variable las siguientes: mortalidad, peso vivo, consumo, conversión alimenticia y ganancia media diaria. Dichos valores obtenidos fueron analizados a través de un diseño estadístico de bloques completos al azar, en el caso de la variable que resultara con significancia estadística se sometió a una prueba de rangos múltiples tukey para encontrar la mejor línea para la actividad de engorde. En el cual el análisis estadístico arrojó los siguientes resultados: Para la variable la mortalidad no se encontró diferencia significativa (p<0.05) entre ambos tratamientos. Al igual que la variable consumo, conversión alimenticia y ganancia media diaria no mostró diferencia significativa (p<0.05) en tanto la variable peso mostró diferencia significativa (p> 0.05) entre ambas líneas , la que fue sometida a la prueba de rango múltiples Tukey en donde se obtuvo que la línea hubbard está más predispuesta a obtener un peso mayor que la línea Arbor Acres. En cuanto a las pérdidas por síndrome de muerte súbita se refiere que la raza hubbard presentó mayor mortalidad por síndrome por muerte súbita , que la raza Arbor Acres, por lo cual la línea hubbard presenta mayores pérdidas económicas aunque cabe mencionar que la línea hubbard presentó una menor mortalidad en general que la línea Arbor Acres y también esta línea ( hubbard) obtuvo un mayor peso vivo que la línea Arbor Acres , indicando que es más viable económicamente explotar esta línea ya que muestra una diferencia de 3.8% entre ambos tratamiento, teniendo un mayor ingreso , el tratamiento 2 (hubbard ). En grandes cantidades este porcentaje es realmente significativo. Como conclusión del presente estudio podemos afirmar lo siguiente la línea más susceptible en presentar el síndrome por muerte súbita es la línea hubbard aunque estadísticamente no muestre diferencia significativa (p< 0.05). Entre los parámetros productivo (variables) como , mortalidad , consumo, conversión alimenticia y ganancia media diaria no existe diferencia estadística , por lo cual ambas líneas poseen similares índice productivo. , Mientras la variable peso muestra diferencia de donde deducimos que la raza hubbard posee un peso mayor que la línea Arbor Acres. En tanto las pérdidas económicas son mayores en el tratamiento 2 (hubbard) y obteniendo en este un mayor rendimiento en producción y en rentabilidad.