Genetic susceptibility to myeloproliferative neoplasms and therapeutic efficacy

RESUMO: As neoplasias mieloproliferativas são classicamente divididas em neoplasias mieloproliferativas BCR-ABL1 (cromossoma Philadelphia) positivas, como é o caso da leucemia mieloide crónica, e BCR-ABL1 negativas, incluindo a policitémia vera, a trombocitémia essencial e a mielofibrose primária. Estas entidades nosológicas têm origem na transformação maligna das stem-cells hematopoiéticas, levando a amplificação anormal e à proliferação das células das linhagens mielóides. A diversidade fenotípica das neoplasias mieloproliferativas Philadelphia negativas resulta da combinação de mutações somáticas já identificadas e caracterizadas, variabilidade genética hereditária, regulação pós-genética e condicionantes individuais. De acordo com a literatura, vários polimorfismos genéticos foram identificados, podendo influenciar os mecanismos de apoptose e de capacidade de reparação do DNA, comprometendo a transcrição genética e/ou a atividade celular, conferindo predisposição genética para a doença, incluindo o caso das neoplasias mieloproliferativas, e condicionando a resposta terapêutica, a evolução clínica e o prognóstico. O presente trabalho teve como objetivo caraterizar a população portuguesa de acordo com o tipo de neoplasia mieloproliferativa Philadelphia negativa, a sua prevalência e a presença da mutação JAK2, dando especial ênfase a alguns doentes em particular; investigar o papel dos polimorfismos nos genes envolvidos nas vias de apoptose e de reparação por excisão de bases na susceptibilidade para estas doenças e, juntamente com outros fatores de risco, na sobrevivência, evolução clínica e prognóstico. Foram realizados estudos caso-controlo numa amostragem de 133 doentes Portugueses caucasianos e 281 indivíduos saudáveis correspondentes e foram selecionados nove e oito polimorfismos de genes envolvidos nas vias de apoptose e de reparação por excisão de bases, respectivamente, que foram genotipados utilizando a técnica de Polymerase Chain Reaction em tempo real. A análise estatística foi realizada com recurso ao software SPSS versão 22.0. Os resultados deste estudo revelaram uma distribuição dos doentes por patologia de 60,2% com trombocitémia essencial, 29,3% com policitémia vera e 10,5% com mielofibrose primária, um discreto predomínio nas mulheres e 75,0% dos doentes positivos para a presença da mutação JAK2 V617F, evidenciando uma incidência mais elevada na trombocitémia essencial e mais reduzida na policitémia vera, relativamente ao esperado. Entre os doentes, alguns casos em particular foram destacados e apresentados nesta tese: o caso de um doente com policitémia vera JAK2 exão 12 positivo, apresentando uma nova mutação - c.1605G> T (p.Met535Ile) - associada a outra mutação já anteriormente descrita, evidenciando um fenótipo clínico atípico; dois doentes com trombocitémia essencial, com suspeita da coexistência rara da mutação JAK2 V617F e da translocação BCR-ABL1; e um doente com leucemia mieloide crónica,apresentando uma nova mutação no domínio kinase BCR-ABL1 - c.839T> G (p.Val280Gly) - que poderá estar associada a resistência ao imatinib. Quanto aos polimorfismos investigados, os nossos resultados revelaram potenciais associações entre alguns dos polimorfismos e a susceptibilidade individual para estas doenças, sugerindo o potencial envolvimento de polimorfismos do gene CASP9 (Phe136Phe) e dos genes XRCC1_399 (Gln399Arg) e MUTYH (Gln335His), após estratificação por patologia para os doentes com trombocitémia essencial e quando se tratam de mulheres. Nos doentes JAK2 positivos verificou-se uma associação com os polimorfismos do gene CASP9 (Phe136Phe) e do gene XRCC1_399 (Gln399Arg). Nos genes de reparação por excisão de bases, a combinação de alelos também demonstrou associação com doença para um haplogrupo específico. Em relação à evolução clínica dos doentes sob tratamento, na maioria dos casos com hidroxiureia, de acordo com nossos resultados: 17 doentes apresentaram progressão para mielofibrose secundária/leucemia mieloide aguda, influenciando a sobrevivência e estando associada à exposição a agentes citoredutores. Nestes doentes, houve também evidência global de associação com polimorfismos do gene CASP8 (3'UTR) e do gene XRCC1_194 (Arg194Trp), e após estratificação para trombocitémia essencial com polimorfismos do gene CASP9 (Arg173His) e dos genes APEX1 (Asp148Glu) e XRCC1_194 (Arg194Trp); 11 doentes desenvolveram uma neoplasia não mieloide primária de novo, evidenciando uma associação global com polimorfismos do gene CASP8 (Asp270His) e do gene XRCC1_399 (Gln399Arg); 22 doentes apresentaram eventos trombóticos e uma associação global com polimorfismos do gene XRCC1_399 (Gln399Arg). Os resultados apresentados na presente dissertação ajudaram a caracterizar uma população de doentes com neoplasias mieloproliferativas Philadelphia negativas, podendo refletir a realidade portuguesa, e revelaram o papel potencial de determinados polimorfismos e outros fatores na susceptibilidade para a doença e na evolução clínica dos doentes sob terapêutica, com impacto prognóstico. Embora sejam necessários estudos mais alargados para confirmar estes resultados, estes novos dados podem contribuir para um melhor conhecimento da fisiopatologia e, no futuro, para uma escolha mais racional e eficiente das estratégias terapêuticas a adotar nestas patologias.ABSTRACT: Myeloproliferative neoplasms are classically divided into BCR-ABL1 (Philadelphia chromosome) positive chronic myeloid leukemia and BCR-ABL1 negative myeloproliferative neoplasms, including polycythemia vera, essential thrombocythemia and primary myelofibrosis. These disorders have origin from the malignant transformation of a hematopoietic stem-cell, leading to abnormal amplification and proliferation of myeloid lineages. The phenotypic diversity of Philadelphia chromosome negative myeloproliferative neoplasms results from the combination of somatic mutations already identified and characterized, inherited genetic variability, postgenetic regulation, and individual conditioners. According to the literature, several genetic polymorphisms have been identified, which may influence the apoptosis mechanisms and the DNA repair capacity, compromising genetic transcription and/or cell function, conferring genetic predisposition to disease, including myeloproliferative disorders, and conditioning the therapeutic response, the clinical outcome and prognosis. The present work had the purpose of characterize the Portuguese population according to the type of Philadelphia chromosome negative myeloproliferative neoplasm, its prevalence and the presence of JAK2 mutation, with special emphasis on some particular patients; investigate the role of polymorphisms in genes involved in apoptosis and base excision repair pathways in these disorders susceptibility and, along with other risk factors, in survival, clinical outcome and prognosis. Case-control studies were carried out in a Caucasian Portuguese sample of 133 patients and 281 matched control subjects, nine and eight polymorphisms of genes involved in apoptosis and base excision repair pathways, respectively, were selected and genotyped using real time polymerase chain reaction technique and statistical analysis was performed with SPSS version 22.0. The results from this study revealed a pathology distribution of 60.2% of patients with essential thrombocythemia, 29.3% with polycythemia vera and 10.5% with primary myelofibrosis, with a discrete predominance in females, and a total of 75.0% of patients positive for the presence of the JAK2 V617F mutation, with an increased incidence in essential thrombocythemia and a decreased incidence in polycythemia vera patients than expected. Among the patients, some particular cases were highlighted and presented in this thesis, namely the case of a polycythemia vera patient JAK2 exon 12 positive, presenting a novel mutation – c.1605G>T (p.Met535Ile) - associated with another mutation previously described, evidencing an atypical clinical phenotype; two patients with essential thrombocythemia, with suspicion of the rare coexistence of JAK2 V617F mutation and BCR-ABL1 translocation; and a patient with chronic myeloid leukemia, presenting a new BCR-ABL1 mutation kinase domain mutation – c.839T>G (p.Val280Gly) – which might be associated with resistance to imatinib. Concerning the polymorphisms investigated, our results revealed potential associations between some polymorphisms and individual susceptibility to these disorders, suggesting the potential involvement of CASP9 (Phe136Phe) gene and XRCC1_399 (Gln399Arg) and MUTYH (Gln335His) genes polymorphisms after stratification by pathology diagnosis for essential thrombocythemia patients and when they are women. When patients are JAK2 positive an association with CASP9 (Phe136Phe) gene and XRCC1_399 (Gln399Arg) gene polymorphisms was found. In base excision repair genes, combination of alleles also demonstrated an association with the disease for one specific haplogroup. Regarding the clinical outcome in patients under treatment, the majority of them with hydroxyurea, according to our results: 17 patients showed progression to secondary myelofibrosis/acute myeloid leukemia, influencing survival and being associated with exposure to cytoreductive agents. In these patients, there was also global evidence of association with CASP8 (3’UTR) gene and XRCC1_194 (Arg194Trp) gene polymorphisms, and after stratification for essential thrombocythemia with CASP9 (Arg173His) gene, and APEX1 (Asp148Glu) and XRCC1_194 (Arg194Trp) genes polymorphisms; 11 patients developed a new primary nonmyeloid neoplasm, evidencing a global association with CASP8 (Asp270His) gene and XRCC1_399 (Gln399Arg) gene polymorphisms; 22 patients presented thrombotic events and a global association with XRCC1_399 (Gln399Arg) gene polymorphisms. The results shown in the present dissertation helped to characterize a population of Philadelphia negative myeloproliferative neoplasms patients that could reflect Portuguese reality, and have revealed the potential role of polymorphisms and other factors in disease susceptibility and clinical outcome of patients under treatment, with prognostic impact. Although larger studies are required to confirm these results, these new data may contribute to a best knowledge of the pathophysiology and, in the future, to a more rational and efficient choice of therapeutic strategies to be adopted in these disorders

Factor VIII deficiency inhibition or abnormal contact

Poster presented at the International Society on Thrombosis and Haemostasis 2015 Congress, 20-25 June 2015, Toronto

Rivaroxabano e actividade anti-Factor X activado

Poster apresentado no 5º Simpósio de Tromboembolismo: Progressos na Profilaxia e Terapêutica do Tromboembolismo, Janeiro 2015, Lisboa

Chlorophyll-a and the Supply Side Ecology: Lessons from the Rocky Shores

The aims of this study were to summarize and describe the influences of phytoplankton on the larval cycle of rocky shore invertebrates, and to assess the relationship between fluctuations in chlorophyll-a concentration and the rates of larval processes. We carried out a mini review of the published data regarding the theme of the chapter, in which we described the ecological trends for the most common taxa and key species at small and larger spatiotemporal scales. The following topics were addressed: (i) the influence of phytoplankton on larval development, rhythms of larval release, larval quality, larval transport, settlement, and recruitment; (ii) the relationships between variations in chlorophyll-a concentration and the rates of larval processes; (iii) climate change on phytoplankton larva dynamics. The information presented here highlights the role of phytoplankton on rocky shore communities, as well as the importance of chlorophyll-a as a tool for modeling and forecasting the supply side ecology in rocky shore communities

The active mathematics project at ISCAP

This paper summarizes a project that is contributing to a change in the way of teaching and learning Mathematics. Mathematics is a subject of the Accounting and Administration course. In this subject we teach: Functions and Algebra. The aim is that the students understand the basic concepts and is able to apply them in other issues, when possible, establishing a bridge between the issues that they have studied and their application in Accounting. As from this year, the Accounting course falls under in Bologna Process. The teacher and the student roles have changed. The time for theoretical and practical classes has been reduced, so it was necessary to modify the way of teaching and learning. In the theoretical classes we use systems of multimedia projection to present the concepts, and in the practical classes we solve exercises. To supplement our theoretical and practical classes we have developed an active mathematics project called MatActiva based on the Moodle platform offered by PAOL - Projecto de Apoio Online (Online Support Project). In the last versions of Moodle, it is possible use the TeX language to create math questions. Using this tool we created a set of interactive materials. With the creation of this new project we wanted to take advantage already obtained results with the previous experiences, giving to the students opportunities to complement their study in Mathematics. One of the great objectives is to motivate students, encourage them to overcome theirs difficulties through an auto-study, giving them more confidence and the opportunity to seeing others perspectives of the mathematics subjects. In the MatActiva project the students have a big collection of information about the way of the subject works, which includes the objectives, the program, recommended bibliography, evaluation method and summaries. It works as material support for the practical and theoretical classes, the slides of the theoretical classes are available, the sheets with exercises for the students to do in the classroom and complementary exercises, as well as the exams of previous years. Students can also do diagnostic tests and evaluation tests online. Our approach is a reflexive one, based on the professional experience of the teachers that explore and incorporate new tools of Moodle with their students and coordinate the project MatActiva

Teaching and learning mathematics using moodle

This paper summarizes a project that is contributing to a change in the way of teaching and learning Mathematics. Mathematics is a subject of the Accounting and Administration course. In this subject we teach: Functions and Algebra. The aim is that the student understand the basic concepts and is able to apply them in other issues, when possible, establishing a bridge between the issues that they have studied and their application in Accounting. As from this year, the Accounting course falls under in Bologna Process. The teacher and the student roles have changed. The time for theoretical and practical classes has been reduced, so it was necessary to modify the way of teaching and learning. In the theoretical classes we use systems of multimedia projection to present the concepts, and in the practical classes we solve exercises. We also use the Excel and the mathematical open source software wxMaxima. To supplement our theoretical and practical classes we have developed a project called MatActiva based on the Moodle platform offered by PAOL - Projecto de Apoio Online (Online Support Project). With the creation of this new project we wanted to take advantage already obtained results with the previous experiences, giving to the students opportunities to complement their study in Mathematics. One of the great objectives is to motivate students, encourage them to overcome theirs difficulties through an auto-study giving them more confidence. In the MatActiva project the students have a big collection of information about the way of the subject works, which includes the objectives, the program, recommended bibliography, evaluation method and summaries. It works as material support for the practical and theoretical classes, the slides of the theoretical classes are available, the sheets with exercises for the students to do in the classroom and complementary exercises, as well as the exams of previous years. Students can also do diagnostic tests and evaluation tests online. Our approach is a reflexive one, based on the professional experience of the teachers that explore and incorporate new tools of Moodle with their students and coordinate the project MatActiva

Avaliação da aplicabilidade do instrumento de classificação do paciente pediátrico na perspectiva do domínio família

Trata-se de um estudo piloto descritivo e exploratório, que teve como objetivo geral analisar a aplicabilidade do Instrumento de Classificação do Paciente Pediátrico versão Beta (ICPP – versão Beta) de (DINI; GUIRARDELLO, 2014) em relação aos cuidados de enfermagem, com foco no domínio família. Este estudo foi realizado em duas unidades de internação pediátrica do Hospital de Clínicas de Porto Alegre. Os participantes foram 40 pacientes com idade entre 1 mês e 13 anos. A coleta dos dados deu-se por meio da aplicação do ICPP-versão Beta e a análise dos dados provenientes desta aplicação foi realizada com auxílio dos programas Microsoft Excel® e SPSS 20.0® for Windows. O projeto foi aprovado pela Comissão de Pesquisa da Escola de Enfermagem da Universidade Federal do Rio Grande do Sul (COMPESQ) e pelo Comitê de Ética do Hospital de Clínicas de Porto Alegre. Os resultados do estudo apontam que o instrumento de classificação do paciente pediátrico se aplica nas unidades em que foi realizado o estudo, no que tange o domínio família, sendo viável como medida de acompanhamento do grau de dependência dos pacientes, demonstrando alta confiabilidade e consistência interna das respostas obtidas nas unidades pesquisadas. No campo onde foi aplicado mede consistentemente a característica referente ao domínio família, além de demonstrar a importância do envolvimento do acompanhante no cuidado à criança hospitalizada, bem como sua disponibilidade para o aprendizado de habilidades

Sítios de cultivo

The poetic production here contemplated presents the compilation of works based on everyday practices and experimental doing, involving cultivation and care related to gardening. However, it reverberates in enlarged forms of the concept. Within this perspective, I compose gardens in the context of art as a personal, collective, emotional and artistic activity. Seeking the use of different devices - vases, books, videos, continents that enable cultivation practices as well as openness to new senses, that can be recognized when repracticing ordinary actions. To do so, I dwell on theoretical as Martin Heidegger and Alfredo Bosi, approaching the word cultivation, Anne Cauquelin concerning the garden, Michel of Certeau, on the everyday life and Reinaldo Laddaga addressing about the art, and also in the production of artists like Claude Monet, Maria Ivone dos Santos and Hélio Fervenza, Marilá Dardot, Paulo Damé, Fernando Limberger, who led to the reflection of the creation processes, which is based on everyday life activities, and the cultivation of plants as art.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESA produção poética aqui contemplada, apresenta a compilação de trabalhos realizados a partir de práticas cotidianas e do fazer experimental, envolvendo cultivo e cuidado ligados à jardinagem. Entretanto, reverberando em formas mais alargadas do conceito. Dentro desta perspectiva, componho jardins no contexto da arte como atividade pessoal, coletiva, afetiva e artística. Buscando na utilização de diferentes dispositivos - vasos, livros, vídeos, continentes que permitam práticas de cultivo e a abertura para novos sentidos, que podem ser atribuídos ao repraticar ações ordinárias. Para tanto, me debruço sobre teóricos como Martin Heidegger e Alfredo Bosi, versando sobre a palavra cultivo, Anne Cauquelin acerca do jardim, Michel de Certeau, sobre o cotidiano e Reinaldo Laddaga abordando sobre a arte, e também na produção de artistas como Claude Monet, Maria Ivone dos Santos e Hélio Fervenza, Marilá Dardot, Paulo Damé, Fernando Limberger, que conduziram a reflexão dos processos de criação, que tem como fundamento as atividades do cotidiano, e o cultivo das plantas como arte