Immunophenotyping contribution for the diagnosis of Myelodysplastic Syndrome

Mestrado em em Bioquímica com especialização em Bioquímica ClínicaA Síndrome Mielodisplásica (SMD) constitui um grupo heterogéneo de neoplasias mielóides caraterizados por alterações nos processos de diferenciação e maturação celular. O contributo da citometria de fluxo no diagnóstico da patologia ainda não está bem estabelecido, contudo, desde 2014 o International Leukemia Net European Working Group recomenda a integração da técnica para o diagnóstico destas entidades. Quer a morfologia quer a imunofenotipagem tentam encontrar rasgos displásicos nas linhas maioritárias da medula óssea (MO). Neste contexto, o objetivo deste estudo incidiu na análise das linhas minoritárias da MO como a linha a basófilo, eosinófilo, célula dendrítica plasmacitóide e mastócito, de forma a verificar se estas são fenotipicamente diferentes na hematopoiese normal ou displásica. As amostras analisadas (n=84) encontravam-se distribuídas, de acordo com a classificação da Organização Mundial de Saúde, em: 8 CRDU, 45 CRDM, 17 AREB-1 e 14 AREB-2. Baseado no International Prognostic Scoring System, 18 foram classificadas como de Baixo risco, 19 como de risco Intermédio1 e 16 como de risco Intermédio2-Alto. Paralelamente, analisaram-se 14 MO normais/reativas. Nas diferentes linhas hematopoiéticas estudadas, foi calculada a percentagem de células nos diferentes estadios maturativos e a expressão dos recetores: CD10, CD11b, CD13, CD33, CD34, CD35, CD44, CD45, CD64, CD117, CD123, CD133 e HLA-DR. Procedeu-se à análise da expressão dos genes Triptase e Peroxidase do Eosinófilo para amostras de basófilos e eosinófilos, correspondentemente, em 9 amostras diagnosticadas com SMD e em 5 MO normais/reativas. Observou-se um aumento da expressão de CD44 e CD45 transversal às linhas hematopoiéticas estudadas, um aumento de expressão de CD117 e HLA-DR na linha a mastócito, uma diminuição de CD13 na linha a basófilo e de CD35 e CD64 na linha a eosinófilo. Na análise da expressão génica, observou-se um aumento de expressão dos genes Triptase e Peroxidase do Eosinófilo para as populações de basófilos e eosinófilos, respetivamente, nas amostras da SMD. Os resultados obtidos claramente demonstram padrões alterados nas linhas hematopoiéticas estudadas consistentes com os publicados para a linha granulocítica, monocítica e eritroide, reforçando o contributo da citometria de fluxo para o diagnóstico e estratificação prognóstica dos doentes com SMD.The Myelodysplastic Syndrome (MDS) is a heteregeneous group of myeloid neoplasms characterized by changes in the differentiation and maturation of myeloid cells. The contribution of immunophenotyping for the diagnosis of MDS is not well established, however, since 2014 the International Leukemia Net European Working Group recomends the integration of flow cytometry for the diagnosis of these entities. Either morphology or immunophenotyping try to find dysplastic rips in the majority hematopoietic lineages of bone marrow (BM). In this context, the aim of this study was to look into the BM minority lineages from MDS patients, such as basophil, eosinophil, plasmacytoid dendritic cells and mast cell lineage, in order to verify if they are phenotypically different from normal/reactive bone marrow BM. MDS patients were distributed (n=84), according to the World Health Organization classification as: 8 RCUD, 45 RCMD, 17 RAEB-1 and 14 RAEB-2. Based on the International Prognostic Scoring System, 18 cases were Low risk MDS, 19 were intermediate risk-1 and 16 had classified as intermediate risk-2 and high-risk. Additionally, we analysed 14 normal/reactive BM. In the different hematopoietic lineages, the percentage of cells in different maturation stages and expression of several receptors, like CD10, CD11b, CD13, CD33, CD34, CD35, CD44, CD45, CD64, CD117, CD123, CD133 and HLA-DR were studied. We analysed the Tryptase and Eosinophil Peroxidase gene expression to basophils and eosinophils population, respectively, in 9 MDS samples and in 5 normal/rective BM. There were an increased expression in CD44 and CD45 in all hematopoietic lineages under study, an increased expression in CD117 and HLA-DR in mast cel lineage, a decresead expression in CD13 in basophil lineage and in CD35 and CD64 in eosinophil lineage. In gene expression analysis there were, in MDS samples, an increased expression in Tryptase and Eosinophil Peroxidase genes for basophils and eosinophils populations, respectively. Regarding our results, there were immunophenotypic and functional differences in hematopoietic lineages studied consistent with those published to granulocytic, monocytic and erythroid lineages. This enhancing the contribution of flow cytometry for the diagnosis and prognostic stratification of MDS patients. Índic

Cartografia de Sítios da Rede Natura 2000 em Portugal continental

Resumo: No âmbito da Cartografia de Habitats Naturais e Seminaturais e de Flora em 31 Sítios Classificados formou-se uma equipa capaz de responder adequadamente ao caderno de encargos lançado em Concurso Público em julho de 2018. A metodologia definida seguiu diferentes fases de implementação, preparação de campo; trabalho de campo; execução de cartografia e validação da cartografia. Para a cartografia foi efetuada previamente uma exaustiva preparação do trabalho de campo que contemplou o estudo breve da Paisagem do Sítio e a definição de uma amostragem estratificada com base na cartografia do uso do solo, cartografia anterior de habitats e conhecimento de especialista do SIC. Efetuada a preparação do trabalho de campo, procedeu-se ao início da recolha de dados no campo a qual se focou em polígonos de ocorrência provável de habitats e em polígonos com pontos de amostragem (definidos anteriormente na amostragem estratificada), tendo sido preenchidas ficha de amostragem de habitats, com a respetiva recolha de imagens. O trabalho de campo decorreu entre abril de 2019 e novembro de 2020. Posteriormente ao trabalho de campo, a informação obtida foi organizada e tratada com vista à produção de cartografia, a qual foi organizada numa base de dados geográficos (geodatabase) para garantir a qualidade topológica e temática dos resultados. Os diferentes campos da tabela de atributos apresentavam-se com pré-preenchimento de modo a minimizar a produção de erros e a auxiliar o preenchimento dos atributos da cartografia. Apresenta-se uma síntese dos trabalhos realizados e da informação obtida, com especial enfoque nos habitats que desapareceram e nos que foram agora identificados, para alguns dos Sítios. Para habitats previamente selecionados foram avaliados o grau de conservação e as pressões identificadas, bem como efetuada a análise da representatividade de cada um. De modo transversal e ao longo de todo o processo foram executados procedimentos de controle de qualidade

Practical guide for the use of biological therapies in rheumatoid arthritis - Update of December 2011

The authors review the practical aspects of biologi -cal therapy use for rheumatoid arthritis patients, commenting safety issues before and after treatment initiation and the best treatment strategies to optimize efficacy.publishersversionpublishe

Portuguese guide lines for the use of biological agents in rheumatoid arthritis - october 2011 update

The authors present the revised version of the Portuguese Society of Rheumatology (SPR) guidelines for the treatment of Rheumatoid Arthritis (RA) with biological therapies. In these guidelines the criteria for introduction and maintenance of biological agents are discussed as well as the contraindications and procedures in the case of non-responders. Biological treatment (with a tumour necrosis factor antagonist, abatacept or tocilizumab) should be considered in RA patients with a disease activity score 28 (DAS 28) equal to or greater than 3.2 des pite treatment with at least 20mg-weekly-dose of methotrexate (MTX) for at least 3 months or, if such treatment is not possible, after 3 months of other conventional disease modifying drug or combination therapy. A DAS 28 score between 2.6 and 3.2 with a significant functional or radiological deterioration under treatment with conventional regi -mens could also constitute an indication for biological treatment. The treatment goal should be remission or, if that is not achievable, at least a low disease activity, defined by a DAS28 lower than 3.2,without significative functional or radiological worsening. The response criteria, at the end of the first 3 months of treatment, are a decrease of at least 0.6 in the DAS28 score. After 6 months of treatment res ponse criteria is defined as a decrease greater than 1.2 in the DAS28 score. Non-responders, in accordance to the Rheumatologist's clinical opi -nion, should try a switch to another biological agent (tumour necrosis factor antagonist, abatacept, rituxi mab or tocilizumab).publishersversionpublishe

Characterisation of microbial attack on archaeological bone

As part of an EU funded project to investigate the factors influencing bone preservation in the archaeological record, more than 250 bones from 41 archaeological sites in five countries spanning four climatic regions were studied for diagenetic alteration. Sites were selected to cover a range of environmental conditions and archaeological contexts. Microscopic and physical (mercury intrusion porosimetry) analyses of these bones revealed that the majority (68%) had suffered microbial attack. Furthermore, significant differences were found between animal and human bone in both the state of preservation and the type of microbial attack present. These differences in preservation might result from differences in early taphonomy of the bones. © 2003 Elsevier Science Ltd. All rights reserved

Brazilian Flora 2020: Leveraging the power of a collaborative scientific network

International audienceThe shortage of reliable primary taxonomic data limits the description of biological taxa and the understanding of biodiversity patterns and processes, complicating biogeographical, ecological, and evolutionary studies. This deficit creates a significant taxonomic impediment to biodiversity research and conservation planning. The taxonomic impediment and the biodiversity crisis are widely recognized, highlighting the urgent need for reliable taxonomic data. Over the past decade, numerous countries worldwide have devoted considerable effort to Target 1 of the Global Strategy for Plant Conservation (GSPC), which called for the preparation of a working list of all known plant species by 2010 and an online world Flora by 2020. Brazil is a megadiverse country, home to more of the world's known plant species than any other country. Despite that, Flora Brasiliensis, concluded in 1906, was the last comprehensive treatment of the Brazilian flora. The lack of accurate estimates of the number of species of algae, fungi, and plants occurring in Brazil contributes to the prevailing taxonomic impediment and delays progress towards the GSPC targets. Over the past 12 years, a legion of taxonomists motivated to meet Target 1 of the GSPC, worked together to gather and integrate knowledge on the algal, plant, and fungal diversity of Brazil. Overall, a team of about 980 taxonomists joined efforts in a highly collaborative project that used cybertaxonomy to prepare an updated Flora of Brazil, showing the power of scientific collaboration to reach ambitious goals. This paper presents an overview of the Brazilian Flora 2020 and provides taxonomic and spatial updates on the algae, fungi, and plants found in one of the world's most biodiverse countries. We further identify collection gaps and summarize future goals that extend beyond 2020. Our results show that Brazil is home to 46,975 native species of algae, fungi, and plants, of which 19,669 are endemic to the country. The data compiled to date suggests that the Atlantic Rainforest might be the most diverse Brazilian domain for all plant groups except gymnosperms, which are most diverse in the Amazon. However, scientific knowledge of Brazilian diversity is still unequally distributed, with the Atlantic Rainforest and the Cerrado being the most intensively sampled and studied biomes in the country. In times of “scientific reductionism”, with botanical and mycological sciences suffering pervasive depreciation in recent decades, the first online Flora of Brazil 2020 significantly enhanced the quality and quantity of taxonomic data available for algae, fungi, and plants from Brazil. This project also made all the information freely available online, providing a firm foundation for future research and for the management, conservation, and sustainable use of the Brazilian funga and flora

Growing knowledge: an overview of Seed Plant diversity in Brazil

Abstract An updated inventory of Brazilian seed plants is presented and offers important insights into the country's biodiversity. This work started in 2010, with the publication of the Plants and Fungi Catalogue, and has been updated since by more than 430 specialists working online. Brazil is home to 32,086 native Angiosperms and 23 native Gymnosperms, showing an increase of 3% in its species richness in relation to 2010. The Amazon Rainforest is the richest Brazilian biome for Gymnosperms, while the Atlantic Rainforest is the richest one for Angiosperms. There was a considerable increment in the number of species and endemism rates for biomes, except for the Amazon that showed a decrease of 2.5% of recorded endemics. However, well over half of Brazillian seed plant species (57.4%) is endemic to this territory. The proportion of life-forms varies among different biomes: trees are more expressive in the Amazon and Atlantic Rainforest biomes while herbs predominate in the Pampa, and lianas are more expressive in the Amazon, Atlantic Rainforest, and Pantanal. This compilation serves not only to quantify Brazilian biodiversity, but also to highlight areas where there information is lacking and to provide a framework for the challenge faced in conserving Brazil's unique and diverse flora