Identification of a Rare 3 bp BRAF Gene Deletion in a Thyroid Nodule by Mutant Enrichment with 3'-Modified Oligonucleotides Polymerase Chain Reaction

Papillary thyroid carcinoma (PTC) is the most common malignant thyroid tumor, and 36-69% of PTC cases are caused by mutations in the BRAF gene. The substitution of a valine for a glutamic acid (V600E) comprises up to 95-100% of BRAF mutations; therefore, most diagnostic methods, including allele-specific PCR and real-time PCR, are designed to detect this mutation. Nevertheless, other mutations can also comprise the genetic background of PTC. Recently, a novel and sensitive technique called mutant enrichment with 3'-modified oligonucleotides (MEMO) PCR has been introduced. When we applied allelespecific PCR and MEMO-PCR for the detection of the BRAF V600E mutation, we found an unusual 3' bp deletion mutation (c.1799_1801delTGA) only when using MEMO-PCR. This deletion results in the introduction of a glutamic acid into the B-Raf activation segment (p.V600_K601delinsE), leading to an elevated basal kinase activity of BRAF. This is the first report of a rare 3 bp BRAF deletion in a PTC patient that could not be detected by allele-specific PCR

Doenças crónicas respiratórias

Dissertação de Mestrado em Design com a especialização em Design de Produto apresentada na Faculdade de Arquitetura da Universidade de Lisboa para obtenção do grau de Mestre.O projeto aborda a problemática das doenças crónicas respiratórias, mais especificamente doenças como a asma, alergias e doenças que afetam o foro nasal. Pretendeu-se encontrar uma solução que ajude os pacientes a controlar e a melhorar os sintomas da doença e as condições externas que influenciam a mesma. De uma forma autónoma, os pacientes podem assim aumentar a sua qualidade de vida ao lidarem melhor com a sua doença e ao disponibilizarem informações concretas ao seu médico especialista. Para isso são utilizadas as tecnologias emergentes de robótica e inteligência artificial como ferramenta fundamental para encontrar e desenvolver, em parceria com o design de produto, soluções precisas e personalizadas para uma doença mutável e permanente. Recorrendo à revisão da literatura e à consulta de especialistas, aprofundou-se o conhecimento destas patologias, compreendendo o que são e quais as soluções existentes para estes problemas, bem como a pertinência da atuação do design de produto dentro desta área; averiguou-se, junto de pacientes, quias são as condições, necessidades e dificuldades reais que enfrentam; perceber a evolução no setor da saúde e de que forma se vai transformar com a introdução das novas potencialidades tecnológicas como forma de compreender a doença, o paciente, o mercado e a quais as expectativas do público para este setor. As metodologias de investigação utilizadas incluem revisão literária e recolha de estatísticas, análise de equipamentos similares e de tecnologias aplicáveis nesta especialidade, e inquéritos aos pacientes com estas doenças que irão culminar numa formulação de personas. São também utilizados recursos para a definição de problema, nomeadamente, storyworld, mind map, proposition, stakeholders, swot, storyboard, visão de proposta de valor, business model canvas e blueprint de serviços, assim como compreender o problema e o público, identificar parceiros, recursos disponíveis, áreas de atuação, e definir soluções para gerar novas ideias. Para o desenvolvimento do conceito foram elaborados esquemas, desenhos, e simulações 3D que culminaram num protótipo. Com este projeto desenvolveu-se um equipamento capaz de melhorar a qualidade de vida e o dia-a-dia destes pacientes. Com esse objetivo foram elaboradas soluções de dispositivos inteligentes capazes de compreender a doença e a sua envolvente, melhorando a eficácia do tratamento através de um conjunto de atuações em diversas frentes, elevando o tratamento para lá da solução farmacológica convencional.ABSTRACT: The project addresses the issue of chronic respiratory diseases, such as asthma, allergies and nasal forum diseases. The intention was to find a solution that not only helps patients to control and improve symptoms, but also to improve external conditions that influence it, helping patients to develop and maintain, independently, a better quality of life, improving their relationship with the disease and the relationship with the specialist. For that purpose, the emerging technologies of robotics and artificial intelligence are used as a fundamental tool to find and develop precise and personalized solutions. It is therefore important to deepen the knowledge of these pathologies, understanding what they are and what are the existing solutions, as well as understand the potential of product design within this area. Find out, with patients, what are the real conditions, needs and difficulties they face. Understand the evolution that is taking place in the health sector and how it will evolve with the introduction of new technological potentials, as well as understanding the market and what the public's expectations for this sector are. The research methodologies used include literature review and collection of statistics, survey of similar equipment and technologies applicable to the area and surveys with patients of these diseases that gave rise to the formulation of personas. Problem definition, storyworld, mind map, proposition, stakeholders, swot, storyboard, value proposition vision, business model canvas and service blueprint are also used as a way to understand the problem and the target, identify partners, viable tools, possible areas of action, define solutions and generate ideas. For the development of the concept, schemes, drawings and 3D simulations are elaborated, culminating in a physical model. In this project is was developed an equipment capable of improving the quality and the daily life of these patients. With this objective in mind, solutions are developed for intelligent devices capable of understanding the disease and its surroundings, improving the effectiveness of the treatment through a set of actions on several fronts, elevating the treatment beyond the pharmacological solution.N/

Prevalência de Hipotensão Ortostática na consulta de Cardiologia do Centro Hospitalar Cova da Beira

Introdução: A hipotensão ortostática (OH) - definida como a redução da pressão arterial sistólica> 20 mmHg ou da pressão arterial diastólica> 10 mmHg, após passagem de decúbito dorsal (DD) para posição supina (PS), no período de três minutos. Recomenda-se a medição dos valores de pressão arterial (PA), ao primeiro e terceiros minutos, após adoção da PS. A HO é frequentemente subdiagnosticada, com efeito deletério na morbilidade cardiovascular e mortalidade independente para todas as causas, sendo um marcador de mau prognóstico em pacientes de meia-idade e em idosos. Objetivo: Estudar a prevalência de HO, numa amostra de doentes inscritos na Consulta Externa de Cardiologia do Centro Hospitalar Cova da Beira. Material e Métodos: A recolha de dados realizou-se entre Agosto de 2016 e Março de 2017, através de entrevista presencial, medições dos valores de pressão arterial (PA) e frequência cardíaca (FC), na Consulta Externa de Cardiologia do Centro Hospitalar Cova da Beira, e análise dos processos clínicos. Estipulou-se como critério de inclusão, todos os pacientes com idade> a 55 anos e que informadamente assentiram inclusão no estudo. Os critérios de exclusão incluem pacientes com patologia psiquiátrica que interfira com o processo de decisão livre e informada, pacientes com mobilidade reduzida e pacientes com idade < 55 anos. Resultados: Numa amostra de 87 doentes, com idade média de 72,2±11,8 anos, 59.77% (n=52) dos indivíduos são do sexo masculino (M). A prevalência de HO, na amostra total, para a PA sistólica 26% e de 5% no caso da PA diastólica. Em DD, a PA sistólica média é de de 143,3±21,7 mmHg, a PA diastólica média de 79,1± 10,2 e a FC média de 70,1±13,9 bpm. Ao 3º minuto, a PA sistólica média é de 124,3±37,6 mmHg, a PA diastólica média de 84,3±12,9 mmHg e a FC média de 76,4±18,6 bpm. Da comparação entre os valores (teste t), resulta uma significância de p<0,001 para as variáveis PA sistólica, PA diastólica e FC. Existe correlação entre a PA sistólica DD/PS (R2=0,23); PA diastólica DD/PS (R2=0,39) e entre as respetivas FC (R2=0,67). Conclusão: A HO é frequente, na amostra total, no entanto nos grupos etários superiores, a PA sistólica e a PA diastólica são significativamente mais baixas na PS. Admite-se que a FC sobe compensatoriamente do DD para a PS, sendo este mecanismo consistente na amostra em estudo.Introduction: Orthostatic hypotension – defined as a decrease in systolic blood pressure >20 mmHg or in diastolic blood pressure >10 mmHg, within three minutes from decubitus dorsal (DD) to supine position (SP). The blood pressure (BP) measurement is recommended at the minute 1 and 3 after positioning to SP. OH is often misdiagnosed, with deleterious effect on cardiovascular morbidity and independent mortality for all causes, as a bad prognosis mark on middle-age and elder patients. Purpose: To study the OH prevalence on a sample of patients enrolled in outpatient cardiology service of Centro Hospitalar Cova da Beira. Material and Methods: Data were collected between August 2016 and March 2017 through personal interview, blood pressure (BP) and heart rate (HR) measurements at outpatient cardiology service at Centro Hospitalar Cova da Beira and clinical processes assessment. The inclusion criterion was established as all patients > 55 years old who had given informed consent to take part in the study. The exclusion criteria include patients with psychiatric pathology that may compromise the informed and free consent process, patients with reduced mobility and patients < 55 years old. Results: On a sample with 87 patients, with mean age 72.2±11.8 years, 59.77% (n=52) of the subjects are male (M). The prevalence of OH, on total sample, for systolic BP is 26% and 5% for diastolic BP. On DD, mean systolic BP is 143.3±21.7 mmHg, mean diastolic BP 79.1± 10.2 and mean HR 70.1±13.9 bpm. On minute 3, mean systolic BP is 124.3±37.6.7 mmHg, mean diastolic BP 84.3± 12.9 and mean HR 76.4±18.6 bpm. From values comparison (t test) results a p<0.001 significance for systolic BP, diastolic BP and HR variables. There is a correlation between systolic BP DD/PS (r2=0.239; diastolic BP DD/PS (R2=0.39) and associated HR (R2=0.67). Conclusion: OH is frequent on total sample; however, on older age groups, systolic BP and diastolic BP are significantly lower on PS. It is considered that HR increases accordingly from DD to PS, and this mechanism is consistent on the study sample

A transposição da narrativa da performance ao audiovisual

Mestrado em Comunicação MultimédiaO presente trabalho propõe-se fazer um estudo comparativo de duas obras: À Espera de Godot, de Samuel Beckett, e A Espera, de Helena Botto. A segunda consiste numa performance adaptada da primeira. Ambas se inserem numa tipologia de teatro designado Teatro do Absurdo e como tal, torna-se necessário fazer uma contextualização e análise do mesmo. Sendo a performance de Helena Botto uma adaptação da referida obra de Beckett, este trabalho contempla uma secção dedicada ao estudo deste fenómeno, destacando a questão da fidelidade à obra adaptada. Numa segunda fase, este trabalho procura estudar a transposição da performance de Helena Botto para filme, bem como, a possibilidade de um público específico fazer a visualização do mesmo de forma interactiva, mais concretamente segundo uma ordem cronológica diferente da original. ABSTRACT: The following work proposes a comparative study of two plays: Waiting for Godot, by Samuel Beckett, and A Espera, by Helena Botto. The latter consists of a performance adapted from the former play. Both are encompassed by the theatrical concept of The Theatre of the Absurd and, as such, the need arises to contextualize and analyze this concept. Due to the fact that the work of Helena Botto is an adaptation of Beckett’s, this study includes a section dedicated to the understanding of this phenomenon, questioning whether or not the adapted work is faithful to its original rendition. Following this first phase, this work aims to study the transposition of Helena Botto’s performance into film, as well as the possibility of an interactive visualization by a given audience, using a different chronological timeline than the original work

Relatório de estágio II de Ana Margarida Bento Trovisco

Para a conclusão do Mestrado Integrado em Ciências Farmacêuticas fez-se cumprir o estágio curricular com a duração de cinco meses. Este relatório reflete, de forma sucinta, o conhecimento e aprendizagem adquiridos tanto a nível de gestão - numa componente mais organizacional, seja de aconselhamento ao doente, dispensa de medicamentos e cuidados de saúde prestados. O contacto com o utente, foi sem dúvida, o maior desafio desta componente prática. O respetivo relatório encontra-se dividido em duas partes, sendo que na primeira parte é descrito, de forma sucinta, o funcionamento da Farmácia Ferreira da Silva, todo o back office e ato de dispensação. Numa segunda parte são descritas atividades por mim realizadas ao longo do estágio, entre elas a implementação de Kaizen Lean – melhoria contínua e a elaboração de uma ação de formação sobre Psoríase aliada à promoção da marca Iralfaris- ISDIN

BRAFV600E Mutation and p27kip1 Expression in Papillary Carcinomas of the Thyroid ≤1 cm and Their Paired Lymph Node Metastases

Abstract BACKGROUND. BRAFV600E mutation and p27kip1 expression have been introduced as novel indicators that may predict prognosis in different tumors, as well as in papillary thyroid carcinomas. METHODS. Tissue samples from 214 consecutive patients who underwent total or near-total thyroidectomy with histological diagnosis of papillary thyroid carcinoma (PTC) ≤1 cm were analyzed for BRAFV600E mutation by a real-time, allele-specific amplification and for p27kip1 expression by immunohistochemistry. RESULTS. The BRAFV600E mutation was detected in 88 of the tumors examined, with significant differences between groups with and without lymph node (LN) metastases; the mean age of patients with BRAF V600E mutation was significantly higher than that of patients without mutations. A significant association was found between low p27Kip1 protein expression and multifocality, bilaterality, and extrathyroidal extension, in addition to LN metastasis. In 42 cases with LN metastases, 23 harbored the BRAFV600E mutation in the metastatic tumor and presented a wider diameter of the largest metastatic area, a higher number of involved LNs, and a higher percentage of metastatic lesions with extracapsular extension of LN (ECE-LN). A significantly lower mean value of p27Kip1 was observed in LNs harboring the BRAFV600E mutation and in ECE-LN; an inverse correlation was found between p27Kip1 and the number of metastatic LNs, as well as the diameter of the largest metastatic area in LN. CONCLUSIONS. The authors' data suggested that BRAFV600E mutation and p27Kip1 down-regulation in cancer cells of PTC <1 cm may be factors that facilitate tumor-cell growth and progression once these are seeded in the LNs. © 2007 American Cancer Society

FlyBase 2.0: the next generation.

FlyBase (flybase.org) is a knowledge base that supports the community of researchers that use the fruit fly, Drosophila melanogaster, as a model organism. The FlyBase team curates and organizes a diverse array of genetic, molecular, genomic, and developmental information about Drosophila. At the beginning of 2018, 'FlyBase 2.0' was released with a significantly improved user interface and new tools. Among these important changes are a new organization of search results into interactive lists or tables (hitlists), enhanced reference lists, and new protein domain graphics. An important new data class called 'experimental tools' consolidates information on useful fly strains and other resources related to a specific gene, which significantly enhances the ability of the Drosophila researcher to design and carry out experiments. With the release of FlyBase 2.0, there has also been a restructuring of backend architecture and a continued development of application programming interfaces (APIs) for programmatic access to FlyBase data. In this review, we describe these major new features and functionalities of the FlyBase 2.0 site and how they support the use of Drosophila as a model organism for biological discovery and translational research

Mutation analysis of B-RAF gene in human gliomas

The RAS/RAF/MEK/ERK kinase pathway is pivotal in the transduction of mitogenic stimuli from activated growth factor receptors, which regulates cell proliferation, survival, and differentiation. Up-regulation of this pathway due to RAS mutations is found in approximately 30% of human tumors. Recently, activating mutations of B-RAF were identified in a large proportion of human cancers. Gliomas are the most frequent primary central nervous system tumors and the molecular mechanisms that underlie the development and progression of these tumors are far from being completely understood. The purpose of this study was to clarify the incidence of B-RAF mutations and their possible relation with tumor progression in a series of 82 human gliomas, including 49 astrocytic and 33 oligodendroglial tumors. The analysis of B-RAF hotspot regions, exons 11 and 15, showed presence of B-RAF mutations in only 2 out of 34 (6%) glioblastomas, and absence in the remaining histological types. Both mutations were located in the hotspot residue 600 (V600E) at exon 15, which leads to constitutive B-RAF kinase activity. These data suggest that activating mutations of B-RAF are not a frequent event in gliomas; nevertheless, when present they are associated with highgrade malignant lesions