Genome editing to be applied in functional characterization of genetic variants in familial breast cancer patients

RESUMO: O cancro da mama (CM) familiar representa 15% de todos os casos de CM, estando principalmente relacionado a variantes hereditárias nos genes BRCA. Vários genes ligados ao CM estão sobretudo relacionados com a Recombinação Homóloga (RH), a principal via de reparação de quebras de cadeia-dupla de DNA. A presença de variantes genéticas nestes genes, como o BRCA1 e BRCA2, pode comprometer a RH e consequentemente o desenvolvimento da doença. No cancro da mama familiar, o aconselhamento genético e a triagem através de testes genéticos têm se tornado padrão, aumentando a deteção de variantes de significado desconhecido (VUS). O efeito biológico atribuído às VUS é pouco claro, sendo necessários ensaios funcionais para caracterizar o seu significado clínico. Num estudo anterior, uma VUS foi identificada no gene BRCA1 em pacientes saudáveis, mas com histórico familiar de cancro relevante. Para estudar a sua relevância funcional, implementamos um modelo in vitro com linhas celulares de mama (MCF10-A e MCF-7) para introduzir a VUS de interesse com uma ferramenta de edição genómica, CRISPR-Cas9 e avaliar a resposta celular através de um desafio genotóxico com doxorrubicina (DOX). Introduzimos com sucesso a VUS como clone heterozigoto nas MCF10-A. Várias metodologias foram selecionadas para avaliar e comparar a resposta celular às lesões genéticas: ensaios do Cometa, ɣ-H2AX e Anexina V. Adicionalmente, avaliamos a expressão relativa da proteína através de Western Blot. Os resultados do ensaio do cometa mostram uma diminuição da sensibilidade à DOX no MCF10-A VUS, porém, no ensaio ɣ-H2AX, observamos um maior % de quebras e cadeia-dupla. Na Anexina V, o MCF10- A VUS apresentou menor % de células em necrose. Por último, a expressão da proteína BRCA1 encontra-se diminuída nas MCF10-A VUS. No geral, os resultados mostram uma diminuição da suscetibilidade à DOX para a linha celular com a VUS, sugerindo um efeito benigno. No entanto, são necessários mais ensaios funcionais para entender o seu papel no risco de cancro.ABSTRACT: Familial breast cancer (BC) is responsible for 15% of all BC cases being mainly linked to inherited variants in BRCA genes. Several genes associated with BC are mostly related to Homologous Recombination (HR), the main pathway to repair DNA double-strand breaks. The occurrence of genetic variants in these genes, such as BRCA1 and BRCA2, might compromise the HR, thus the development of disease. In familial breast cancer the genetic counselling and the screening through genetic testing has been widespread increasing detection of variants of unknown significance (VUS). The biologic effect attributed to those VUS is mostly unclear, so functional assays need to be performed to characterize their mutational status. In a previous study a VUS was identified in the BRCA1 gene in healthy patients but with a relevant familial history of cancer. To study its functional relevance, we implemented an in vitro model using breast cell lines (MCF10-A and MCF-7). to introduce the VUS of interest with a genome editing tool, CRISPR-Cas9 and assess the cellular response through a genotoxic challenge with doxorubicin (DOX). We successfully established MCF10-A heterozygous clone for the VUS. Several methodologies were selected to evaluate and compare the cellular response to genetic lesions: Comet, ɣ-H2AX and Annexin V assays. Also, we assessed the protein relative expression using Western Blot. The comet assay results showed a decreased sensitivity to DOX in MCF10-A VUS, yet, in ɣ-H2AX assay, we observed a higher % DSB. In Annexin V, MCF10-A VUS showed lower % of cells in necrosis. Lastly, the expression of BRCA1 protein was decreased in MCF10-A VUS. Overall, the results show a decreased susceptibility to DOX for the VUS cell line, suggesting a benign behaviour. Nonetheless more functional assays need to be performed to understand their role on cancer risk

Looking for the Core of a Knowledge-based Sea Cluster: A Social Network Analysis in a Maritime Region

For more than two decades cluster policies have emerged as a central focus for decision-makers trying to instigate territorial development. The benefits, especially in terms of collective learning, knowledge sharing and other types of agglomeration economies and spill-over effects, are well stressed in the regional science literature. Today the relevance of maritime activities and marine resources to economic development is acknowledged. For several European countries, the Atlantic Ocean is part of their common history, identity and potential for developing advanced economic niches of excellence. There is no surprise that several regions are trying to implement their development strategies based on a broad Sea Cluster notion that encompasses a diversity of economic activities such as fisheries and aquaculture, coastal tourism, marine transports and activities based on marine sciences and maritime technologies. Based on the results of a trans-regional evaluation performed for the Atlantic Area under project KIMERAA, this paper evaluates the consolidation of the Sea Cluster in the Algarve, a Portuguese region internationally known by its coastal tourism. The region has also been experiencing a growing capacity in economic activities linked to marine sciences. This regional cluster did not emerge spontaneously and there are several initiatives to promote it. Interviews to regional actors showed light on two important issues. i) Which organization should be the main mediator to bridge science to market? ii) Who is the actor that is in a better position to assume a pivotal role in the formal consolidation of the cluster? Using social network analysis the main knowledge transfer mediator and the central actors are identified. Their roles and specific policy implications are underlined.

Caracterização de Puccinia hemerocallidis causadora do primeiro surto de ferrugem de lírio-de-um-dia na Europa

Daylily (Hemerocallis spp.) is an ornamental plant widely used in gardens. Daylily rust, caused by the fungus Puccinia hemerocallidis, has disseminated through all continents only in the 21th century, except in Europe, where it has been considered a quarantine disease by the European Plant Protection Organisation. In Portugal, since November 2015, typical rust symptoms were observed in daylily plants in gardens in Lisbon, Alentejo, Algarve and Madeira, attaining high prevalence, incidence and severity. The causal agent was identified as P. hemerocallidis and the Koch’s postulates were fulfilled. Phylogenetic data suggest that this fungus may have been introduced from North America. Using flow cytometry, the genome size of the P. hemerocallidis populations present in Portugal was estimated to be 345 Mbp (0.3533 pg DNA/1C). For such analysis Rhamnus alaternus was validated as a DNA standard, exhibiting a nuclear content of 0.680 pg DNA/2C. The identification of this disease in diverse locations in Portugal represents a threat to European breeding and nursery industries, since there are the appropriate conditions for inoculum maintenance and propagation from Portugal to the rest of Europeinfo:eu-repo/semantics/publishedVersio

Factors associated with motivation to avoid meeting family and friends during the COVID-19 pandemic

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Factors associated with avoidance of emergency department visits in Portugal during the pandemic

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Smart modular dispenser for medication administration

Over the years, average life expectancy tends to increase, which directly translates into an increase of the aging population. Consequences of this process are the decline or loss of cognitive and motor capacities, reflecting directly on the quality of life and the autonomy of the people, compromising the fulfillment of a pharmacological plan. It is known that, inevitably, medicines are increasingly introduced as part of human life in order to improve the quality of life. However, especially in the elderly and people with dementia, the difficulties in strictly complying with their medicine intake are increasingly evident. In order to counteract this situation, this article presents a device that assists them in this challenge, overcoming situations of overdose, drug interactions and even forgetfulness, due to the lack of recall of instructions, doses and schedules. Although there are different types of pills dispensers on the market, the disclosure herein includes a number of novel features. It is a small and portable pill dispenser so that it is easy to carry. It is able to adapt to the pharmacological needs of the individual since it is modular. It presents audible and luminous alerts in order to facilitate their use and interpretation. This device is mostly used for pills of various shapes and sizes, but is also intended to be capable of holding drops or powdered medication.info:eu-repo/semantics/publishedVersio

UPTAKE ICT: A NETWORK OF STAKEHOLDERS AGAINST DIGITAL ILLITERACY

#Uptake_ICT2life-cycle: digital literacy and inclusion to learners with disadvantaged background# is an Erasmus + project that aims at enhancing digital literacy among adults with disadvantaged backgrounds. The partners have produced didactic materials and pedagogical guidelines to meet this aim. Based on these materials, and their didactical exploitation, a network of stakeholders was formed and trained in order to subsequently be able to teach citizens that both have disadvantaged backgrounds and are digitally illiterate or quasi-illiterate. This paper relates how this experience was undertaken in Portugal in what concerns the creation of the Stakeholders' network and presents its results. The reality that has boosted the creation of this project was the shocking situation portrayed in 2013 Eurostat statistics, according to which one in every three Portuguese had never used the Internet. Uptake ICT was then conceived and designed in order to engage synergies to counter this problem, aiming at a variety of focus groups (but paying special attention to learners of various ages that have never used Internet, like adults with disadvantaged backgrounds), in line with the transversal priorities for education, training and youth of the European Commission and seeking to assist in the meeting of the aims of Europe 2020. To add up, it also intended to enhance and to develop ideas that answer to the Societal Challenges’ needs, by sharing and creating scientific, social, technological and policies impact. The main aspects that the project focused on were digital literacy inclusion, re-qualification and employability of disadvantaged citizens, in order to help them to face the present process of civilizational change (social, political, economic, and cultural). The addressed priorities were to contribute towards a reduction of the number of low-skilled adults (re-skilling and up-skilling of adults thanks to lifelong-learning and training), and to strengthen the links between education and employment in the area of ICT | New technologies | digital literacy and digital competences | basic skills. After having identified both the most preeminent needs and ways of integrating ICT in daily life, and a set of good practices already tested in the areas of digital literacy, inclusion and employability, the project team has built a number of educational contents addressing the issues that were considered most relevant in three main levels of knowledge (Basic, Intermediate, and advanced) , and in the four languages of the project (Portuguese, Italian, English and German). The decision to work on the three levels was due to the fact that in the various partner countries there were groups of target audiences that were positioned differently with respect to their level of digital expertise. The teachinglearning materials that were conceived were afterwards reworked in order to fit in a variety of contexts and formats (e-modules, ebook, MOOC). This option for multi-format was taken having in mind different learning profiles, and the need to provide flexible and attractive materials in order to avoid any kind of rejection. Finally, a number of didactical guidelines were produced in order to provide an interface of suggestions to the stakeholders that would use these materials in their classes or workshop sessions.info:eu-repo/semantics/publishedVersio

systematic review

Publisher Copyright: ©Ana Soraia Cunha, Ana Rita Pedro, João V Cordeiro. Originally published in the Journal of Medical Internet Research (https://www.jmir.org), 10.07.2023.BACKGROUND: The COVID-19 pandemic accelerated the digital transition in health care, which required a rapid adaptation for stakeholders. Telemedicine has emerged as an ideal tool to ensure continuity of care by allowing remote access to specialized medical services. However, its rapid implementation has exacerbated disparities in health care access, especially for the most susceptible populations. OBJECTIVE: We aimed to characterize the determinant factors (facilitators and barriers) of access to hospital medical specialty telemedicine consultations during the COVID-19 pandemic and to identify the main opportunities and challenges (technological, ethical, legal, and social) generated by the use of telemedicine in the context of the COVID-19 pandemic. METHODS: We conducted a systematic review according to PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines. A total of 4 databases (Scopus, Web of Science, PubMed, and Cochrane COVID-19 Study Register) were searched for empirical studies published between January 3, 2020, and December 31, 2021, using established criteria. The protocol of this review was registered and published in PROSPERO (CRD42022302825). A methodological quality assessment was performed, and the results were integrated into a thematic synthesis. The identification of the main opportunities and challenges was done by interpreting and aggregating the thematic synthesis results. RESULTS: Of the 106 studies identified, 9 met the inclusion criteria and the intended quality characteristics. All studies were originally from the United States. The following facilitating factors of telemedicine use were identified: health insurance coverage; prevention of SARS-CoV-2 infection; access to internet services; access to technological devices; better management of work-life balance; and savings in travel costs. We identified the following barriers to telemedicine use: lack of access to internet services; lack of access to technological devices; racial and ethnic disparities; low digital literacy; low income; age; language barriers; health insurance coverage; concerns about data privacy and confidentiality; geographic disparities; and the need for complementary diagnostic tests or the delivery of test results. CONCLUSIONS: The facilitating factors and barriers identified in this systematic review present different opportunities and challenges, including those of a technological nature (access to technological devices and internet services and level of digital literacy), a sociocultural and demographic nature (ethnic and racial disparities, geographic disparities, language barriers, and age), a socioeconomic nature (income level and health insurance coverage), and an ethical and legal nature (data privacy and confidentiality). To expand telemedicine access to hospital-based specialty medical consultations and provide high-quality care to all, including the most susceptible communities, the challenges identified must be thoroughly researched and addressed with informed and dedicated responses.publishersversionpublishe

a panel validation by qualitative Delphi technique

Funding Information: The present publication was funded by Fundação Ciência e Tecnologia, IP national support through CHRC (UIDP/04923/2020). Publisher Copyright: © Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.Background In laboratory medicine, quality and performance indicators (QPIs) are essential tools to ensure the quality of healthcare services and patient safety. QPIs allow comparison of outcomes, favouring accountability and transparency. Internationally, there are some QPI evaluation models, but the fact that they are paid limits their dissemination in smaller/poorer laboratories. In Portugal, each laboratory defines its own QPIs, with no uniformity between institutions. The development of a free QPI panel suitable for anatomical pathology laboratories (APLs) would allow for quality assessment and improvement. Objective To develop a consensual and validated QPI panel suitable for Portuguese APLs. Methods The study was developed in two stages. First, a bibliographic review was carried out, selecting the adequate QPIs. Afterwards, these QPIs were evaluated by experts through the Delphi method, where they could also suggest other pertinent QPIs. Results By the end of the Delphi method, there was a consensus on 64 QPIs (31 for 'structure', 30 for 'process' and 3 for 'result'). The consensual QPIs covered all phases of the total test cycle. The lack of specific anatomical pathology QPIs in the bibliography was noticeable. There was greater consensus on 'process' and 'result' QPIs than on 'structure'. This was supported by the bibliography, where the first ones were more valued. Nevertheless, it is important to monitor all the main laboratory processes, prioritising the evaluation of QPIs with greater impact on healthcare quality and patient safety. These results should allow APLs to identify the causes behind poor performance and improve their services. Conclusions This panel is a valuable tool for APLs, contributing to quality awareness. It can be the first step towards the development of a free benchmarking quality programme in Portugal, encouraging competitiveness and cost-efficiency.publishersversionpublishe

Literacia em saúde: da gestão da informação à decisão inteligente

RESUMO - Introdução: A noção de um sistema de saúde centrado no cidadão e o papel que este pode assumir no controlo e gestão da sua própria saúde têm estado presentes na teoria e prática da saúde pública ao longo dos últimos anos. A centralidade no cidadão é especialmente notória quando acompanhamos a evolução de dois dos pilares da Saúde Pública, a promoção de Literacia em Saúde, enquadrada nos modelos teóricos da educação para a saúde, e a Gestão de Informação de Saúde, apoiada fortemente na evolução dos Sistemas de Informação em Saúde. Objetivos: Analisou-se o momento em que a evolução dos dois pilares convergem, permitindo que estas novas ferramentas de gestão da informação e comunicação se constituíssem instrumentos de promoção da literacia em saúde do cidadão. Assim pretendeu-se (i) traduzir, adaptar culturalmente e validar o European Health Literacy Survey (HLS-EU) para o contexto português (HLS-EU-PT), por forma a poder (ii) avaliar o nível de literacia em saúde da população portuguesa, e subsequentemente comparar com a população europeia (iii) descrever e analisar a decisão partilhada e os sistemas de informação de saúde centrados no cidadão como mediadores de promoção de literacia em saúde, (iv) identificar quais as principais barreiras e fatores facilitadores para a utilização destes mediadores; e (v) descrever e validar uma proposta de Estratégia Nacional de Literacia em Saúde. Métodos: Para a concretização dos objetivos foi seguido um modelo misto com recurso a metodologias quantitativas e qualitativas de recolha e análise de informação. Para tal, foi recolhida informação através de questionário, revisão da literatura, entrevistas, observação participante, análise documental, grupo focal e técnica de grupo nominal. Resultados: O HLS-EU-PT evidencia propriedades psicométricas comparáveis às versões utilizadas nos outros países. Em Portugal, 61,4% da população inquirida apresenta um nível de literacia em saúde problemático ou inadequado. No subíndice Cuidados de Saúde a percentagem é de 55,1%; no Prevenção da Doença 55,1% e no Promoção de Saúde a percentagem sobe para 60,2%. Os sistemas de informação de saúde centrados no cidadão têm evoluído de repositórios estáticos de informação de saúde para instrumentos de apoio ao conhecimento, à literacia em saúde e à negociação de objetivos. A baixa literacia em saúde do cidadão e a falta de consciência da sua utilidade são apontadas como as principais barreiras à utilização de PHIS no contexto de saúde português. Do ponto de vista tecnológico, a falta de acesso ou de suporte tecnológico e a baixa literacia digital são as principais iv barreiras à sua utilização. Do ponto de vista da relação e de sistema acrescem as atitudes negativas do médico e o pouco tempo disponível na consulta. A formação e acompanhamento do cidadão no que diz respeito à sua utilização, a perceção dos benefícios e o envolvimento e apoio do médico no processo são os principais fatores que podem facilitar a sua implementação e utilização em Portugal. Analisando a participação do cidadão nas decisões sobre a sua própria saúde, continua a existir uma tendência para a adoção de um modelo paternalista, no qual o médico se substitui ao cidadão na decisão. O nível de literacia em saúde do cidadão surge como o aspeto que mais dificulta a adoção do modelo de decisão partilhada. Identificam-se também como barreiras o facto de o doente não querer participar, precisar de ser guiado, o seu estado emocional, a doença não permitir, a sua capacidade para comunicar não possibilitar e o não querer assumir responsabilidade. Relacionados com o médico, a suas competências comunicacionais são apontadas como o fator que mais influencia a adoção do modelo partilhado, e com o contexto o pouco tempo disponível para cada consulta. Conclusões: Tendo em conta que os cuidados de saúde centrados no cidadão devem contemplar a promoção da literacia em saúde, da facilitação do acesso e gestão informação de saúde pelo cidadão e da decisão partilhada, conclui-se que, apesar de existirem várias iniciativas a decorrer neste sentido, há ainda um longo caminho a percorrer. A análise dos resultados do HLS-EU-PT revelam níveis de literacia em saúde baixos e particularmente preocupantes nas populações tidas como vulneráveis. Os dois mediadores analisados podem ter um papel fundamental na promoção de literacia em saúde. Mas se por um lado a promovem, por outro a sua utilização é fortemente influenciada por esta, pelo que é fundamental quebrar este circulo. O envolvimento efetivo dos médicos, a formação e acompanhamento do cidadão e a formação dos médicos no que diz respeito às suas competências relacionais e comunicacionais podem ser o caminho para o fazer. A problemática da literacia em saúde não é um problema individual, é sistémica e carece de uma abordagem holística, pelo que a concretização de uma Estratégia Nacional de Literacia em Saúde, deve ser feita em cinco eixos estratégicos: incluir a literacia em saúde no próprio sistema; assegurar o acesso e gestão da informação; assegurar uma comunicação efetiva; integrar a literacia em saúde na educação; e garantir a sustentabilidade das iniciativas.ABSTRACT - Introduction: The conception of a citizen-centred health system and the role that citizens can play in controlling and managing their own health have been present in public health theory and practice over the past few years. The centrality of the citizen is especially evident when we follow the evolution of two pillars of Public Health, (i) the promotion of Health Literacy, framed in the theoretical models of health education, and (ii) Health Information Management, strongly supported in the evolution of citizen-centred health information systems. Aims: This thesis aims to analyse the moment where these two pillars converge, allowing new tools of information and communication management to be powerful instruments in the promotion of citizens' health literacy. The specific aims were (i) to translate, culturally adapt and validate the European Health Literacy Survey (HLS-EU) into the Portuguese context (HLS-EU-PT), in order to be able (ii) to evaluate the level of health literacy of the Portuguese population, and subsequently to compare it with the European population, (iii) to identify, describe and analyse shared decision and personaliwed health information systems as mediators of health literacy promotion, and (iv) to describe and validate a National Health Literacy Strategy proposal. Methods: A mixed model was followed, using quantitative and qualitative approaches to collect and analyse information. The information was collected through questionnaires, literature review, interviews, participant observation, documental analyses, focus groups and nominal group techniques. Results: The HLS-EU-PT shows psychometric properties comparable to the versions used in the other countries. In Portugal, 61.4% of the surveyed population presents a problematic or inadequate level of general health literacy. In the sub-index Health Care the percentage is 55.1%; in Disease Prevention 55.1% and in Health Promotion the percentage rises to 60.2%. Citizen-centered health information systems have evolved from static repositories of personal health information to tools that support knowledge, health literacy, and health goals negotiation. Citizens’ low health literacy and lack of awareness of its usefulness are pointed out as the main barriers to the use of PHIS in the Portuguese health context. From a technological point of view, lack of access or technological support and low digital literacy are the main barriers to its use. From the point of view of the relationship and the system, it adds to the negative attitudes of the doctor and the short time available in the consultation. Citizens’ training and support, as well as their perception of the benefits of vi use that and the doctors’ involvement in the process are the main factors that could enable its implementation and use in Portugal. When analysing the participation of citizens in health decisions, there continues to be a tendency towards the adoption of a paternalistic model, in which the physician substitutes the citizen in the decision. The level of citizenship health literacy appears as the aspect that makes it more difficult to adopt a shared decision model. Also identified as a barrier is the fact that the patient does not want to participate, or needs to be guided, their emotional state, their disease or ability to communicate do not enable to participate and they do not want to take responsibility. As for physicians, their communicational skills are singled out as the factor that most influences the adoption of a shared decision model. Added to these factors is the limited time available in each consultation. Conclusions: Taking into account that citizen-centred health care should include health literacy promotion, citizens’ access and management to their own health information and shared decision, it is concluded that, although there are several initiatives to be taken in this direction, there is still a long way to go. The analysis of HLS-EU-PT results reveals that Portuguese health literacy levels are low and are especially worrying in the populations considered as vulnerable. The two mediators analysed can play a key role in promoting health literacy. However, on the one hand they promote it, on the other hand its adoption and use are strongly influenced by health literacy, so it is fundamental to break this circle. The effective involvement of physicians, the training and support of the citizen and the training of physicians regarding their relational and communicational skills may be the way forward. Taking into account that health literacy is not an individual problem but rather a systemic one, hence needinga holistic approach the implementation of a National Strategy for Health Literacy is suggested, focusing on five strategic axes: include health literacy in the system itself; ensure access to, and management of health information; ensure effective communication; integrate health literacy into education; and ensure the sustainability of initiatives to promote health literacy