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Zika virus became a major public health problem in early 2015, when cases of Guillain–Barré syndrome and microcephaly were associated with viral infection. Currently, ZIKV is endemic in all tropical areas of the world, and the chance for future Zika epidemics remains very real and accurate diagnosis is crucial. The aim of this work was to select specific ssDNA aptamers that bind to the entire Zika virus and can be used to compose specific diagnostics, without cross-reactivity with other flaviviruses. Zika virus was cultivated in Vero cells and used as a target for aptamer selection. Aptamers specific for the ZIKV were selected using whole-virus SELEX, with counterselection for other flavivirus. Secondary and tertiary structures were evaluated and the molecular anchoring between the aptamers and target were simulated by the HDOCK server. Aptamer interaction was evaluated by ELISA/ELASA and the dissociation constant (Kd) was calculated by thermophoresis. Four ZIKV-specific aptamers were selected. The best two were further characterized and proved to be specific for ZIKV. Aptamers are capable of binding specifically to the ZIKV and differentiate from Dengue virus. The aptamers selected in this work can be used as capture agents in the composition of diagnostic tests to specifically detect ZIKV infection.

Within the context of global climate change and overfishing of fish stocks, there is some evidence that cephalopod populations are benefiting from this changing setting. These invertebrates show enhanced phenotypic flexibility and are found from polar regions to the tropics. Yet, the global patterns of species richness in coastal cephalopods are not known. Here, among the 370 identified-species, 164 are octopuses, 96 are cuttlefishes, 54 are bobtails and bottletails, 48 are inshore squids and 8 are pygmy squids. The most diverse ocean is the Pacific (with 213 cephalopod species), followed by the Indian (146 species) and Atlantic (95 species). The least diverse are the Southern (15 species) and the Arctic (12 species) Oceans. Endemism is higher in the Southern Ocean (87%) and lower in the Arctic (25%), which reflects the younger age and the “Atlantification” of the latter. The former is associated with an old lineage of octopuses that diverged around 33 Ma. Within the 232 ecoregions considered, the highest values of octopus and cuttlefish richness are observed in the Central Kuroshio Current ecoregion (with a total of 64 species), followed by the East China Sea (59 species). This pattern suggests dispersal in the Central Indo-Pacific (CIP) associated with the highly productive Oyashio/Kuroshio current system. In contrast, inshore squid hotspots are found within the CIP, namely in the Sunda Shelf province, which may be linked to the occurrence of an ancient intermittent biogeographic barrier: a land bridge formed during the Pleistocene which severely restricted water flow between the Pacific and Indian Oceans, thereby facilitating squid fauna differentiation. Another marked pattern is a longitudinal richness cline from the Central (CIP) towards the Eastern Indo-Pacific (EIP) realm, with central Pacific archipelagos as evolutionary dead ends. In the Atlantic Ocean, closure of the Atrato Seaway (at the Isthmus of Panama) and Straits of Gibraltar (Mediterranean Sea) are historical processes that may explain the contemporary Caribbean octopus richness and Mediterranean sepiolid endemism, respectively. Last, we discuss how the life cycles and strategies of cephalopods may allow them to adapt quickly to future climate change and extend the borealization of their distribution.

Subgenomic RNAs (sgRNAs) are potential markers of active SARS-CoV-2 replication, serving as templates for the synthesis of structural and accessory proteins in infectious viral particles. This study aimed to use RT-qPCR to quantify sgRNA and negative RNA intermediates, assessing viral replication in virus samples inactivated by β-propiolactone (βPL). Inactivated viruses subjected to five blind serial passages (BSs) were amplified by RT-qPCR using primers to target the envelope (ENV) and nucleoproteins (N1 and N2) of genomic genes, subgenomic envelope RNA (sgENV), and intermediate envelope RNA (ENV-). All positive controls showed consistent viral titers across passages (10 log10 copies/mL in N1/N2 and 11 log10 copies/mL in ENV) during BSs. Inactivated viral samples for ENV and ENV- targets ranged from 11.34 log10 copies/mL in BS1 to 11.20 log10 copies/mL in BS5. The sgENV was no longer detected in the inactivated SARS-CoV-2 samples after the second passage, suggesting successful inactivation. Replication kinetics showed consistent profiles for N1/N2, ENV, and ENV- targets in the first three post-infection hours (pih) and maintained approximately 5 log10 copies/mL at 1 pih, 2 pih, and 3 pih. A sharp exponential increase in the viral titer was observed from 24 pih onwards, peaking at 11.64 log10 copies/mL at 48 pih. Transmission electron microscopy confirmed viral particles only in cells infected with active SARS-CoV-2. These results support the use of sgRNA as a reliable marker for SARS-CoV-2 replication, especially in distinguishing between active replication and non-viable particles and in the development of diagnostic and therapeutic strategies.

The most immediate and direct impact of global warming on human health is recognised by the consistent increase in the average global temperature and increase frequency, intensity, and heatwaves duration. This example of extreme heat events is already considered one of the most important climatic hazards and constitute a leading cause of mortality worldwide, particularly in southern Europe with pronounced impacts on cities. The respective health and economic burdens are of growing concern and have attracted various authorities and institutions to find new public health measures to protect citizens.Evidence supports that heat hazard related health outcomes are associated with the populations' exposure, vulnerability, and adaptive capacity. The literature identifies individual risk factors such as age (individuals over 65) and people with pre-existing cardiovascular and/or respiratory diseases. In addition to health and demographic factors, the community's vulnerability is also associated with low socioeconomic conditions, lack of urban vegetation, low education attainment, building characteristics, among others. Less studied are the population adaptation and adaptive capacity.Portugal has a well-documented heat-related morbidity and mortality history. It has a set of well-established models for heat-related mortality either for the city of Lisbon and for Portugal mainland (joining models for four ad-hoc constructed geographical regions) – known as ÍCARUS' models. Based on these models Portugal has a well-established Heat Health Warning System (HHWS) since 1999 – the ICARUS Surveillance System, recognised in scientific literature as the first of such systems in Europe.The studies present in this thesis aimed: 1) to analyse the eventual need to update the existing ICARUS′ models and surveillance system, 2) and to investigate the potential advantage of studying mortality at the neighbourhood scale of Lisbon's municipality, the respective spatial patterns of heat-related mortality and their potential explanatory factors. The performed work building from the evolution in Geographic Information Systems (GIS) and the currently associated sets of analytical and statistical methods went beyond conventional geographical scales analysis to align with the real people's needs and improve health outcomes' spatial patterns knowledge and accuracy.This dissertation's work showed that the national models for heat-related mortality, particularly the Lisbon model, and inherently the existing Surveillance System, do not need updating. The heat-related mortality metric for Lisbon models GATO IV (Generalized Accumulated Thermal Overload) has a good predictive power to prevent health heat-related risks performing better for Lisbon than the international metric EHF (Excess Heat Factor), more broadly known, cited, and referenced in the scientific community. It was also showed how to geocode deaths in Portugal using the addresses present on e-death certificate and, subsequently, its mapping elderly's heat-related cardiorespiratory mortality at a neighbourhood scale for Lisbon. It was possible to understand the potential spatially-varying factors, from medical facilities to socioeconomic, urbanistic and environmental factors also associated with each neighbourhood.To our knowledge, this is the first spatial modelling of heat-related mortality at this scale, and with such a wide range of variables.

ABSTRACT Introduction: The emergence of orthodontic aligners has provided an aesthetic and comfortable option for orthodontic treatment. However, the encapsulated design of the aligners can influence the masticatory muscles, and might compromise safe treatment. Objective: This preliminary longitudinal study aimed to investigate whether the use of orthodontic aligners affects the biting force and myoelectric activity of the superficial masseter and anterior temporal muscles. Methods: Ten subjects participated in the study and underwent treatment during an 8-month follow-up period. The root mean square (RMS), the median power frequency (MPF) of the surface electromyography (sEMG) signals, and the biting force (kgf) were recorded and normalized relative to the pretreatment condition. The data were analyzed by repeated-measure analysis of variance (ANOVA), with the significance level set at 5%. Results: Both the superficial masseter and the anterior temporal muscles presented an increase in sEMG signal activity during the treatment, with a marked increase in the latter compared to the former (p<0.05). Moreover, a significant decrease in bite force was evidenced (p<0.05). Conclusions: This preliminary study observed that the orthodontic aligners affected the muscle recruitment pattern of masticatory muscles, and reduced biting performance during the 8-month follow-up period. RESUMO Introdução: O surgimento dos alinhadores ortodônticos tem proporcionado uma opção estética e confortável para o tratamento ortodôntico. No entanto, o design encapsulado dos alinhadores pode influenciar os músculos mastigatórios e comprometer a segurança do tratamento. Objetivo: Este estudo longitudinal preliminar teve como objetivo investigar se o uso de alinhadores ortodônticos afeta a força de mordida e a atividade mioelétrica dos músculos masseter superficial e temporal anterior. Métodos: Dez indivíduos participaram do estudo e foram submetidos a tratamento durante um período de acompanhamento de 8 meses. A raiz quadrada média (RMS), a frequência mediana de potência (FMP) dos sinais de superfície da eletromiografia (sEMG) e a força de mordida (kgf) foram registradas e normalizadas em relação à condição de pré-tratamento. Os dados foram analisados por análise de variância para medidas repetidas (ANOVA), com nível de significância estabelecido em 5%. Resultados: Tanto o masseter superficial quanto o temporal anterior apresentaram aumento da atividade do sinal sEMG durante o tratamento, com aumento acentuado desse último em comparação ao primeiro (p<0,05). Além disso, foi evidenciada uma diminuição significativa da força de mordida (p<0,05). Conclusões: Esse estudo preliminar observou que os alinhadores ortodônticos afetaram o padrão de recrutamento muscular dos músculos mastigatórios e reduziram o desempenho da mordida durante o período de acompanhamento de oito meses.

Background: COVID-19 is caused by SARS-CoV-2 infection and has reached pandemic proportions. Since then, several clinical characteristics have been associated with poor outcomes. This study aimed to describe the morbidity profile of COVID-19 deaths in Portugal. Methods: A study was performed including deaths certificated in Portugal with “COVID-19” (ICD-10: U07.1 or U07.2) coded as the underlying cause of death from the National e-Death Certificates Information System between 16 March and 31 December 2020. Comorbidities were derived from ICD-10 codes using the Charlson and Elixhauser indexes. The resident Portuguese population estimates for 2020 were used. Results: The study included 6701 deaths (death rate: 65.1 deaths/100,000 inhabitants), predominantly males (72.1). The male-to-female mortality ratio was 1.1. The male-to-female mortality rate ratio was 1.2; however, within age groups, it varied 5.0–11.4-fold. COVID-19 deaths in Portugal during 2020 occurred mainly in individuals aged 80 years or older, predominantly in public healthcare institutions. Uncomplicated hypertension, uncomplicated diabetes mellitus, congestive heart failure, renal failure, cardiac arrhythmias, dementia, and cerebrovascular disease were observed among COVID-19 deceased patients, with prevalences higher than 10%. A high prevalence of zero morbidities was registered using both the Elixhauser and Charlson comorbidities lists (above 40.2%). Nevertheless, high multimorbidity was also identified at the time of COVID-19 death (about 36.5%). Higher multimorbidity levels were observed in men, increasing with age up to 80 years old. Zero-morbidity prevalence and high multimorbidity prevalences varied throughout the year 2020, seemingly more elevated in the mortality waves’ peaks, suggesting variation according to the degree of disease incidence at a given period. Conclusions: This study provides detailed sociodemographic and clinical information on all certificated deaths from COVID-19 in Portugal during 2020, showing complex and extreme levels of morbidity (zero-morbidity vs. high multimorbidity) dynamics during the first year of the pandemic in Portugal.

Objetivo–O objetivo deste estudo é analisar a importância da internacionalização da indústria criativa em Portugal e Brasil para o progresso interno e identificar políticas públicas eficientes que possam estimular o desenvolvimento e a internacionalização dessa indústria e assim, promover maior evolução sócio-econômico-cultural das populações.Metodologia–Foi realizada uma pesquisa descritiva de levantamento bibliográfico e informações publicadas pelos órgãos de governo responsáveis pela mensuração do impacto do mercado empresarial nos países.Resultados e conclusões–Ficou claro o impacto positivo dessas empresas tanto em Portugal, quanto no Brasil e o quanto vem crescendo sua importância em cada um dos países tanto internamente, quanto no exterior. Também foi apresentada o histórico de políticas públicas voltados para o setor nos dois países e realizada uma série de propostas de políticas públicas com o objetivo de contribuir com os organismos governamentais e com o desenvolvimento das empresas e das cidades onde essa indústria escolhe se estabelecer.Limitações e implicações–Uma das limitações apontadas refere-se ao fato de que são recentes as pesquisas e o mapeamento pelos países sobre o setor criativo. Outro ponto limitante relaciona-se à diversidade de tipos de empresas que compõem a indústria criativa e que vivem desafios bem diferentes quando se internacionalizam. Isso faz com que seja complexo estabelecer um mecanismo de ajuda que seja efetivo para todo o setor.Originalidade e valor–Este estudo teve o intuito de reunir informações e contribuir com material que embase a criação de políticas públicas para estimular as indústrias criativas, principalmente de Portugal e do Brasil, a se desenvolverem e se internacionalizarem. E com isso, aumentar o desenvolvimento sócio-econômico do país de origem e o de destino e aumentar o destaque e prestígio das nações no mercado internacional. Também tem seu valor por ter relacionado a internacionalização do setor criativo com o desenvolvimento interno dos países

As vagas de calor ou os dias de calor excessivo, têm vindo a ocorrer nos últimos decénios com maior frequência, intensidade e extensão geográfica, um pouco por todo o mundo. Associam-se a estas anomalias térmicas as consequências na saúde pública e mesmo o aumento da mortalidade diária, com especial incidência nas aglomerações urbanas.O principal objectivo desta dissertação é identificar, na cidade de Lisboa, as áreas de maior perigo às temperaturas elevadas. Para o efeito foi desenvolvido um modelo espacial da probabilidade de óbito associado ao calor extremo, considerando a relação perigosidade/vulnerabilidade, para estimar as dissimetrias espaciais na cidade de Lisboa. Tratase de uma análise integrada, operando no modelo variáveis climáticas (como a temperatura do ar e a energia disponível na superfície obtida através de uma imagem de Satélite Landsat 7 ETM+) e outras de natureza socioeconómica e indicadores urbanísticos (taxa de desemprego, taxa de analfabetismo, idade dos edifícios, área verde privada, entre outros). O estudo baseia-se em regressões lineares múltiplas entre a mortalidade e os diversos indicadores.Com o cruzamento das duas componentes (perigosidade/vulnerabilidade) obteve-se a susceptibilidade da ocorrência de óbito devido às temperaturas excessivas. Verificou-se que a variação espacial da mortalidade é essencialmente explicada pelas variáveis correspondentes à componente da vulnerabilidade, detendo o modelo uma capacidade explicativa de 68%. Em linhas gerais, a probabilidade de óbito é mais elevada nas áreas com piores condições socioeconómicas, nomeadamente onde é elevada a taxa de desemprego e a taxa de analfabetismo, onde existe menos espaço verde privado e árvores de arruamento e menos população imigrante. A distinção das áreas de maior susceptibilidade é disso exemplificativo. O núcleo central de Lisboa é mais susceptível à ocorrência de óbito que a área norte e ocidental da cidade, menos vulnerável às temperaturas extremas.Esta análise pretende ser uma base de discussão e um ponto de partida para uma melhor modelação ao calor, ao mesmo tempo que evidencia o contributo da Detecção Remota e dos Sistemas de Informação Geográfica na espacialização dos fenómenos climáticos extremos e suas consequências, como é a susceptibilidade a temperaturas muito levadas na cidade de Lisboa.

Heavy metals such as nickel (Ni) can lead to bioaccumulation, affecting entire ecosystems and posing significant risks to various life forms, including plants. Although Ni is a micronutrient, it can be toxic by impacting enzyme activities and inhibiting seed germination and plant growth. In Brazil, official guidelines stipulate preventive and intervention values for Ni concentrations in soil to mitigate pollution and protect soil and groundwater quality. Soil samples classified as Typic Haplustox were artificially contaminated with NiCl₂ at concentrations of 120 (T120), 240 (T240), and 360 (T360) mg dm-3, alongside a control treatment (T0). Seeds were cultivated under greenhouse conditions, and germination and growth parameters were analyzed after 15 days. Measurements included emergence speed index, germination percentage, root and shoot length, fresh and dry mass, and biochemical analyses of nitrogenous compounds and sugars. This study addresses the effects of toxic concentrations of NiCl2 on plants, focusing on the germination and early growth stages. Canavalia ensiformis (L.) DC., a tropical legume with significant roles in green manure and phytoremediation, was chosen for its adaptability to various soils. The hypothesis is that C. ensiformis can withstand high soil Ni concentrations, maintaining growth despite environmental toxicity limits. The results indicated differential impacts of Ni, the emergence percentage decreases at 360 mg dm-3 soil with greater dry mass accumulation at 120 and 240 mg dm-3, highlighting the importance of understanding plant responses to stress from potentially toxic elements for sustainable agricultural practices and environmental management.

Background Communication skills are essential for health professionals to establish a positive relationship with their patients, improving their health and quality of life. In this perspective, communication skills training can be effective strategies to improve the care provided by professionals in patient care and the quality of health services. Objective To identify the best available evidence on training programs in communication skills to promote changes in attitude and behavior or self-efficacy of health professionals. Methods Systematic searches were performed in eight databases, evaluating Randomized Controlled Trials and quasi-experimental studies with a control group, focusing on training communication skills for health professionals, who assessed self-efficacy or behaviors related to these skills. The phases of study selection and data extraction were carried out by two independent researchers, and the conflicts were resolved by a third. The risk of bias was assessed using the Cochrane method. Results Eight studies were included in the review. Most programs lasted between 4½ h and 2 days, involved information about communication skills and the content was applied to the health professionals’ context. Several teaching strategies were used, such as lectures, videos and dramatizations and the evaluation was carried out using different instruments. Improvements in the performance and in the self-efficacy of communication skills were observed in the trained groups. The RCT had a low risk of bias and the quasi-experimental studies had a moderate risk. Conclusion Training in communication skills can improve the performance and self-efficacy of health professionals. Programs that approach the conceptual issues and promote the space for experiential learning could be effective in communication skills training for professionals. PROSPERO : CRD42019129384