National Institute of Health Dr. Ricardo Jorge
Repositório Científico do Instituto Nacional de SaúdeNot a member yet
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Identification of mTOR and AGO1 IRES trans-acting factors
No full textCancer is the second leading cause of death globally; therefore, its study is crucial to discover new therapies. Under stress, the regular process of protein synthesis (canonical translation) is impaired, while a back-up mechanism mediated by internal ribosome entry sites (IRES) continues to function, allowing the synthesis of proteins that maintain cellular viability. This also happens in cancer cells, contributing for their survival and consequent tumorigenesis. IRES-mediated translation and its regulation by IRES trans-acting factors (ITAFs) has been correlated to metastasis and chemotherapeutic drug resistance. Therefore, our main goal was to validate ITAFs and assess their significance in cancer onset, thus becoming candidates as novel therapeutic targets. A bicistronic reporter system, which contains a first cistron translated via canonical translation and a second one translated by IRES of mTOR1 and AGO12 was used to test IRES-driven translation initiation activity. Experiments were carried out in which several proteins (hnRNPs) were silenced by specific siRNAs to analyse their function as ITAFs of mTOR and AGO1 IRESs. Also, distinct drugs were applied to simulate endoplasmic reticulum (ER) or hypoxia stress, to evaluate their effect on IRES activity. The relative IRES activity was assessed by luminescence tests and the protein levels by Western blot. In general, knockdown of hnRNPK and hnRNPU seems to decrease the IRES activity by ~60% and ~30% respectively, while hnRNPC knockdown does not show a significant effect. Regarding the ER stress, hnRNPK knockdown seems to decrease even more the IRES activity, while hnRNPU depletion induces a significant increase. On the other hand, in hypoxia, the hnRNPs knockdowns do not significantly affect IRES activity. These results indicate that hnRNPK and hnRNPU may function as ITAFs of mTOR and AGO1 IRES activity in cells under ER stress. Our data can be decisive for a better understanding of carcinogenesis and suggest new therapeutic targets for cancer treatment.
1. Marques-Ramos, A., et.al. 2017. RNA. 23, 1712-1728
2. Lacerda, R. 2016. Faculdade de Ciências e Tecnologia da Universidade NOVA de LisboaWork partially supported by UID/MULTI/04046/2019 Research Unit grant from FCT, Portugal (to BioISI)info:eu-repo/semantics/publishedVersio
CCL2 expression predicts clinical outcomes and regulates E-cadherin and angiogenesis in pituitary tumours
No full textThis paper is part of a special collection highlighting the work of emerging leaders in the endocrine cancer field.The crosstalk between tumour cells and microenvironment components in pituitary neuroendocrine tumours (PitNETs), including chemokines, may impact tumour behaviour and clinical outcomes. CCL2 was previously identified as a key chemokine in PitNETs, but its role remains unknown. We aimed to study the role of CCL2 in defining the phenotype and clinical outcomes of PitNETs and in regulating macrophage chemotaxis, epithelial-to-mesenchymal transition (EMT) and angiogenesis. We studied CCL2 and E-cadherin expression, macrophages (CD68 and CD163) and vessels (CD31) in samples from 86 PitNET patients. Higher CCL2 mRNA expression was found in patients who required multimodal and multiple treatments and had active disease at the last follow-up. Higher CCL2 immunoreactivity was observed in patients with larger PitNETs. Among somatotroph tumours, CCL2 mRNA expression correlated with serum IGF-1 at the last follow-up. CCL2 mRNA expression levels correlated negatively with CDH1 expression and with E-cadherin complete membranous staining. In vitro, CCL2 downregulated E-cadherin expression in GH3 cells but did not affect cell morphology or migration. CCL2 expression correlated with the number of vessels, vessel perimeter and vessel area in PitNETs but not with PitNET-infiltrating macrophages. Our data suggest that CCL2 may lead to (or is at least a predictive marker of) poorer clinical outcomes and more difficult-to-treat PitNETs, potentially through its regulatory effects on different tumour-related mechanisms beyond immune cell chemotaxis, including in the activation of the EMT pathway and modulation of angiogenesis in PitNETs. Further studies are needed to corroborate our findings and to validate CCL2 as a potential predictive marker and therapeutic target in PitNETs.PM was supported by a Pilot Project Award from the Neuroendocrine Tumor research Foundation (NETRF) and by the Maria de Sousa Prize awarded by Fundação BIAL (BIAL Foundation) and Ordem dos Medicos (Portuguese Medical Order)
Scientific opinion on the safety of a proposed amendment of the conditions of use of the food additive sorbitan monostearate (E 491) in enzyme preparations
No full textThe EFSA Panel on Food Additives and Flavourings (FAF Panel) provides a scientific opinion on the safety evaluation of a proposed amendment of the conditions of use of the food additive sorbitan monostearate (E 491) in accordance with Annex III, Part 3 to Regulation (EC) No 1333/2008, with respect to the intended use as a food additive in preparations of the food enzyme asparaginase (also known as acrylamide reducing yeast, or ARY). The group of sorbitan esters (E 491–495) was re‐evaluated by the EFSA Panel on Food Additives and Nutrient Sources added to Food (ANS Panel) in 2017. The ANS Panel established a group ADI of 10 mg sorbitan/kg body weight (bw) per day applicable to the food additives E 491–495. In the present opinion the Panel calculated an updated dietary exposure estimate of sorbitan resulting from the current authorised uses of the group of sorbitan esters (E 491–495), and from the proposed amendment of the conditions of use of sorbitan monostearate (E 491) in enzyme preparations. In updating the dietary exposure with the latest dietary surveys available, the group ADI of 10 mg sorbitan/kg bw per day was exceeded in toddlers and children at the 95th percentile in the refined non‐brand loyal scenario for a limited number of dietary surveys. This observation holds true either considering the proposed amendment of the conditions of use of the food additive E 491 or only the currently permitted uses in the exposure calculations. The same conclusions apply to the dietary exposure estimates for consumers of food supplements, for which the ADI is exceeded in children at the 95th percentile. The Panel however concluded that the conservative assumptions made in the refined scenarios have resulted in a clear overestimation of the dietary exposure and therefore that the calculated exceedance of the acceptable daily intake (ADI) is not of safety concern. The Panel concluded that the proposed amendment of the conditions of use of sorbitan monostearate (E 491) in preparations of the food enzyme ARY has little impact on the current dietary exposure to sorbitan resulting from the already permitted uses and reported use levels of sorbitan esters (E 491–495) and would not be of safety concern
Food Choice, Nutrition, and Public Health
No full text(This article belongs to the Special Issue Food Choice, Nutrition, and Public Health)EditorialMaintaining a healthy diet throughout life helps prevent all forms of malnutrition, thereby reducing the risk of non-communicable diseases (NCDs) and related conditions .
Dietary habits, lifestyle choices, and social factors significantly influence our health. Recently, we have seen shifts in food consumption patterns driven by various factors. These include the increased availability of processed foods, rapid urbanization, and changing lifestyles.
The types, amounts, and frequency of consumed foods and beverages define dietary patterns, which have been evolving in recent decades due to the emergence of new or adapted eating habits. Common examples of dietary patterns include the ‘Western dietary pattern’ and the ‘Mediterranean dietary pattern’. Other significant patterns are the ‘prudent dietary pattern’, which emphasizes a high intake of vegetables, fruits, legumes, whole grains, fish, and seafood, and the ‘vegetarian/plant-based dietary pattern’, which entirely omits meat and animal products.
Understanding the individual motives that drive certain food choices is crucial for changing consumption habits, promoting healthier behaviors, and fostering sustainability.
This Editorial introduces the Special Issue “Food Choice, Nutrition, and Public Health” and highlights key topics on this subject
Registo Nacional de Anomalias Congénitas: Regiões 2011-2019
No full textRelatório do Registo Nacional de Anomalias Congénitas – Relatório Regiões, referente aos anos de 2011 e 2019, que apresenta os resultados a nível nacional e desagregados a nível das regiões, utilizando-se a Nomenclatura das Unidades Territoriais para Fins Estatísticos, NUTS II.
No Relatório RENAC−Regiões apresentado, sobre prevalência de anomalias congénitas e fatores associados, destacam-se os seguintes resultados: - As cardiopatias congénitas mantêm-se como o grupo de Anomalias Congénitas (AC) mais prevalente, tanto a nível nacional, como em todas as regiões; - A nível nacional, 58,6% dos casos notificados com AC foram detetados durante a gravidez, variando, esta frequência, entre 71,1% na Região Norte e 34,6% na Região Autónoma da Madeira (RAM); - Tanto a nível nacional, como em todas as regiões, é na idade materna ≥ 40 anos que se observa a maior prevalência de nascimentos com AC; - Na Área Metropolitana de Lisboa (AML) (23,3%) e na Região Algarve (21,3%), observou-se a maior percentagem de grávidas estrangeiras a ter um nascimento com AC. Nas restantes regiões, a frequência variou entre 1,5% (Região Autónoma dos Açores (RAA)) e 7,0% (Região Alentejo); - O consumo de tabaco durante o 1.º trimestre da gravidez foi reportado, mais frequentemente, pelas grávidas da Região Alentejo (14,1%), seguido da RAA (13,5%) e da Região Algarve (13,2%). Nas restantes regiões os consumos variaram entre os 0,8% (RAM) e os 10,1% (AML).
O RENAC, com dados disponíveis desde 1997, é um registo nosológico de base populacional que recebe notificações da ocorrência de Anomalias Congénitas (AC) em Portugal, sendo reportados os casos com AC diagnosticados nos recém-nascidos, fetos-mortos e nos fetos submetidos a interrupção médica da gravidez, após o diagnóstico de malformação grave.
São objetivos deste registo: i) fornecer informação essencial sobre a epidemiologia das AC em Portugal; ii) manter um sistema de vigilância que permita detetar novas exposições teratogénicas; iii) avaliar o impacto do diagnóstico pré-natal; iv) manter uma base de dados disponível para a investigação na área das AC, acessível a profissionais de saúde e à comunidade cientifica; v) integrar a rede europeia de registos de AC
The buccal micronucleus cytome assay: applicability in human biomonitoring
No full textAs exposições ambientais e ocupacionais a alguns agentes químicos ou físicos representam sérios riscos para a saúde humana. A biomonitorização humana (BMH) permite quantificar a exposição a substâncias químicas e seus efeitos precoces através da caracterização de biomarcadores de exposição e de biomarcadores de efeito. O ensaio do micronúcleo em células bucais (MNBMCyt) é um biomarcador precoce de genotoxicidade que permite identificar instabilidade cromossómica em células da mucosa oral, tendo vindo a encontrar uma aplicação crescente em estudos de BMH.
Nesta revisão pretendeu-se dar uma perspetiva da aplicação do MNBMCyt em estudos epidemiológicos realizados em contexto ambiental e ocupacional, com base em exemplos de estudos de BMH recolhidos através de pesquisa bibliográfica.
Globalmente, a facilidade da colheita e processamento da amostra, aliada à já existente harmonização de procedimentos de análise e à sensibilidade para deteção precoce de efeitos genotóxicos decorrentes da exposição a variados agentes químicos (p.ex., compostos orgânicos voláteis, metais, entre outros) e físicos (radiação ionizante) têm levado a propor a sua mais frequente inclusão em futuros estudos de BMH. Contudo, ainda permanecem algumas limitações analíticas (análise microscópica morosa) e necessidade de mais dados que permitam fortalecer a relevância e grau de confiança neste biomarcador, para reforçar a sua futura utilização para apoiar intervenções que visem prevenir o desenvolvimento de doenças relacionadas com a exposição a substâncias genotóxicas.Environmental and occupational exposures to some chemical or physical agents pose serious risks to human health. Human biomonitoring (HBM) al lows the quantificat ion of exposure to chemical substances DSA their early effects through the characterization of exposure and effect biomarkers. The micronucleus (MN) assay in buccal cells (MNBMCyt) is a biomarker of early effect that allows the identification of chromosomal instability in oral mucosa cells, and has been increasingly used in HBM studies.
This review aimed to provide an over view of the application of MNBMCyt in epidemiological studies carried out in environmental and occupational settings, based on examples of HBM studies collected through literature search. Overall, the simplicity of sample collection and processing, combined with the already existing harmonization of analysis procedures and the sensitivity for early detection of genotoxic effects from exposure to various chemical agents (e.g., volatile organic compounds, metals, among others) and physical agents (ionizing radiation) have led to the proposal for its more frequent inclusion in future HBM studies. However, some analytical limitations (time-consuming microscopic analysis) and a need for more data to strengthen the relevance and degree of confidence in this biomarker have been also identified, in order to reinforce its future use to support interventions aimed at preventing the development of diseases related to exposure to genotoxic substances
Lymphogranuloma venereum (LGV) ompA-subvariants of the Portuguese collection of Chlamydia trachomatis, 2007–2023
No full textBackground: Lymphogranuloma venereum (LGV) is a sexually transmitted infection caused by Chlamydia trachomatis ompA-genotypes L1–L3, with increasing numbers of detected cases across Europe. Here, we analysed diversity and temporal distribution of the LGV ompA-subvariants detected in Portugal between 2007 and 2023, in order to better understand the dissemination and diversification landscape of LGV strains.
Methods: The collection of the Portuguese National Reference Laboratory includes 1188 LGV ompA-genotyped samples between 2007 and 2023. In-depth analysis of the diversity of LGV ompA-subvariants circulating in Portugal across the years was performed, identifying newly described subvariants and integrating this data in a comprehensive compilation with all representative LGV ompA-subvariants described globally.
Results: L2 ompA-variant (L2/434/Bu) was consistently the most frequently detected in our collection, with annual proportions ranging from 34.0% to 82.9%, between 2016 and 2023. L2bV5 was the second most frequent followed by L2b, ranging from 5.0% to 27.9% and 2.6% to 23.7% across the years, respectively, from 2017 to 2023. We highlighted the emergence and considerable increase in circulation of L1-like ompA-subvariants in recent years, representing 13.7% of LGV sequences in 2023. We also identified 13 novel LGV ompA-subvariants that had not been described before, differing by up to three mutations from the respective genotype reference sequences.
Conclusions: This study contributes to the worldwide picture of the LGV molecular epidemiology, highlighting the importance of long-term molecular surveillance to monitor the circulation and geographical spread of LGV and to timely identify and track new strains, such as the recently emerging L1-like ompA-subvariants
Rapid climate action is needed: comparing heat vs. COVID-19-related mortality
No full textThe impacts of climate change on human health are often underestimated or perceived to be in a distant future. Here, we present the projected impacts of climate change in the context of COVID-19, a recent human health catastrophe. We compared projected heat mortality with COVID-19 deaths in 38 cities worldwide and found that in half of these cities, heat-related deaths could exceed annual COVID-19 deaths in less than ten years (at + 3.0 °C increase in global warming relative to preindustrial). In seven of these cities, heat mortality could exceed COVID-19 deaths in less than five years. Our results underscore the crucial need for climate action and for the integration of climate change into public health discourse and policy.AMVC acknowledges funding from the Swiss National Science Foundation (TMSGI3_211626). L.S.G. has received funding from the European Union’s Horizon Europe Framework Programme under the Marie Skłodowska-Curie Grant Agreement No. 101064940
Non-polio enterovirus serotypes associated with central nervous system infections in children: evaluation of laboratory cases, 2023–2024
No full textOs enterovírus não-polio permanecem como os principais agentes patogénicos do sistema nervoso central (SNC) em todo o mundo, especialmente em crianças com idade inferior a 5 anos, causando desde doença ligeira a quadros graves que podem evoluir para desfechos fatais.
Entre 2023 e 2024 foram analisadas no INSA fezes de 327 crianças com suspeita de infeção por enterovírus. O diagnóstico laboratorial incluiu a deteção do RNA viral por RT-PCR em tempo real e o isolamento viral. A identificação viral foi realizada por sequenciação genómica.
Neste período foram identificados 109 (33,3%) casos de infeção a enterovírus, dos quais 26 casos (23,9%) corresponderam a infeções do SNC, maioritariamente em crianças com menos de 5 anos (88,5%), sem uma evidente distribuição temporal. Os enterovírus mais frequentes foram os echovírus (tipos 5, 6,7, 9, 18, 21, 25, 30, 31 e 32), detetados em 13 casos (52,0%), seguidos pelos Coxsackievírus A (tipos 4, 6, 9, 10 e 16), em 7 casos (28,0%), ambos associados a meningite e meningoencefalite. O Enterovírus 71 (EV71) foi detetado em dois casos, um de romboencefalite e um de paralisia flácida aguda (PFA), e um caso de meningite esteve associado ao EV-C99. Não se observou nenhum padrão de surto nestes dois anos.
Estes resultados reforçam a relevância da identificação rápida e dirigida para enterovírus como agentes etiológicos de infeções do SNC, fundamental para uma intervenção clínica direcionada, evitando o uso desnecessário de antibióticos e de múltiplos exames complementares de diagnósticoNon-polio enteroviruses remain the major pathogens of the central nervous system (CNS) worldwide, especially in children under five years of age causing a range of diseases from mild to severe, which can lead to fatal outcomes.
Between 2023 and 2024, stool samples from 327 children with suspected enterovirus infection were analysed at INSA. Laboratory diagnosis included detection of viral RNA by real-time RT-PCR and viral isolation. Serotype identification was performed through genomic sequencing.
During this period, 109 (33.3%) cases of enterovirus infection were identified, of which 26 (23.9%) involved CNS infections, predominantly in children under 5 years of age (88.5%), without an evident temporal distribution. The most frequent enteroviruses were echoviruses (types 5,6, 7, 9, 18, 21, 25, 30, 31, and 32), detected in 13 cases (52.0%), followed by Coxsackievi ruses A (types 4, 6, 9, 10, and 16), in 7 cases (28.0%), both associated with meningitis and meningoencephalitis. Enterovirus 71 (EV71) was identified in two cases, one of rhombencephalitis and one of acute flaccid paralysis (AFP), while EV-C99 was associated with a case of meningi tis. No outbreak pattern was observed.
These findings underscore the importance of rapid and targeted identification of enteroviruses as causative agents of CNS infections, which is crucial for guiding appropriate clinical management and preventing unnecessary use of antibiotics and multiple complementary diagnostic procedures
