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- Relatório REVIVE 2024 - Culicídeos, Ixodídeos e Flebótomos: Rede de Vigilância de VetoresPublication . Centro de Estudos de Vetores e Doenças Infeciosas Doutor Francisco CambournacRelatório REVIVE - Rede de Vigilância de Vetores relativo às atividades desenvolvidas em 2024, que apresenta os resultados da vigilância de culicídeos, ixodídeos e flebótomos. O programa REVIVE visa monitorizar a atividade de artrópodes hematófagos, caracterizar as espécies e sua ocorrência sazonal, e identificar agentes patogénicos importantes em saúde pública. Das atividades apresentadas no presente relatório, destaca-se o seguinte: REVIVE – Culicídeos: - Participaram as cinco Regiões de Saúde e a Direção Regional de Saúde da Madeira, entidades que realizaram colheitas de mosquitos em 241 concelhos de Portugal; - No total foram identificados 38522 mosquitos de 18 espécies, assim como 65802 ovos de espécies invasoras. Nas amostras em que foi pesquisada a presença de flavivírus e alfavírus patogénicos para o Homem, os resultados foram negativos; - O mosquito invasor Aedes aegypti está presente na Região Autónoma da Madeira desde 2005. Outra espécie de mosquitos invasor, Aedes albopictus, foi identificado, pela primeira vez, na região Norte de Portugal em 2017, no Algarve em 2018, no Alentejo em 2022, na região de Lisboa em 2023 e na região Centro em 2024. Estas espécies são vetoras de vírus como dengue, Zika e chikungunya, e têm vindo a aumentar a sua distribuição geográfica nestas regiões. Aedes albopictus foi identificado em 20 concelhos do país em 2024; - No âmbito do REVIVE – Culicídeos foi feita a vigilância em cinco aeroportos internacionais, dois aeródromos, catorze portos e dez outros pontos de entrada de acordo com o Regulamento Sanitário Internacional. REVIVE – Ixodídeos - Participaram as cinco Regiões de Saúde e a Direção Regional de Saúde da Madeira, entidades que realizaram colheitas de carraças em 218 concelhos; - No total foram identificadas 4797 carraças. Para além de 12 espécies de Ixodidae já anteriormente identificadas em Portugal, foram identificados exemplares exóticos de Argasidae que foram classificados como Argas e Ornithodoros spp.; - Nas 1378 carraças utilizadas na pesquisa de Borreliae Rickettsia, em 6,7% e 22,7% respetivamente, foi detetado DNA destes agentes. Nem todas as espécies detetadas são patogénicas para o Homem. - Em 2024 destaca-se assim a deteção de quatro espécies de Borreliae quatro espécies de Rickettsia, já associadas a casos de doença no Homem em Portugal: afzelii, B. garinii, B. lusitaniae e B. valaisiana, agentes etiológicos da borreliose de Lyme; R. conorii agente da febre escaro-nodular; R. monacensis e R. raoultii sem denominação da doença e R. slovaca, agente responsável de TIBOLA. A pesquisa do vírus da febre hemorrágica Crimeia-Congo (CCHFV) foi realizada em 124 exemplares de carraças do género Hyalomma com resultados negativos. REVIVE – Flebótomos - Participaram as cinco Regiões de Saúde, com colheitas realizadas em 50 concelhos; - No total foram colhidos 1046 flebótomos, tendo sido identificados exemplares pertencentes às espécies Phlebotomus ariasi, Ph. perniciosus, Ph. Sergente e Sergentomyia minuta; - Nos 616 flebótomos pesquisados para a presença de flebovírus e de Leismania spp. foi detetada a presença de Leishmania infantum num pool.
- Transcriptomic analysis and epigenetic regulators in human oocytes at different stages of oocyte meiotic maturationPublication . Caniçais, Carla; Sobral, Daniel; Vasconcelos, Sara; Cunha, Mariana; Pinto, Alice; Guimarães, Joana Mesquita; Santos, Fátima; Barros, Alberto; Dória, Sofia; Marques, C. JoanaHuman oocytes are highly specialized cells with the capacity to store and regulate mRNAs during oocyte maturation, in preparation for post-fertilization steps. Here we performed single-oocyte transcriptomic analysis of human oocytes in three meitoic maturation stages - Germinal Vesicle (GV; n = 6), Metaphase I (MI; n = 6) and Metaphase II (MII; n = 7). Single-oocyte transcriptomic analysis revealed that the total number of expressed genes progressively decreased from GV to MII stages, with 9660 genes being transcribed in GV, 8734 in MI and 5889 in MII. The same tendency was observed for the number of uniquely expressed genes, with 1328 uniquely expressed genes in GV, 401 in MI and 72 in MII. GO analysis of the uniquely expressed genes showed distinct terms in GV oocytes such as transferase activity, organonitrogen compound metabolic process and ncRNA processing. Analysis of Differentially Expressed Genes (DEGs) between the three maturation stages revealed 1165 DEGs between GV and MII oocytes, with 635 being upregulated and 528 downregulated, 42 DEGs between GV and MI, with 38 being upregulated and 4 downregulated, and no significant changes in gene expression between MI and MII oocytes. Comprehensive analysis of epigenetic regulators showed high expression of several histone-modifying enzymes, namely deacetylases, acetylases, lysine demethylases and methyltransferases, and DNA methylation regulators, namely the maintenance methyltransferase DNMT1 and its co-regulators DPPA3 and UHRF1. Some of these epigenetic regulators were differentially expressed between maturation stages, namely SIRT3, SIRT6, KDM3AP1, KMT2E, DNMT1, DPPA3 and the MEST and RASGRF1 imprinted genes. Our study contributes with important information on the transcriptional landscape of human oocytes in different stages of meiotic maturation, providing important insights into candidate biomarkers of human oocyte quality.
- Red blood cell proteomic profiling in mild and severe obstructive sleep apnea patients before and after positive airway pressure treatmentPublication . Valentim-Coelho, Cristina; Saraiva, Joana; Osório, Hugo; Antunes, Marília; Vaz, Fátima; Neves, Sofia; Pinto, Paula; Bárbara, Cristina; Penque, DeborahObstructive Sleep Apnea (OSA) is characterized by recurrent-episodes of apneas/hypopneas during sleep, leading to recurrent intermittent-hypoxia and sleep fragmentation. Non-treated OSA can result in cardiometabolic diseases. In this study, we applied a shotgun-proteomics strategy to deeper investigate the red blood cell-(RBC) homeostasis regulation in the context of OSA-severity and their response to six months of positive airway pressure (PAP)-treatment. RBC-samples from patients with Mild/Severe-OSA before/after-PAP treatment and patients as simple-snoring controls were selected. The mass-spectrometry raw-data was analysed by MaxQuant for protein identification/quantification followed by statistical Linear Models-(LM) and Linear Mixed Models-(LMM) to investigate OSA-severity effect and interaction with PAP, respectively. The functional/biological network analysis were performed by DAVID-platform. The results indicated that key-enzymes of the Embden-Meyerhof-Parnas (EMP)-glycolysis and pentose phosphate pathway-(PPP) were differentially changed in Severe-OSA, suggesting that the O2-dependent metabolic flux through EMP and PPP maybe compromised in these cells due to severe intermittent hypoxia/reoxygenation-induced oxidative-stress events in these patients. The Rapoport-Luebering-glycolytic shunt showed a significant downregulation across OSA-severity maybe to increase hemoglobin-O2 affinity to adapt to O2 low availability in the lung, although EMP-glycolysis showed decreased only in Severe-OSA. Proteins of the immunoproteasome were upregulated in Severe-OSA maybe to respond to severe oxidative-stress. In Mild-OSA, proteins related to the ubiquitination/neddylation-(Ub/Ned)-dependent proteasome system were upregulated. After PAP, proteins of Glycolysis and Ub/Ned-dependent proteasome system showed reactivated in Severe-OSA. In Mild-OSA, PAP induced upregulation of immunoproteasome proteins, suggesting that this treatment may increase oxidative-stress in these patients. Once validated these proteins maybe candidate biomarkers for OSA or OSA-therapy response.
- Assessing the role of children in the COVID-19 pandemic in Belgium using perturbation analysisPublication . Angeli, Leonardo; Caetano, Constantino Pereira; Franco, Nicolas; Coletti, Pietro; Faes, Christel; Molenberghs, Geert; Beutels, Philippe; Abrams, Steven; Willem, Lander; Hens, NielUnderstanding the evolving role of different age groups in virus transmission is essential for effective pandemic management. We investigated SARS-CoV-2 transmission in Belgium from November 2020 to February 2022, focusing on age-specific patterns. Using a next generation matrix approach integrating social contact data and simulating population susceptibility evolution, we performed a longitudinal perturbation analysis of the effective reproduction number to unravel age-specific transmission dynamics. From November to December 2020, adults in the [18, 60) age group were the main transmission drivers, while children contributed marginally. This pattern shifted between January and March 2021, when in-person education resumed, and the Alpha variant emerged: children aged under 12 years old were crucial in transmission. Stringent social distancing measures in March 2021 helped diminish the noticeable contribution of the [18, 30) age group. By June 2021, as the Delta variant became the predominant strain, adults aged [18, 40) years emerged as main contributors to transmission, with a resurgence in children’s contribution during September-October 2021. This study highlights the effectiveness of our methodology in identifying age-specific transmission patterns.
- Detection of Rickettsia in ticks using loop-mediated isothermal amplification (LAMP)Publication . Lansdell, Samantha; Hassan, Marwa M.; La Ragione, Roberto; Betson, Martha; Nuncio, MS; Lopes de Carvalho, Isabel; Zé-Zé, Líbia; de Sousa, Rita; Cutler, Sally; Davis, Joshua S.Objectives: The objective of the study was to develop a Loop-mediated Isothermal Amplification (LAMP) assay for screening of Rickettsia species circulating in ticks using the citrate synthase gene (gltA). The LAMP assay employed portable visualisation methods, making the assay more field-suitable. Furthermore, prior methods have not used gltA as the target, despite proven success in Polymerase Chain Reaction (PCR) methods. Methods: Using an alignment of 72 DNA sequences (comprised of 21 Rickettsia species) from GenBank we designed a novel set of gltA LAMP primers. Evaluation used DNA from 12 Rickettsia species as positive controls (extracted from cultures or naturally infected ticks) alongside a panel of negative controls representing different bacterial species. Subsequently this assay was used to screen 295 Ixodes ricinus and 24 I. hexagonus ticks collected from the UK (including northern and southern England and northern Scotland). Results: LAMP successfully detected 11 out of 12 (91.7%) Rickettsia species, excluding Rickettsia akari. Among 319 ticks collected in the UK, three were positive for Rickettsia (0.9%). All three positives were I. ricinus ticks, while none of the 24 I. hexagonus ticks were positive. Results were confirmed using a published PCR method. Sanger sequencing of PCR amplicons generated for each positive tick showed that they were all R. helvetica. Conclusions: This study introduces a novel field-applicable LAMP protocol for efficient Rickettsia screening in ticks to better assess its prevalence and consequent health risks. Furthermore, this assay has proven suitability for rickettsial detection in I. ricinus ticks, which has been reported as unsuccessful in previous European studies.
- Genetic variants in the IFNGR2 locus associated with severe chronic Q feverPublication . David, Susana; Castro, Liliana; Duarte, Elsa; Gaspar, Ulisses; Rodrigues, Maria Rosário da Costa; Cueto-Rojo, Maria Vanessa; Mendonça, Joana; Ferrão, José; Machado, Miguel; Poças, José; Lavinha, João; Vieira, Luís; Santos, Ana Sofia; ElsevierQ fever is a highly contagious zoonosis capable of causing large outbreaks of important health and economic consequences. Host genetic factors are believed to influence the development of severe chronic Q fever following the infection by the etiological agent, Coxiella burnetii. Targetted next generation sequencing (NGS) was performed in a case-control genetic association study on 53 confirmed Q fever cases, including 38 compatible with acute and 15 with chronic disease, and 29 samples from the general Portuguese population. Four SNPs in the IFNGR2 locus, rs78407108 G > A, rs17879956 C > T, rs7277167 C > T, and rs9974603 C > A, showed a statistically significant association to chronic Q fever, resisting the Bonferroni correction. These belonged to haplotypes significantly associated with chronic Q fever. The individual SNPs are referenced in the GTEx database as possible eQTLs. Given the direct bearing of IFNGR2 on IFN-γ signaling, the possible involvement of the associated variants with higher IFNGR2 expression could be in line with observations suggesting that IFN-γ production in chronic Q fever patients is significantly higher than in healthy controls. Further investigations are required to clarify the role of IFNGR2 signaling in association with chronic Q fever.
- Effectiveness of the XBB.1.5 COVID-19 Vaccines Against SARS-CoV-2 Hospitalisation Among Adults Aged ≥ 65 Years During the BA.2.86/JN.1 Predominant Period, VEBIS Hospital Study, Europe, November 2023 to May 2024Publication . Antunes, Liliana; Rojas-Castro, Madelyn; Lozano, Marcos; Martínez-Baz, Iván; Leroux-Roels, Isabel; Borg, Maria-Louise; Oroszi, Beatrix; Fitzgerald, Margaret; Dürrwald, Ralf; Jancoriene, Ligita; Machado, Ausenda; Petrović, Goranka; Lazar, Mihaela; Součková, Lenka; Bacci, Sabrina; Howard, Jennifer; Verdasca, Nuno; Basile, Luca; Castilla, Jesús; Ternest, Silke; Džiugytė, Aušra; Túri, Gergő; Duffy, Roisin; Hackmann, Carolin; Kuliese, Monika; Gomez, Verónica; Makarić, Zvjezdana Lovrić; Marin, Alexandru; Husa, Petr; Nicolay, Nathalie; Rose, Angela M.C.; VEBIS SARI VE network teamWe estimated the effectiveness of the adapted monovalent XBB.1.5 COVID-19 vaccines against PCR-confirmed SARS-CoV-2 hospitalisation during the BA.2.86/JN.1 lineage-predominant period using a multicentre test-negative case-control study in Europe. We included older adults (≥ 65 years) hospitalised with severe acute respiratory infection from November 2023 to May 2024. Vaccine effectiveness was 46% at 14-59 days and 34% at 60-119 days, with no effect thereafter. The XBB.1.5 COVID-19 vaccines conferred protection against BA.2.86 lineage hospitalisation in the first 4 months post-vaccination.
- Effectiveness of the XBB.1.5 COVID-19 Vaccines Against SARS-CoV-2 Hospitalisation Among Adults Aged ≥ 65 Years During the BA.2.86/JN.1 Predominant Period, VEBIS Hospital Study, Europe, November 2023 to May 2024Publication . Antunes, Liliana; Rojas-Castro, Madelyn; Lozano, Marcos; Martínez-Baz, Iván; Leroux-Roels, Isabel; Borg, Maria-Louise; Oroszi, Beatrix; Fitzgerald, Margaret; Dürrwald, Ralf; Jancoriene, Ligita; Machado, Ausenda; Petrović, Goranka; Lazar, Mihaela; Součková, Lenka; Bacci, Sabrina; Howard, Jennifer; Verdasca, Nuno; Basile, Luca; Castilla, Jesús; Ternest, Silke; Džiugytė, Aušra; Túri, Gergő; Duffy, Roisin; Hackmann, Carolin; Kuliese, Monika; Gomez, Verónica; Makarić, Zvjezdana Lovrić; Marin, Alexandru; Husa, Petr; Nicolay, Nathalie; Rose, Angela M. C.; VEBIS SARI VE network teamWe estimated the effectiveness of the adapted monovalent XBB.1.5 COVID-19 vaccines against PCR-confirmed SARS-CoV-2 hospitalisation during the BA.2.86/JN.1 lineage-predominant period using a multicentre test-negative case-control study in Europe. We included older adults (≥ 65 years) hospitalised with severe acute respiratory infection from November 2023 to May 2024. Vaccine effectiveness was 46% at 14-59 days and 34% at 60-119 days, with no effect thereafter. The XBB.1.5 COVID-19 vaccines conferred protection against BA.2.86 lineage hospitalisation in the first 4 months post-vaccination.
- Safety evaluation of pea fibre concentrate (FIPEA) as food additivePublication . EFSA Panel on Food Additives and Flavourings (FAF); Castle, Laurence; Andreassen, Monica; Aquilina, Gabriele; Bastos, Maria Lourdes; Boon, Polly; Fallico, Biagio; FitzGerald, Reginald; Frutos-Fernandez, Maria Jose; Grasl-Kraupp, Bettina; Gundert-Remy, Ursula; Gürtler, Rainer; Houdeau, Eric; Kurek, Marcin; Louro, Henriqueta; Morales, Patricia; Passamonti, Sabina; Barat Baviera, José Manuel; Degen, Gisela; Gott, David; Leblanc, Jean-Charles; Moldeus, Peter; Waalkens-Berendsen, Ine; Wölfle, Detlef; Gagliardi, Gabriele; Mech, Agnieszka; Smeraldi, Camilla; Tard, Alexandra; Zakidou, Panagiota; Ruggeri, LauraThe EFSA Panel on Food Additive and Flavourings (FAF Panel) provides a scientific opinion on the safety assessment of the proposed use of pea fibre concentrate (FIPEA) as a food additive. FIPEA is a powder consisting mainly of dietary fibres (i.e. pectin and hemicellulose), and low amounts of protein, derived from yellow pea (P. sativum). The manufacturing process includes extensive heat treatments, (e.g. > 100°C for more than 40 min), conditions which lead to inactivation of lectins, that in FIPEA do not pose a safety concern. A specific α‐amylase is used in the manufacturing, and this should be included in the definition of the proposed specifications. The Panel considered that the additional contribution of FIPEA to the total fibre intake in adults and toddlers would be acceptable considering the levels that are considered adequate by the NDA Panel. The Panel recommended to lower the specification limits proposed for the toxic elements. The solubility test indicates that the material does not require specific assessment at the nanoscale. No toxicological data have been submitted on FIPEA. The Panel considered that, similarly to water‐soluble soybean polysaccharides, FIPEA is not absorbed intact but undergoes extensive fermentation by the intestinal microbiota in humans and is not of genotoxic concern. Dry peas (raw material) are a staple food, with a very long history of safe use in the EU. FIPEA is extracted with hot water from the insoluble fibrous material of dehulled yellow peas, therefore the structure of the fibres is not chemically modified, and no new by‐products or components of toxicological concern are expected from the manufacturing process. The Panel concluded that there was no need for a numerical acceptable daily intake (ADI) and that pea fibre concentrate (FIPEA) as a new food additive does not raise a safety concern at the proposed use and use levels.
- Re‐evaluation of pullulan (E 1204) as a food additive and new application for its extension of usePublication . EFSA Panel on Food Additives and Flavourings (FAF); Castle, Laurence; Andreassen, Monica; Frutos Fernandez, Maria Jose; Aquilina, Gabriele; Bastos, Maria Lourdes; Boon, Polly; Fallico, Biagio; Fitzgerald, Reginald; Grasl-Kraupp, Bettina; Gundert-Remy, Ursula; Gürtler, Rainer; Houdeau, Eric; Kurek, Marcin; Louro, Henriqueta; Morales, Patricia; Passamonti, Sabina; Barat Baviera, José Manuel; Degen, Gisela; Gott, David; Leblanc, Jean-Charles; Moldeus, Peter; Waalkens-Berendsen, Ine; Wölfle, Detlef; Aguilera Entrena, Jaime; Gagliardi, Gabriele; Mech, Agnieszka; Medrano-Padial, Concepción; Lunardi, Simone; Rincon, Ana Maria; Smeraldi, Camilla; Tard, Alexandra; Ruggeri, LauraThe present opinion deals with the re‐evaluation of pullulan (E 1204) when used as a food additive and with the new application on the extension of use to several food categories. Pullulan (E 1204) is obtained by fermentation of a food‐grade hydrolysed starch with non‐genetically modified Aureobasidium pullulans ■■■■■. Based on the available information, the Panel considered that the manufacturing process of pullulan (E 1204) using this microorganism does not raise a safety concern. The Panel confirmed that pullulan (E 1204) is of no concern for genotoxicity. In vitro, pullulan (E 1204) is broken down by salivary and pancreatic amylase and intestinal iso‐amylase and it is further metabolised to short chain fatty acids in the colon by fermentation. Human adult volunteer studies suggested that effects of pullulan (E 1204) are similar to the effects of other poorly digestible carbohydrate polymers including modified celluloses and that mild undesirable gastrointestinal symptoms (i.e. abdominal fullness, flatulence, bloating and cramping) may occur at doses of 10 g pullulan per day and greater. The Panel compared the dose of 10 g pullulan per day with the dietary exposure estimates to pullulan (E 1204) in its currently permitted uses and considering the proposed changes to the currently permitted uses. The Panel concluded that there is no need for a numerical ADI for pullulan (E 1204) and there is no safety concern for the currently reported uses and use levels. Additionally, the Panel concluded that the exposure estimates considering the proposed changes to the currently permitted uses and use levels of pullulan (E 1204) are of no safety concern. The estimates for dietary exposure to pullulan (E 1204) indicate that individuals with a high level of exposure, principally coming from food supplements, may experience mild gastrointestinal symptoms at the currently reported uses and use levels.
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