National Institute of Health Dr. Ricardo Jorge
Repositório Científico do Instituto Nacional de SaúdeNot a member yet
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Adult cascade screening versus child reverse cascade screening in Familial Hypercholesterolemia
No full textFamilial hypercholesterolemia (FH) is a common inherited lipid disorder that predisposes to cardiovascular disease (CVD). Despite most cascade screening programs are initiated by adult index cases, reverse cascade screening pediatric index cases is starting to be described. Therefore, we aimed to assess the outcome of adult cascade screening and child reverse cascade screening strategies in families from the Portuguese FH Study (PFHS). The PFHS database was consulted, and 423 index cases genetically identified with FH (224 adults and 199 children) and their 997 relatives referred to the PFHS were analysed. From 224 adults with FH, 485 relatives were enrolled for cascade screening and 290 were identified with FH. From 199 paediatric cases with FH, 512 relatives were screened and 286 were identified with FH. Child reverse cascade screening presented a slightly higher diagnostic rate than adult cascade screening, 1.44 vs 1.29 new cases with FH per index case, and the age of the relatives identified was younger, 29 vs 37 years. For 94% of index children, relatives were referred (2.56 relatives per index), in contrast with the adult cohort whereas only 70% were referred with family-members (2.17 relatives per index). Overall, both screening approaches constitute valuable tools to identify new cases with FH, but the child reverse cascade screening creates the opportunity for more relatives to be tested at a younger age. It is crucial to improve relatives' recruitment rate since early identification allows a correct FH diagnosis and treatment to prevent CVD.N/
Functional characterization of two APOB variants from exon 29 found in individuals with clinical diagnosis of Familial Hypercholesterolemia
Get PDFFamilial hypercholesterolemia (FH) is an inherited lipid disorder characterized by increased levels of LDL cholesterol. About 5-10% of FH cases occur due to variants in the APOB gene, but these alterations can be a more common cause of FH than expected since most of APOB variants identified is still unknown their effect on the metabolism. The majority of the variants are missense but there are a few nonsense variants and small indels in exon 29 identified in individuals with hypercholesterolemia phenotype that can cause FH. The aim of this project was to functional characterize APOB variants from exon 29 identified in individuals referred to the Portuguese FH Study to assess if these are the genetic cause of disease.
LDL from index cases and relatives was isolated through sequential ultracentrifugation. ED-LDLR was purified from HEK293 cells transfected with the pcDNA3.1-EC-LDLR-His plasmid by affinity chromatography. Purified ED-LDLR fragments were coated onto 96-well plates and incubated with the different APOB variants. Antibodies were used for ligand detection, and absorbance was determined at 405 nm. CHO-ldlA7 cells were transfected with wt LDLR plasmid and incubated with FITC-labeled LDL to determine LDL binding and uptake by flow cytometry.
p.(Gln4316*) and p.(Glu4387Asnfs*7) alterations from exon 29 showed reduced affinity for the LDL receptor. Uptake and binding assays results were similar, so these variants may affect the binding of apoB to the LDL receptor. The alterations studied were not present in a normolipidemic panel.
APOB variants studied in this work produce truncated forms of apoB, but they are unlikely to lead to nonsense-mediated decay processes due to their location near the end of the gene. Functional studies can provide important evidence for variant pathogenicity assessment being these essential to provide an accurate diagnosis. These assays can confirm the clinical diagnosis by highlighting the cause of disease, and contribute to a personalized treatment and stratify patient associated cardiovascular risk.N/
Distress among healthcare professionals during the first two years of COVID-19 pandemic in Portugal
Get PDFThe COVID-19 pandemic increased existing psychosocial risk factors among healthcare professionals (HCPs). Integrated into a wider project, the main objective of this study was to characterize Portuguese HCPs mental health (MH), estimate the percentage of symptoms of anxiety, depression, post-traumatic stress disorder (PTSD), and burnout, and identify risk and protective factors. Methods: A cross-sectional online survey and a longitudinal assessment were conducted in 2020 (T0) and 2021 (T1). Sociodemographic and occupational variables, as well as protection behaviors and pandemic-context data were collected from a large non-probabilistic sample of HCPs in Portugal. MH outcomes were assessed using instruments with sound psychometric properties. Risk and protective factors were evaluated through simple and multiple logistic regression models. Results: A total of 2027 HCPs participated in the survey in T0; 1843 in T1. Despite the percentage of moderate to severe symptoms had decreased from T0 to T1: 26.1% and 23.3% for anxiety (T0 and T1, respectively; p = 0.028), 25.3% and 23.7% for depression, 22.7% and 19.1% for PTSD (p = 0.003), and 29.8% to 29.5% for burnout, a considerable proportion of HCPs reported symptoms of distress in both years. Being a woman, working in a COVID-19-treatment frontline position and the perception of work-life imbalance increased the odds of distress (in both T0 and T1), whilst high resilience, good social/family support, and hobbies/lifestyle maintenance were found to be MH protective factors. Conclusions: The longitudinal approach of our study allowed following-up changes in HCPs mental health and show that performing as an HCP during the pandemic may result in long-term effects on MH. Our results also provide evidence to support interventions targeting gender and professional sub-groups. Further studies are needed to understand the potential long-lasting psychological burden related to COVID-19 among HCPs.N/
First comparative genomic characterization of the MSSA ST398 lineage detected in aquaculture and other reservoirs
Get PDFStaphylococcus aureus ST398 can cause diseases in several different animals. In this study we analyzed ten S. aureus ST398 previously collected in three different reservoirs in Portugal (humans, gilthead seabream from aquaculture and dolphin from a zoo). Strains tested against sixteen antibiotics, by disk diffusion or minimum inhibitory concentration, showed decreased susceptibility to benzylpenicillin (all strains from gilthead seabream and dolphin) and to erythromycin with an iMLSB phenotype (nine strains), and susceptibility to cefoxitin (methicillin-susceptible S. aureus, MSSA). All strains from aquaculture belonged to the same spa type, t2383, whereas strains from the dolphin and humans belonged to spa type t571. A more detailed analysis using single nucleotide polymorphisms (SNPs)-based tree and a heat map, showed that all strains from aquaculture origin were highly related with each other and the strains from dolphin and humans were more distinct, although they were very similar in ARG, VF and MGE content. Mutations F3I and A100V in glpT gene and D278E and E291D in murA gene were identified in nine fosfomycin susceptible strains. The blaZ gene was also detected in six of the seven animal strains. The study of the genetic environment of erm(T)-type (found in nine S. aureus strains) allowed the identification of MGE (rep13-type plasmids and IS431R-type), presumably involved in the mobilization of this gene. All strains showed genes encoding efflux pumps from major facilitator superfamily (e.g., arlR, lmrS-type and norA/B-type), ATP-binding cassettes (ABC; mgrA) and multidrug and toxic compound extrusion (MATE; mepA/R-type) families, all associated to decreased susceptibility to antibiotics/disinfectants. Moreover, genes related with tolerance to heavy metals (cadD), and several VF (e.g., scn, aur, hlgA/B/C and hlb) were also identified. Insertion sequences, prophages, and plasmids made up the mobilome, some of them associated with ARG, VF and genes related with tolerance to heavy metals. This study highlights that S. aureus ST398 can be a reservoir of several ARG, heavy metals resistance genes and VF, which are essential in the adaption and survival of the bacterium in the different environments and an active agent in its dissemination. It makes an important contribution to understanding the extent of the spread of antimicrobial resistance, as well as the virulome, mobilome and resistome of this dangerous lineage.VS has her Ph.D. fellowship granted by FCT (Fundação para a
Ciência e a Tecnologia) with the reference SFRH/BD/133100/2017
co-financed by European Social Fund and the Operational Program for Human Capital (POCH), Portugal. This work was financial
supported with funding from FCT/MCTES (UIDB/00211/2020)
through national funds.info:eu-repo/semantics/publishedVersio
Neuron and Glial Cells Exposed to Cerium Dioxide Nanoparticles: Results from MTT and γH2AX Assays
Get PDFThis article belongs to the Proceedings of The 4th International Online Conference on Nanomaterials publicado em: Mater Proc. 2023;14(1):4. https://www.mdpi.com/2673-4605/14/1/4Proceeding PaperCerium dioxide nanoparticles (CeO2NP) show antioxidant enzyme-like properties and reactive oxygen species (ROS) scavenging activity, making them a promising material for potential therapeutic applications in neurodegenerative diseases. The objective of this work was to assess the biological behavior of CeO2NP in human SH-SY5Y neuronal and A172 glial cells by means of the MTT assay and the γH2AX assay. Despite the significant dose- and time-dependent NP internalization by both cell lines, nanoceria generally presented scarce cytotoxicity or genotoxicity, essentially restricted to the highest NP doses and longest exposure times. In conclusion, a high biocompatibility of CeO2NP was observed under the conditions tested.This research was funded by the Spanish Ministry of Science and Innovation MCIN/AEI/10.13039/501100011033, grant PID2020-114908GA-I00; Xunta de Galicia (ED431B 2022/16); CICA-Disrupting Project (2021SEM-B2); the Spanish Ministry of Education, Culture and Sport (BEAGAL18/00142 to V.V.); and the Portuguese Fundação para a Ciência e a Tecnologia (FCT) (SFRH/BPD/122112/2016 to A.T.R).info:eu-repo/semantics/publishedVersio
Mark-Release-Recapture of Aedes albopictus in Portugal: the influence of climatic factors
Get PDFAedes albopictus mosquitoes spread diseases like dengue, Zika, yellow fever, and chikungunya. The Sterile Insect Technique (SIT), which is based on production of male sterile mosquitoes through radiation and in the release in target areas, can be used as a prevention and control tool against Ae. albopictus native populations. Mark-Release-Recapture trials (MRR) estimate the number of mosquitoes to release during the SIT, but weather conditions can affect results. Thus, we aimed to understand the influence of climatic factors on mosquito collection. MRRs were conducted in October 2022 in Faro, Southern Portugal. Mosquito sterilisation, marking and transport followed protocols from the International Atomic Energy Agency. Releases occurred weekly for three consecutive weeks at two different sites, and mosquitoes were collected using Human Landing Collections (HLC) one, two, four and six days after release. Climatic data, such as temperature, humidity, wind intensity (moderate 15-35 km/h vs weak <15 km/h) and precipitation, were obtained from the Portuguese Institute of the Sea and the Atmosphere. We used generalised linear models with a negative binomial family and log function to estimate factors associated with the number of captured mosquitoes, estimated prevalence ratios (PRs), and the 95% confidence intervals (CI). We released 84 000 marked sterile male mosquitoes and recaptured 528 marked males (0.7%) by HLC. The prevalence of captured mosquitoes was 23% lower when the wind intensity was moderate compared with a weak wind intensity (PR: 0.77, 95% CI: 0.61−0.98). We did not find a statistically significant association between the number of captured mosquitoes and humidity (PR: 0.98, 95% CI: 0.96−1.00), temperature (PR: 1.03, 95% CI: 0.83−1.28) and precipitation (PR: 1.07, 95% CI: 0.60−2.03). Conclusion: It is crucial to plan MRRs carefully and consider weather conditions during their execution to avoid underestimating the population needed for the SIT and compromising the effectiveness of this control method.This work has been supported by IAEA project RER5026 Enhancing the Capacity to Integrate Sterile Insect Technique in the Effective Management of Invasive Aedes Mosquitoes. Patrícia Soares received funding from CEECINST/00049/2021/CP2817/CT0001.N/
Contaminação de superfícies por fungos em habitação: Um risco para a saúde
Get PDFIntrodução: Existe suficiente evidência epidemiológica de que os ocupantes de edifícios com problemas de humidade excessiva e contaminação de superfícies por fungos, têm um risco acrescido de desenvolver sintomas respiratórios, infeções respiratórias e de exacerbar crises de asma. No interior de edifícios, a relativa abundância de diferentes espécies de fungos tende a seguir o padrão exterior, embora as concentrações interiores sejam geralmente mais baixas. Segundo o Organização Mundial de Saúde a existência de crescimento visível de bolores em superfícies é uma forte evidência da exposição humana a estes agentes (1).
Objetivo: O objetivo deste estudo é evidenciar que o aparecimento de fungos em superfícies está relacionado com o aumento da concentração de esporos suspensos no ar e/ou de colónias fúngicas nas superfícies e que as espécies isoladas são, na sua maioria, semelhantes.
Metodologia: Foram colhidas amostras de ar no exterior (referência) e no interior de uma habitação T2 situada no piso térreo onde, na sequência de uma rutura de canalização de águas residuais, se observou o desenvolvimento visível de bolor nas paredes, em novembro de 2022. A amostragem em duplicado de fungos viáveis no ar foi realizada utilizando o amostrador MAS-100 e placas de Agar Extrato de Malte suplementadas com cloranfenicol. Nas superfícies foram realizadas amostragens de fungos utilizando dois métodos, placas de contacto e zaragatoas posteriormente inoculadas em Agar de Extrato de Malte suplementadas com cloranfenicol. Resultados: As concentrações de fungos obtidas no ar interior da habitação, quarto e escritório, foram superiores a 10512 UFC/m3 ultrapassando as concentrações do exterior de 1356 UFC/m3. Das colónias isoladas no interior 46% têm potencial patogénico. Foram encontradas 17 estirpes diferentes (7 espécies e 10 géneros) no interior, incluindo no ar e nas superfícies e 13 (3 espécies e 10 géneros) no exterior.
Conclusões: Os resultados revelaram que as concentrações de fungos obtidas no interior da habitação são no mínimo 8 vezes superiores às do Exterior, tomadas como referência. A presença de espécies como Aspergillus glaucus, Exophiala dermatitidis, Conibiobolus coronatus apenas presentes nas amostras do interior, apresentam potencial patogénico, podendo causar osteomielite e artrite séptica, infeções cutâneas ou pulmonares, doença rinofacial crónica (2, 3). Somos levados a concluir que os esporos fúngicos encontrados no ar interior têm origem nas superfícies contaminadas em virtude do excesso de água.
O ar interior é um importante determinante de saúde para os ocupantes pelo que é urgente identificar, mitigar e prevenir situações de humidade persistente que potenciam o crescimento microbiano em superfícies.N/
Ethics and legal requirements for data linkage in 14 European countries for children with congenital anomalies
Get PDFIntroduction: Linking healthcare data sets can create valuable resources for research, particularly when investigating rare exposures or outcomes. However, across Europe, the permissions processes required to access data can be complex. This paper documents the processes required by the EUROlinkCAT study investigators to research the health and survival of children with congenital anomalies in Europe.
Methods: Eighteen congenital anomaly registries in 14 countries provided information on all the permissions required to perform surveillance of congenital anomalies and to link their data on live births with available vital statistics and healthcare databases for research. Small number restrictions imposed by data providers were also documented.
Results: The permissions requirements varied substantially, with certain registries able to conduct congenital anomaly surveillance as part of national or regional healthcare provision, while others were required to obtain ethics approvals or informed consent. Data linkage and analysis for research purposes added additional layers of complexity for registries, with some required to obtain several permissions, including ethics approvals to link the data. Restrictions relating to small numbers often resulted in a registry's data on specific congenital anomalies being unusable.
Conclusion: The permissions required to obtain and link data on children with congenital anomalies varied greatly across Europe. The variation and complexity present a significant obstacle to the use of such data, especially in large data linkage projects. Furthermore, small number restrictions severely limited the research that could be performed for children with specific rare congenital anomalies.info:eu-repo/semantics/publishedVersio
Diversity of Human T-Lymphotropic Virus Type 1 Cosmopolitan Subtype (HTLV-1a) Circulating in Infected Residents in Portugal
No full textHuman T-cell lymphotropic virus type 1 (HTLV-1) prevalence in Portugal is low and mainly affects immigrants from endemic areas where human immunodeficiency virus (HIV) infection represents a public health problem. Despite the majority of HTLV-1-infected individuals remains asymptomatic, severe pathologies may develop after prolonged viral persistence, namely an aggressive form of leukemia. An increased mortality rate and faster progression to death is often related to HTLV-1/HIV coinfection. Nevertheless, studies showed that some antiretrovirals used in HIV treatment lead to a positive immune response against HTLV-1. This study aimed to analyze epidemiological and clinical data, and to assess the diversity of HTLV-1 strains circulating in infected residents diagnosed in the Portuguese national reference laboratory between 2010 and 2021. Long terminal repeat and env proviral sequences derived from 20 individuals were used to generate phylogenetic trees along with multiples reference sequences from different geographic origins retrieved from the database. Three samples belong to Portuguese natives and 17 belong to immigrants: 15 from several countries of Africa, 1 from South America, and 1 from Europe; 6 patients (30%, mean age 40.3 years) showed HTLV-1-related diseases, and 6 (30%, mean age 45.2 years) were coinfected with HIV/AIDS. The results show that the Cosmopolitan subtype is circulating in Portugal, with 10 sequences being classified as subgroup A, that include Portuguese and natives from S. Tomé and Príncipe with a mean age of 39.4 years, and 10 sequences that segregated into the Senegal cluster derived from natives born in Guinea-Bissau with a mean age of 43.5 years. A high proportion of HTLV-1-related diseases and HIV/AIDS coinfection was observed. Risk behavior practices and the absence of specific control measures, including diagnostic and treatment, may contribute to a silent dissemination of a broad diversity of HTLV-1 strains and, therefore, the increased rate of progression to debilitating diseases. In this manner, an early diagnostic and a molecular surveillance of HTLV-1 transmission remains necessary in Portugal.info:eu-repo/semantics/publishedVersio
Exploring an antisense oligonucleotide exon-skipping therapeutic strategy for Mucolipidosis II
No full textIntroduction: Mucolipidosis II (ML II) is a Lysosomal Storage Disorder caused by the
deficiency of the enzyme GlcNAc-1-phosphotransferase, which is responsible for the
Mannose-6-Phosphate marker addition to lysosomal enzymes. Of all ML II mutations,
the c.3503_3504delTC in GNPTAB exon 19 is the most frequent, making it a good
target for a personalized therapy. Here, we explored an innovative therapeutic
strategy based on the use of antisense oligonucleotides (ASOs) for ML II. Previously, on
ML II patients’ fibroblasts, ASOs were used to skip exon 19 of the GNPTAB pre-mRNA,
successfully resulting in the production of an in-frame mRNA. Now, our aim is to
analyze if these results are translated to the enzymatic and cellular phenotype level.N/