National Institute of Health Dr. Ricardo Jorge
Repositório Científico do Instituto Nacional de SaúdeNot a member yet
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How DIS3L2 meets NMD-targets: I’m really into “U”!
No full textThe nonsense-mediated mRNA decay (NMD) pathway selectively degrades mRNAs carrying a premature translation-termination codon but also regulates the abundance of a large number of physiological RNAs that encode full-length proteins. Also, NMD regulates the levels of many physiological PTC-free mRNAs that encode full-length proteins. In human cells, NMD-targeted mRNAs are degraded by endonucleolytic cleavage and exonucleolytic degradation from both 5’ and 3’ ends. This is achieved by a process not yet completely understood that promotes the decay of the mRNAs in 5’-to-3’ and 3’-to-5’ by the XRN1 and exosome, respectively. In yeast, Dis3/Rrp44 protein is the catalytic subunit of the exosome, but in humans, there are three known paralogues of this enzyme: DIS3, DIS3L1, and the Perlman syndrome-associated exoribonuclease DIS3L2. Conversely, to its counterparts, DIS3L2 activity is independent of the exosome. In order to unveil the role of DIS3L2 in NMD, we performed its knockdown in HeLa cells and measured the mRNA levels of various natural NMD-targets. Our results show that DIS3L2 is involved in NMD-targets decay. Besides that, DIS3L2 acts directly on NMD-targets and interacts with the key NMD factor UPF1. We also show that DIS3L2-mediated decay depends on the activity of the terminal uridylyl transferases (TUTases) 4 and 7, which adds non-templated uridines to the mRNAs 3’ end, marking these mRNAs for DIS3L2 degradation. Together, our findings establish a direct role of DIS3L2 in NMD in an uridylation-dependent manner.This work was partially supported by Fundacão para a Ciência e a Tecnologia (PTDC/BIM-MEC/3749/2014 and UID/MULTI/04046/2013 to BioISI from FCT/MCTES/PIDDAC).N/
Gene expression regulation by upstream open reading frames in colorectal cancer
No full textColorectal cancer (CRC) has a high incidence and mortality rates worldwide. Its carcinogenesis process is characterized by a continuous accumulation of genetic alterations that changes the overall gene expression profiles. Those alterations have been studied by microarray and RNA sequencing that measure the abundance of mRNA but do not provide information on protein synthesis, a step closer to the end-point of gene expression. Ribosome profiling (RiboSeq) emerges to monitor in vivo translation by deep sequencing of ribosome-protected mRNA fragments (RPFs). This technique detects ribosomes outside of known protein-coding regions, identifying translation of upstream open reading frames (uORFs) within 5’ untranslated regions (5’UTRs). The aim of this work is to determine the role of specific uORFs in CRC tumorigenesis. For that, we looked for potential uORFs-containing targets based in the 5’UTR RPFs occupancy from RiboSeq data from different cancer cell lines already available. We chose ABCE1, PAIP2, eIF4G2 and eIF2A as uORFs-containing mRNAs. Gene ontology analyses revealed an important role in translational control for the proteins encoded by these transcripts. By semi-quantitative RT-PCR, ABCE1 transcript is shown down-regulated in HCT116 cells in comparison to the non-neoplasic colorectal cell line (NCM460). To analyze the role of such uORFs in translational regulation and their biological function at the level of cell viability and proliferation, and acquisition of CRC features, a reporter plasmid was constructed carrying the ABCE1 5’UTR fused to the Firefly luciferase (Fluc) ORF (pGL2-ABCE1). Each one of the five upstream AUGs in ABCE15’UTR was mutated to obtained constructs with non-functinal uORFs and only one functional uORF. HCT116 cells were transiently transfected with pGL2-ABCE1 or each one of the above mentioned constructs. Fluc expression and activity was assessed by Western blot and luminometry assays, respectively. Results show a decrease in the translational efficiency of Fluc by pGL2-ABCE1. Moreover, the construct carrying only the uORF3 functional exhibits a stronger repression efficacy compared to pGL2-ABCE1 and the other constructs.Work partially supported by UID/MULTI/04046/2013 centre grant from FCT, Portugal (to BioISI). J.F.P.S., is recipient of a fellowship from BioSys PhD programme (Ref SFRH/BD/106081/2015) from FCT (Portugal).N/
The Functional Landscape Of Coding Variation In The Familial Hypercholesterolemia Gene LDLR
No full textVariants in the familial hypercholesterolemia gene -the most important genetic driver of cardiovascular disease-can raise circulating low-density lipoprotein (LDL) cholesterol concentrations and increase the risk of premature atherosclerosis. Definitive classifications are lacking for nearly half of clinically encountered missense variants, limiting interventions that reduce disease burden. Here, we tested the impact of ~17,000 (nearly all possible) missense coding variants on both LDLR cell-surface abundance and LDL uptake, yielding sequence-function maps that recapitulate known biochemistry, offer functional insights, and provide evidence for interpreting clinical variants. Functional scores correlated with hyperlipidemia phenotypes in prospective human cohorts and augmented polygenic scores to improve risk inference, highlighting the potential of this resource to accelerate familial hypercholesterolemia diagnosis and improve patient outcomes.Funding: R01HL164675 (to F.P.R., D.M.R., E.A.A., C.A.M., V.N.P., A.M.G., and B.M.K.) The One Brave Idea Initiative (jointly funded by the American Heart Association, Verily Life Sciences LLC, and Astra-Zeneca, Inc. with additional support from Quest Diagnostics) (to C.A.M. and F.P.R.). Canada Foundation for Innovation (to F.P.R.). Canadian Institutes of Health Research Foundation Grant FDN159926 (to F.P.R.). The Canada Excellence Research Chairs Program (to F.P.R.). University of Toronto Precision Medicine Initiative (PRiME) Fellowship PRMF2022 (to D.R.T.) National Human Genome Research Institute of the National Institutes of Health Center of Excellence in Genomic Science Initiative RM1HG010461 (to D.M.F. and F.P.R.). NHGRI Impact of Genomic Variation on Function Initiative UM1HG019189 (to F.P.R.). EU Horizon RIA: 101155885-2, FH-EARLY (to M.B. and S.G.P.). La Caixa Foundation (LCF/PR/HP23/52330032) (to M.B. and S.G.P.). NIH from R01DK106236, P30 DK116074 (to the Stanford Diabetes Research Center), R01DK116750, R01DK137889, R01DK120565 (to J.W.K), and NHGRI K08HG014001 (to M.C.L.) The All of Us Research Program is supported by the National Institutes of Health, Office of the Director: Regional Medical Centers: 1 OT2 OD026549; 1 OT2 OD026554; 1 OT2 OD026557; 1 OT2 OD026556; 1 OT2 OD026550; 1 OT2 OD026552; 1 OT2 OD026553; 1 OT2 OD026548; 1 OT2 OD026551; 1 OT2 OD026555; IAA#: AO 16037; Federally Qualified Health Centers: HHSN 26320160085U; Data and Research Center; 5 U2C OD023196; Biobank: 1 U24 OD023121; The Participant Center: U24 OD023176; Participant Technology Systems Center: 1 U24 OD023163; Communications and Engagement: 3 OT2 OD023205; 3 OT2 OD023206; and Community Partners: 1 OT2 OD025277; 3 OT2 OD025315; 1 OT2 OD025337; OT2 OD025276
Emerging Trends in Active Packaging for Food: A Six-Year Review
No full text(This article belongs to the Special Issue Active Packaging in Food Storage: From Development to Utilization—2nd Edition)The development of active food packaging has evolved rapidly in recent years, offering innovative solutions to enhance food preservation and safety while addressing sustainability challenges. This review compiles and analyzes recent advancements (2019–2024) in release-type active packaging, focusing on essential oils, natural extracts, and phenolic compounds as active agents. Primarily plant-derived, these compounds exhibit significant antioxidant and antimicrobial activities, extending shelf life and enhancing food quality. Technological strategies such as encapsulation and polymer blending have been increasingly adopted to overcome challenges related to volatility, solubility, and sensory impact. Integrating bio-based polymers, including chitosan, starch, and polylactic acid, further supports the development of environmentally friendly packaging systems. This review also highlights trends in compound-specific research, release mechanisms, and commercial applications, including a detailed analysis of patents and case studies across various food matrices. These developments have already been translated into practical applications, such as antimicrobial sachets for meat and essential oil-based pads for fresh produce. Moreover, by promoting the valorization of agro-industrial by-products and the use of biodegradable materials, emission-type active packaging contributes to the principles of the circular economy. This comprehensive overview underscores the potential of natural bioactive compounds in advancing sustainable and functional food packaging technologies.CardAPium project, funded by Portuguese national funds (FCT/MCTES, Fundação para a Ciência e Tecnologia and Ministério da Educação, Ciência e Inovação) through the grant 2023.15813.PEX (https://doi.org/10.54499/2023.15813.PEX); FCT—Fundação para a Ciência e Tecnologia/MECI through the MEtRICs research unit (UID/04077/2020): Mechanical Engineering and Resource Sustainability Center).
Ph.D. Grant 2021.08154.BD (https://doi.org/10.54499/2021.08154.BD)
Characteristics and incidence trends of adults hospitalized with community-acquired pneumonia in Portugal, pre-pandemic
No full textCommunity-acquired pneumonia (CAP) is a major cause of hospitalization that leads to substantial morbidity, mortality, and costs. Evaluating CAP trends over time is important to understand patterns and the impact of public health interventions. This study aims to describe the characteristics and trends in the incidence of adults hospitalized with CAP in Portugal between 2010 and 2018. In this study, we included hospitalization data, prevalence of comorbidities, and population data. CAP hospitalizations of adults (≥18y) living in mainland Portugal discharged from public hospitals were identified using ICD-9-CM or ICD-10-CM codes. Based on previous CAP studies, we selected nine relevant comorbidities. We described the frequency and incidence of CAP hospitalizations per sex, age group, comorbidity, and year of discharge. Trends were explored using Joinpoint regression. We observed 470,545 CAP hospitalizations falling into the 2010-18 period. The majority were males (54.8%) and aged ≥75 years (65.3%). Most often recorded comorbidities were congestive heart failure (26.4%), diabetes (25.5%), and chronic pulmonary disease (19.2%). The Joinpoint regression identified a gradual decline in the incidence rates of CAP hospitalizations for both sexes and all age groups. Of the nine comorbidities selected, seven showed a progressive increase in incidence rates followed by a subsequent decline (all except HIV/AIDS and chronic renal disease). Our findings offer valuable insights for selecting priority groups for public health interventions and design strategies to mitigate the burden of CAP.This study was conducted as a research collaboration between the NOVA National School of Public Health (sponsor) and Pfizer (funder)
Newborn Screening for Sickle Cell Disease: Results from a Pilot Study in the Portuguese Population
No full textThe Portuguese Newborn Screening Program currently includes 28 pathologies: congenital hypothyroidism, cystic fibrosis, 24 inborn errors of metabolism, sickle cell disease and spinal muscular atrophy. This pilot study for sickle cell disease newborn screening, including 188,217 samples, was performed between May 2021 and December 2023, with phase I, including 24,130 newborns, in the Lisbon and Setubal districts and phase II, including 164,087 newborns, in the whole country. DBS samples were analyzed through capillary electrophoresis. In phase I, a high birth incidence of sickle cell disease was found (1:928 NBs), resulting from the identification of 24 HbSS and 2 HbSC patients. This birth incidence decreased but remained significant when the pilot study for sickle cell disease newborn screening was expanded to a national level, with the identification of 67 sickle cell disease patients (59 HbSS and 8 HbSC), revealing a birth incidence of 1:2449 NBs. These data suggest that this condition is becoming increasingly relevant in Portugal, thus reflecting a general European trend, where sickle cell disease is already recognized as a public health problem. Therefore, it highlights the importance of its integration into the Portuguese National Newborn Screening Program panel in January 2024, thus allowing the early identification and clinical follow-up of these patients.The funding for the pilot study was The funding for the pilot study was provided by the NATIONAL INSTITUTE OF HEALTH DOCTOR RICARDO JORGE, which supports the National Neonatal Screening Program.provided by the NATIONAL INSTITUTE OF HEALTH DOCTOR RICARDO JORGE, which supports the National Neonatal Screening Program
Alternative protein sources: insect consumption and the promotion of sustainable food systems
No full textA crescente pressão exercida sobre os sistemas alimentares globais e os impactos ambientais da produção pecuária impulsionaram a procura por fontes de proteína alternativas sustentáveis. Os insetos comestíveis surgem como uma alternativa promissora, com elevado valor nutricional e reduzido impacto ambiental.
Este estudo analisa a literatura disponível sobre a utilização de insetos como fonte de proteína para consumo humano.
Espécies como Tenebrio molitor (tenébrio) e Acheta domesticus (grilo doméstico) apresentam elevada quantidade de proteína, lípidos, vitaminas, minerais e fibra, podendo substituir, parcialmente, as proteínas convencionais. A produção de insetos requer menos água e terra e gera menores emissões de gases de efeito estufa do que a produção pecuária convencional, e a sua capacidade de valorizar subprodutos agroalimentares contribui para a economia circular.
Contudo, a bioacumulação de contaminantes e a repulsa cultural por parte dos consumidores constituem barreiras à adoção generalizada do consumo de insetos, exigindo boas práticas de produção e estratégias para aumentar a aceitação por parte do consumidor.
Os insetos representam, assim, uma fonte proteica sustentável e eficiente, capaz de diversificar a alimentação e reduzir a pressão sobre os recursos naturais, consolidando o seu papel em sistemas alimentares resilientes e sustentáveis, alinhados com o conceito de Uma Só Saúde.The growing pressure on global food systems and the environmental impacts of livestock production have driven the search for alternative, sustainable protein sources. Edible insects have become a promising option, offering high nutritional value and a low environmental footprint.
This review examines the available literature on the use of insects as a protein source for human consumption.
Species such as Tenebrio molitor (mealworms) and Acheta domesticus (house crickets) are rich in protein, lipids, vitamins, minerals, and fibre, and can partially replace conventional proteins. Insect production requires less water and land and generates lower greenhouse gas emissions than traditional livestock farming, while their ability to utilise agri-food by-products contributes to the circular economy. However,
the bioaccumulation of contaminants and cultural resistance among consumers remain obstacles to widespread adoption, calling for good product ion practices and strategies to improve consumer acceptanceJoana Oliveira é financiada pela Fundação para a Ciência e a Tecnologia (FCT), Lisboa, Portugal, através de uma bolsa de doutoramento (referência 2022.13540.BDANA; https://doi.org/10.54499/2022.13540.BDANA). Este trabalho foi financiado pelos projetos: “InsectERA” (n.º C644917393-00000032), no âmbito dos WPs InBioremediation e One Health, com o apoio do Fundo Europeu Next Generation EU e do Plano de Recuperação e Resiliência (PRR) de Portugal, no quadro da linha de incentivo “Agendas para a Inovação Empresarial”, através do regime de financiamento C5 —Capitalização e Inovação Empresarial; CiiEM Investiga “FlyWaste” através do projeto 10.54499/UIDB/04585/2020, financiado pela FCT
