Browsing by Issue Date, starting with "2021-04"
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- Integration of point mutation and protein-protein interaction network analyses in Autism Spectrum DisorderPublication . Vilela, Joana; Martiniano, Hugo; Marques, Ana Rita; Santos, João; Rasga, Célia; Oliveira, Guiomar; Vicente, Astrid M.Objective: To identify neurotransmission and synaptic (NS) genes that play a role in ASD and address the impact of ultra-rare variants in these genes in ASD, linking these mutations to the biological pathways affected.
- Mutation-adapted U1snRNA as a therapeutic strategy for Mucopolysaccharidosis IIIC: in vitro and in vivo studiesPublication . Gonçalves, Mariana; Matos, Liliana; Santos, Juliana Inês; Coutinho, Maria Francisca; Prata, Maria João; Pires, Maria João; Oliveira, Paula; Alves, SandraA significant number of mutations that change the splicing process and lead to aberrant mRNA production have been identified in Lysosomal Storage Disorders (LSDs). Mucopolysaccharidosis type IIIC (MPS IIIC) is a very rare LSD caused by mutations in the HGSNAT gene which encodes an enzyme involved in heparan sulphate degradation. Splicing mutations are one of the most frequent (~20%) genetic defects in MPS IIIC. Around 55% correspond to 5' splice-site (ss) mutations thus constituting a good target for splicing therapeutics. Recently, we have demonstrated that a modified U1snRNA vector designed to improve the definition of the HGSNAT exon 2 5’ss can restore splicing impaired by the mutation c.234+1G>A. Currently, our goal is to evaluate in vivo the therapeutic potential of that modified U1 snRNA by testing it in mice expressing the human splicing defect. For this purpose, two full-length constructs were generated by cloning the wt or the mutated HGSNAT splicing-competent cassettes in the pcDNA 3.1 vector. Then, the wt and the mutant constructs were transfected in Hep3B and COS-7 cells. Both minigenes reproduce the healthy control and patient cDNA’s splicing pattern. Therefore, they were used to generate C57BL/6 mice expressing the wt (c.241+1G) or mutant (c.234+1A) alleles in the liver. These mice can be used for testing the modified U1 snRNA efficacy in vivo. Thus, wt or mutant minigenes were administrated in mice by hydrodynamic injection following a protocol described by Balestra et al. After 48h, animals were sacrificed, the liver was collected, and the molecular analysis was performed. Preliminary results showed the expression of the mutant construct in the liver of at least one animal. Thus, further tests will be carried out to optimize experimental conditions, by testing other forms of minigenes administration and using other mice strains.
- Characteristics of SARS-CoV-2 variants of concern B.1.1.7, B.1.351 or P.1: data from seven EU/EEA countries, weeks 38/2020 to 10/2021Publication . Funk, Tjede; Pharris, Anastasia; Spiteri, Gianfranco; Bundle, Nick; Melidou, Angeliki; Carr, Michael; Gonzalez, Gabriel; Garcia-Leon, Alejandro; Crispie, Fiona; O’Connor, Lois; Murphy, Niamh; Mossong, Joël; Vergison, Anne; Wienecke-Baldacchino, Anke K.; Abdelrahman, Tamir; Riccardo, Flavia; Stefanelli, Paola; Di Martino, Angela; Bella, Antonino; Lo Presti, Alessandra; Casaca, Pedro; Moreno, Joana; Borges, Vítor; Isidro, Joana; Ferreira, Rita; Gomes, João Paulo; Dotsenko, Liidia; Suija, Heleene; Epstein, Jevgenia; Sadikova, Olga; Sepp, Hanna; Ikonen, Niina; Savolainen-Kopra, Carita; Blomqvist, Soile; Möttönen, Teemu; Helve, Otto; Gomes-Dias, Joana; Adlhoch, CorneliaWe compared 19,207 cases of SARS-CoV-2 variant B.1.1.7/S gene target failure (SGTF), 436 B.1.351 and 352 P.1 to non-variant cases reported by seven European countries. COVID-19 cases with these variants had significantly higher adjusted odds ratios for hospitalisation (B.1.1.7/SGTF: 1.7, 95% confidence interval (CI): 1.0-2.9; B.1.351: 3.6, 95% CI: 2.1-6.2; P.1: 2.6, 95% CI: 1.4-4.8) and B.1.1.7/SGTF and P.1 cases also for intensive care admission (B.1.1.7/SGTF: 2.3, 95% CI: 1.4-3.5; P.1: 2.2, 95% CI: 1.7-2.8).
- Antisense oligonucleotide exon-skipping as a therapeutic approach for Mucolipidosis type II α/β: in vitro and in vivo studiesPublication . Gonçalves, Mariana; Matos, Liliana; Santos, Juliana Inês; Coutinho, Maria Francisca; Prata, Maria João; Pires, Maria João; Oliveira, Paula; Alves, SandraGenetic therapy directed towards the correction of RNA mis-splicing is being investigated at basic research and in late-stage clinical trials. Many mutations that change the normal splicing pattern and lead to aberrant mRNA production have been identified in Lysosomal Storage Disorders (LSDs). Mucopolysaccharidosis IIIC (MPS IIIC) is one of those LSDs caused by mutations in the HGSNAT gene that encodes an enzyme involved in heparan sulphate degradation. Splicing mutations are one of the most frequent (~20%) genetic defects in MPS IIIC. Approximately 55% correspond to 5' splice-site (ss) mutations thus constituting a good target for mutation-specific therapeutic approaches. Recently, we have demonstrated in fibroblast cells that a modified U1 snRNA vector designed to improve the definition of exon 2 5’ss of the HGSNAT can restore splicing impaired by the mutation c.234+1G>A. Currently, our goal is to evaluate in vivo the therapeutic potential of that modified U1 snRNA by testing it in mice expressing the human splicing defect. For this purpose, two full-length constructs were generated by cloning the wild-type (wt) or the mutated HGSNAT splicing-competent cassettes in the pcDNA 3.1 vector. Then, in an in vitro assay, the wt or mutated construct was transfected in Hep3B and COS-7 cells. After molecular analysis it was observed that both minigenes reproduce the healthy control and patient cDNA’s splicing pattern. Therefore, both constructs were used to generate mice of the C57BL/6 strain expressing the human mutation c.234+1G>A in the liver and test its modified U1-mediated rescue in vivo. Wt or mutant minigenes were administrated in mice by hydrodynamic injection following a reported protocol(1). After 48 hours animals were sacrificed, the liver was collected and molecular analysis was performed. Preliminary results showed expression of the HGSNAT cDNA from the mutant construct in the liver of at least one animal. Thus, further tests will be carried out to optimize some limiting points, such as the administration of the minigenes (e.g. increase of injection volume from 7% to 8-9% of mice body weight; inclusion of an in vivo transfection reagent to enhance delivery efficiency) and the use of other mice strain. 1. Balestra D, et al. (2014) J Thromb Haemost 12(2):177–185.
- miRNA target-binding sites as regulators of genes involved in the lipid metabolism might explain the hypercholesterolaemia in FH patientsPublication . Medeiros, A.M.; Enguita, F.J.; Bourbon, M.Aims to analyze miRNAs target sites as regulators of genes involved in the lipid metabolism in FH mutation-negative patients.
- Ambient carbon monoxide and daily mortality: a global time-series study in 337 citiesPublication . Chen, Kai; Breitner, Susanne; Wolf, Kathrin; Stafoggia, Massimo; Sera, Francesco; Vicedo-Cabrera, Ana M.; Guo, Yuming; Tong, Shilu; Lavigne, Eric; Matus, Patricia; Valdés, Nicolás; Kan, Haidong; Jaakkola, Jouni J.K.; Ryti, Niilo R.I.; Huber, Veronika; Scortichini, Matteo; Hashizume, Masahiro; Honda, Yasushi; Nunes, Baltazar; Madureira, Joana; Holobâcă, Iulian Horia; Fratianni, Simona; Kim, Ho; Lee, Whanhee; Tobias, Aurelio; Íñiguez, Carmen; Forsberg, Bertil; Åström, Christofer; Ragettli, Martina S-; Guo, Yue-Liang Leon; Chen, Bing-Yu; Li, Shanshan; Milojevic, Ai; Zanobetti, Antonella; Schwartz, Joel; Bell, Michelle L-; Gasparrini, Antonio; Schneider, AlexandraBackground: Epidemiological evidence on short-term association between ambient carbon monoxide (CO) and mortality is inconclusive and limited to single cities, regions, or countries. Generalisation of results from previous studies is hindered by potential publication bias and different modelling approaches. We therefore assessed the association between short-term exposure to ambient CO and daily mortality in a multicity, multicountry setting. Methods: We collected daily data on air pollution, meteorology, and total mortality from 337 cities in 18 countries or regions, covering various periods from 1979 to 2016. All included cities had at least 2 years of both CO and mortality data. We estimated city-specific associations using confounder-adjusted generalised additive models with a quasi-Poisson distribution, and then pooled the estimates, accounting for their statistical uncertainty, using a random-effects multilevel meta-analytical model. We also assessed the overall shape of the exposure-response curve and evaluated the possibility of a threshold below which health is not affected. Findings: Overall, a 1 mg/m3 increase in the average CO concentration of the previous day was associated with a 0·91% (95% CI 0·32-1·50) increase in daily total mortality. The pooled exposure-response curve showed a continuously elevated mortality risk with increasing CO concentrations, suggesting no threshold. The exposure-response curve was steeper at daily CO levels lower than 1 mg/m3, indicating greater risk of mortality per increment in CO exposure, and persisted at daily concentrations as low as 0·6 mg/m3 or less. The association remained similar after adjustment for ozone but was attenuated after adjustment for particulate matter or sulphur dioxide, or even reduced to null after adjustment for nitrogen dioxide. Interpretation: This international study is by far the largest epidemiological investigation on short-term CO-related mortality. We found significant associations between ambient CO and daily mortality, even at levels well below current air quality guidelines. Further studies are warranted to disentangle its independent effect from other traffic-related pollutants.
- Essential Oils from Plants: Industrial Applications and Biotechnological ProductionPublication . Andrade, Mariana; Santos, Regiane; Sanches-silva, AnaEssential oils have been used since the discovery of fre by many civiliza tions. Alchemists used to produce the Quinta essentia by distillation, which now is known as essential oils. This review aims to provide a systematic overview of the composition in terms of active compounds and main biological activities of differ ent essential oils. In general terms, these include anti-infammatory, antidepressant, antioxidant, antitumor, antimicrobial, anticancer, and antimutagenic activities. In addition, the techniques to extract and evaluate their composition are dis cussed. Moreover, their main industrial applications, especially the application as favors and fragrances, in pharmaceutical and medicinal industry, in alternative medicines, in cosmetic industry, and in food industry are also reviewed and dis cussed in order to identify the future trends. Finally, the biotechnological produc tion of essential oils and their components was also assessed.
- Heat-related mortality in Portugal amplified during the COVID-19 pandemicPublication . Sousa, Pedro M.; Trigo, Ricardo; Russo, Ana; Geirinhas, João L.; Rodrigues, Ana Paula; Silva, Susana Pereira; Torres, Ana RitaAbout 2020 presented the highest mortality burden in recent history in Portugal (11% above the baseline)
- Nosocomial Outbreak of SARS-CoV-2 in a “Non-COVID-19” Hospital Ward: Virus Genome Sequencing as a Key Tool to Understand Cryptic TransmissionPublication . Borges, Vítor; Isidro, Joana; Macedo, Filipe; Neves, José; Silva, Luís; Paiva, Mário; Barata, José; Catarino, Judite; Ciobanu, Liliana; Duarte, Sílvia; Vieira, Luís; Guiomar, Raquel; Gomes, João PauloDissemination of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in healthcare institutions affects both patients and health-care workers (HCW), as well as the institutional capacity to provide essential health services. Here, we investigated an outbreak of SARS-CoV-2 in a "non-COVID-19" hospital ward unveiled by massive testing, which challenged the reconstruction of transmission chains. The contacts network during the 15-day period before the screening was investigated, and positive SARS-CoV-2 RNA samples were subjected to virus genome sequencing. Of the 245 tested individuals, 48 (21 patients and 27 HCWs) tested positive for SARS-CoV-2. HCWs were mostly asymptomatic, but the mortality among patients reached 57.1% (12/21). Phylogenetic reconstruction revealed that all cases were part of the same transmission chain. By combining contact tracing and genomic data, including analysis of emerging minor variants, we unveiled a scenario of silent SARS-CoV-2 dissemination, mostly driven by the close contact within the HCWs group and between HCWs and patients. This investigation triggered enhanced prevention and control measures, leading to more timely detection and containment of novel outbreaks. This study shows the benefit of combining genomic and epidemiological data for disclosing complex nosocomial outbreaks, and provides valuable data to prevent transmission of COVID-19 in healthcare facilities.
- Primary and oxidative DNA damage induced by titanium dioxide nanoparticles in salivary leukocytes: a non-invasive alternative in nanomaterials genotoxicity assessmentPublication . Fernández-Bertólez, Natalia; Rodríguez-Fernández, R.; Lema-Arranz, C.; Pásaro, E.; Reis, Ana Teresa; Teixeira, João Paulo; Costa, Carla; Laffon, Blanca; Valdiglesias, VanessaIntroduction Metal oxide nanoparticles (NP) have a wide variety of applications in consumer products and biomedical practices. Recently, salivary leucocytes have been proposed as non-invasive alternative to peripheral blood leucocytes to evaluate the toxic effects of recent exposure to environmental contaminants, particularly those involving inhalatory or oral exposure routes. The present study aimed at evaluating the potential genotoxicity of titanium dioxide (TiO2) NP, frequently present in consumer products, in salivary leucocytes in vitro. Primary and oxidative DNA damage were evaluated by the standard alkaline comet assay and the hOGG1 enzyme incubation modification, respectively. Possible interferences of the NP with the methodological procedure or the OGG1 activity were addressed prior to analyses.