Browsing by Issue Date, starting with "2021-09-07"
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- Comparison of ISO 6579–1, VIDAS Easy SLM, and SureFast® Salmonella ONE Real-time PCR, for Salmonella Detection in Different Groups of FoodstuffsPublication . Furtado, R.; Coelho, A.; Morais, M.; Leitão, A.L.; Saraiva, M.; Correia, C.B.; Batista, R.In the European Union (EU), Salmonella was the causative agent responsible for almost one in three (30.7%) of all foodborne outbreaks reported by member states during 2018, causing 11,581 cases of illness, which represented an increase of 20.6% compared to 2017. Considering the importance of this foodborne zoonotic bacterium in food safety and human health, several strategies for the control and consequent detection of Salmonella in foodstuffs are continuously being developed. In this study, we have tested 137 food samples (78 potentially naturally contaminated, 21 artificially contaminated with high levels of Salmonella, and 38 artificially contaminated with low levels of Salmonella) in order to compare the results and performance of three Salmonella detection methods: standard conventional culture (ISO 6579–1), SureFast® Salmonella ONE real-time PCR, and VIDAS® (Vitek Immunodiagnostic Assay System) Easy SLM, an Enzyme Linked Fluorescent Assay (ELFA). Although SureFast® Salmonella ONE real-time PCR was the fastest, it showed more inconclusive results, due to PCR inhibition and false positive results. ISO and VIDAS® protocols gave identical results and proved to be more robust than SureFast® Salmonella ONE real-time PCR when testing different food matrices, despite its longer response times. SureFast® Salmonella ONE real-time PCR may be appropriate to be used when the objective is to test food matrices that are known not to interfere with PCR and expected to be negative for Salmonella. All the analytical tested methods have advantages and limitations and thus, depending on the situation, may be used as the elected method for Salmonella detection in foodstuffs in accordance with the purpose of the laboratorial analysis.
- Global perspective of familial hypercholesterolaemia: a cross-sectional study from the EAS Familial Hypercholesterolaemia Studies Collaboration (FHSC)Publication . EAS Familial Hypercholesterolaemia Studies Collaboration (FHSC)Background: The European Atherosclerosis Society Familial Hypercholesterolaemia Studies Collaboration (FHSC) global registry provides a platform for the global surveillance of familial hypercholesterolaemia through harmonisation and pooling of multinational data. In this study, we aimed to characterise the adult population with heterozygous familial hypercholesterolaemia and described how it is detected and managed globally. Methods: Using FHSC global registry data, we did a cross-sectional assessment of adults (aged 18 years or older) with a clinical or genetic diagnosis of probable or definite heterozygous familial hypercholesterolaemia at the time they were entered into the registries. Data were assessed overall and by WHO regions, sex, and index versus non-index cases. Findings: Of the 61 612 individuals in the registry, 42 167 adults (21 999 [53·6%] women) from 56 countries were included in the study. Of these, 31 798 (75·4%) were diagnosed with the Dutch Lipid Clinic Network criteria, and 35 490 (84·2%) were from the WHO region of Europe. Median age of participants at entry in the registry was 46·2 years (IQR 34·3-58·0); median age at diagnosis of familial hypercholesterolaemia was 44·4 years (32·5-56·5), with 40·2% of participants younger than 40 years when diagnosed. Prevalence of cardiovascular risk factors increased progressively with age and varied by WHO region. Prevalence of coronary disease was 17·4% (2·1% for stroke and 5·2% for peripheral artery disease), increasing with concentrations of untreated LDL cholesterol, and was about two times lower in women than in men. Among patients receiving lipid-lowering medications, 16 803 (81·1%) were receiving statins and 3691 (21·2%) were on combination therapy, with greater use of more potent lipid-lowering medication in men than in women. Median LDL cholesterol was 5·43 mmol/L (IQR 4·32-6·72) among patients not taking lipid-lowering medications and 4·23 mmol/L (3·20-5·66) among those taking them. Among patients taking lipid-lowering medications, 2·7% had LDL cholesterol lower than 1·8 mmol/L; the use of combination therapy, particularly with three drugs and with proprotein convertase subtilisin-kexin type 9 inhibitors, was associated with a higher proportion and greater odds of having LDL cholesterol lower than 1·8 mmol/L. Compared with index cases, patients who were non-index cases were younger, with lower LDL cholesterol and lower prevalence of cardiovascular risk factors and cardiovascular diseases (all p<0·001). Interpretation: Familial hypercholesterolaemia is diagnosed late. Guideline-recommended LDL cholesterol concentrations are infrequently achieved with single-drug therapy. Cardiovascular risk factors and presence of coronary disease were lower among non-index cases, who were diagnosed earlier. Earlier detection and greater use of combination therapies are required to reduce the global burden of familial hypercholesterolaemia.
- Unraveling the role of internal ribosome entry site (IRES) mediated translation in human UPF1 expression and functionPublication . Lacerda, Rafaela; Menezes, Juliane; Romão, LuísaUp-frameshift 1 (UPF1) is the key factor in nonsense-mediated mRNA decay (NMD, a mechanism that degrades transcripts carrying premature translation termination codons. Besides its role in NMD, UPF1 has also a role in other mechanisms, such as cell cycle progression, being crucial for the G1/S transition, and in telomere maintenance and homeostasis. Furthermore, it has also been identified as having tumour suppressor activity in different cancers, such as hepatocellular carcinoma. Our data supports the existence of an IRES within the human UPF1 5’ untranslated region (UTR). In fact, using a bicistronic reporter system, we observed that UPF1 5’UTR can mediate internal translation in a 5’ cap-independent manner, both in normal and under stress conditions. We concluded that the first 100 nucleotides and the last 125 (out of a total of 275) are the minimal sequences essential for the identified IRES activity. According to the in silico predicted secondary structure, these two segments correspond to two stem-loops. Now, we are experimentally confirming this predicted secondary structure, and understanding the physiological role of this IRES-mediated activity in the NMD process. Also, we are testing its importance for UPF1 function as a tumour suppressor. For that, we are using custom-made antisense oligonucleotides that specifically target those stem-loops and see to what extent they inhibit UPF1 IRES-mediated activity and how such inhibition will play a role in the cell and the functions in which UPF1 participates. Also, we are testing some hallmarks of cancer, such as proliferation, invasion, and apoptosis inhibition when the UPF1 IRES is impaired. With this work, we wish to unravel a new layer of knowledge in gene expression regulation of UPF1.