experimental study

O ácido hialurónico é um glicosaminoglicano com capacidade de absorção de água, cuja utilização tem sido cada vez mais frequente em fillers injetáveis, de modo a melhorar as características estéticas da face. Com o crescente aumento da popularidade e utilização de fillers de ácido hialurónico na medicina estética, surgiu a necessidade de encontrar um antídoto eficaz para tratar possíveis complicações relacionadas com estes procedimentos, como por exemplo oclusões vasculares ou reações alérgicas. Neste contexto, a hialuronidase, uma enzima com capacidade de degradar o ácido hialurónico em ácido glucorónico e N-acetilglucosamina (NAG), tem sido amplamente utilizada. Esta enzima tem sido objeto de estudo crescente desde a primeira metade do século XX. Para além do seu uso na medicina estética, embora ainda off- label, é utilizado em diversas outras áreas da medicina. Em 1971, esta enzima foi bem documentada e classificada por Karl Meyer em três origens distintas: animal, microbiana e de moluscos, com base não só na sua estrutura como também na sua atividade enzimática. Após vários estudos, foi desenvolvida uma enzima com maior atividade, criada por engenharia recombinante, com recurso ao DNA de hamsters. Atualmente, as formulações mais frequentemente utilizadas na prática clínica são de origem animal e de origem recombinante. Apesar da importância da hialuronidase no contexto dos fillers de ácido hialurónico, existe uma escassez de estudos na literatura relativamente a este tema, nomeadamente na comparação da efetividade de hialuronidases de diferentes origens na degradação de ácido hialurónico. Além disso, a influência que as propriedades físicas dos fillers podem ter nesta degradação também necessita de maior investigação, apesar de ser um tema já mais desenvolvido. Não existe consenso no que diz respeito às concentrações de enzima a utilizar, sendo premente uma padronização para facilitar a aplicabilidade clínica destas enzimas. Com base nestas limitações, foi identificada a necessidade de realizar um estudo que comparasse a efetividade de duas hialuronidases de diferentes origens na degradação de fillers injetáveis de ácido hialurónico, tendo sido esse o objetivo do estudo experimental apresentado. Para a realização do estudo experimental, foram utilizados dois fillers de ácido hialurónico da marca Fillmed® (França) e duas hialuronidases distintas, uma animal da marca InstitutoBcn® (Espanha) e outra recombinante da pbserum® (Espanha), formando assim quatro grupos de estudo: ART FILLER® Universal com hialuronidase animal, ART FILLER® Universal com hialuronidase recombinante, ART FILLER® Volume com hialuronidase animal e ART FILLER® Volume com hialuronidase recombinante. A seleção de fillers e hialuronidases para este estudo baseou-se no que é frequentemente utilizado em Portugal e nas opções às quais tínhamos acesso mais facilmente. A marca Fillmed® é apresentada em poucos estudos na literatura, daí haver uma necessidade acrescida do seu estudo. A nível de hialuronidases, optou- se por utilizar uma hialuronidase de origem animal devido ao extenso histórico de estudos a que esta já foi submetida ao longo dos anos, e uma hialuronidase de origem recombinante, por ser uma formulação mais recente e com boas propriedades já documentadas na literatura. A concentração utilizada de cada hialuronidase foi de 75 UI (Unidades Internacionais) para cada 0,1ml de cada filler (25mg/ml), o que vai de encontro à literatura previamente estudada, dentro das limitações que esta apresenta. Para chegar a esta concentração, ambas as hialuronidases foram diluídas em solução salina. A cada grupo foi adicionado um reagente, p- dimetilaminobenzaldeído, também conhecido como reagente de Ehrlich, que tem a capacidade de reagir com as cadeias de NAG, libertadas pelo ácido hialurónico aquando da sua degradação por hialuronidase, fornecendo uma cor violeta à amostra. Utilizando uma técnica colorimétrica, a quantidade de NAG libertada foi posteriormente medida através da absorbância (A) com recurso a um medidor de placas (Perkin Elmer® Inc., USA). Quanto maior a degradação do ácido hialurónico, mais cadeias de NAG livres, e assim, maior será o valor de absorbância, uma vez que haverá uma cor mais intensa do reagente. Esta medição da absorbância foi realizada em quatro momentos distintos ao longo do tempo: após 1, 6, 24 e 48 horas de incubação a 37ºC, de modo mimetizar as condições de temperatura no corpo humano, com N=5. Os resultados de absorbância foram apresentados sobre a forma de média ± desvio padrão (DP) e posteriormente analisados recorrendo a um teste não-paramétrico, após se mostrar a ausência de normalidade da amostra, através de um software específico de análise estatística. H0: não existem diferenças na degradação dos dois fillers pelas duas hialuronidases. A utilização rigorosa deste protocolo experimental permitiu chegar a resultados significativos, onde se demonstrou que ambos os fillers sofreram degradação enzimática por ambas as hialuronidases (animal e recombinante). Após a análise estatística dos resultados, com recurso a um teste não-paramétrico, a hipótese nula foi rejeitada, provando que existem diferenças estatisticamente significativas na degradação dos fillers pelas duas hialuronidases. No entanto, ao realizar uma comparação das duas hialuronidases individualmente na degradação de cada filler, compreendeu-se que só se verificaram diferenças estatisticamente significativas na degradação de ART FILLER® Universal pelas hialuronidases às 24 horas (p=0,007), tempo no qual a hialuronidase recombinante se mostrou mais efetiva em comparação com a animal. No entanto, ao analisar o comportamento de ART FILLER® Volume ao longo do tempo, constatamos uma degradação estatisticamente significativa maior pela hialuronidase recombinante às 6 horas (p<0,001), 24 horas (p=0,007) e 48 horas (p<0,001) quando comparado com a hialuronidase animal. Estes dados mostram que a hialuronidase recombinante apresenta maior degradação de ART FILLER® Volume logo depois das 6h. Ao realizar a comparação individual dos dois fillers, quando submetidos à mesma hialuronidase, apenas se observaram diferenças estatisticamente significativas à 1h, na degradação destes por hialuronidase animal (p<0,001), onde se observou uma maior degradação de ART FILLER® Universal. A nível de degradação pela hialuronidase recombinante, não foram demonstradas diferenças entre os fillers. Assim, observou-se que não existem diferenças significativas no filler em si que possam influenciar a sua degradação, quando comparado com o outro filler em análise, após 6h de incubação. Com base nos resultados obtidos, o nosso estudo permitiu concluir que, entre todos os grupos analisados, a hialuronidase que apresenta melhores resultados ao longo do tempo é a recombinante, principalmente na degradação de ART FILLER® Volume, enquanto ART FILLER® Universal não mostrou diferenças significativas na sua degradação pelas duas enzimas na maioria dos tempos medidos. Também se observou que não há diferenças na degradação dos dois fillers diferentes quando submetidos à mesma hialuronidase, nem quando submetidos à de origem animal nem à de origem recombinante, a partir das 6h de incubação. O estudo realizado contribui para a evolução do conhecimento e da literatura existente, porém, é importante destacar que existiram diversas limitações que podem ser superadas com a formulação de estudos futuros na área. Entre essas limitações encontram-se a falta de recursos, o que impossibilitou a realização de medições mais frequentes ao longo do tempo, para uma melhor compreensão do comportamento da enzima e da sua interação com o ácido hialurónico. Para além disso, tratando-se de um estudo in vitro há sempre condições que são difíceis de mimetizar e deve-se ter isso em consideração ao extrapolar os resultados deste estudo para a prática clínica. Muitos dos estudos presentes na literatura utilizam concentrações de hialuronidase muito elevadas, que dificilmente são extrapoladas para a prática clínica sem nenhum risco acrescido. Sugerimos que futuras pesquisas avaliem não só as duas hialuronidases mencionadas neste estudo como também analisem o seu comportamento ao longo de um maior período de tempo e com diferentes concentrações.Objectives: This study aimed to evaluate and compare the effectiveness of two hyaluronidases in degrading two different hyaluronic acid fillers. Materials and methods: Four groups were tested: ART FILLER® Universal and animal hyaluronidase, ART FILLER® Universal and recombinant hyaluronidase, ART FILLER® Volume and animal hyaluronidase and ART FILLER® Volume and recombinant hyaluronidase (N=5). A reagent that reacts with N-acetylglucosamine (NAG) was added. Absorbance was measured at 585 nm after 1, 6, 24 and 48 hours of incubation at 37ºC. Results were expressed as mean and standard deviation (±SD) of absorbance and analyzed using a non-parametric test. Results: All tested HA fillers were susceptible to degradation by hyaluronidase. Comparing the degradation of ART FILLER® Universal by the two hyaluronidases, statistically significant differences were observed at 24 hours, with recombinant hyaluronidase showing higher degradation than animal (p=0,007). For ART FILLER® Volume, recombinant hyaluronidase resulted in higher absorbance values than animal, throughout the entire time of degradation, except at 1h (6h p<0,001; 24h p=0,007; 48h p<0,001). Differences were also observed when comparing the effect of the same hyaluronidase on degrading both HA fillers. Animal hyaluronidase demonstrated greater effectiveness in degrading ART FILLER® Universal, compared to ART FILLER® Volume, particularly significant at 1h (p<0,001). However, no statistically differences in the degradation of the two HA fillers were found with recombinant hyaluronidase. Conclusion: Recombinant hyaluronidase appeared to be the most effective overall. However, the significantly higher degradation values compared to animal hyaluronidase were only observed when paired with ART FILLER® Volume, at 6, 24 and 48 hours. Therefore, in clinical practice, recombinant hyaluronidase should be preferred when treating complications associated with prior injection of ART FILLER® Volume. When comparing the two HA fillers, no significant differences were observed, except at 1h, when animal hyaluronidase exhibited greater effectiveness in degrading ART FILLER® Universal

Score CTo-aBCDE : um novo score preditor de sucesso nas CTOs

© 2020 Sociedade Portuguesa de Cardiologia. Published by Elsevier España, S.L.U. This is an open access article under the CC-BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).Introduction: Patient selection for percutaneous coronary intervention (PCI) in chronic total occlusions (CTOs) is crucial to procedural success. Our aim was to identify independent predictors of success in CTO PCI in order to create an accurate score. Methods: In a single-center observational registry of CTO PCI, demographic and clinical data and anatomical characteristics of coronary lesions were recorded. Linear and logistic regression analysis were used to identify predictors of success. A score to predict success was created and its accuracy was measured by receiver operating curve analysis. Results: A total of 377 interventions were performed (334 patients, age 68±11 years, 75% male). The success rate was 65% per patient and 60% per procedure. Predictors of success in univariate analysis were absence of active smoking (OR 2.02, 95% CI 1.243-3.29; p=0.005), presence of tapered stump (OR 5.2, 95% CI 2.7-10.2; p8 with high probability (95%). Conclusion: In our sample only anatomical characteristics were predictors of success. The creation of a score to predict success, with good accuracy, may enable selection of cases that can be treated by any operator, those in which a dedicated operator will be desirable, and those with an extremely low probability of success, which should be considered individually for conservative management, surgical revascularization or PCI by a team experienced in CTO.info:eu-repo/semantics/publishedVersio

Identification of clusters of asthma control: A preliminary analysis of the inspirers studies

This work was funded by ERDF (European Regional Development Fund) through the operations: POCI- -01-0145-FEDER-029130 (“mINSPIRERS—mHealth to measure and improve adherence to medication in chronic obstructive respiratory diseases - generalisation and evaluation of gamification, peer support and advanced image processing technologies”) co-funded by the COMPETE2020 (Programa Operacional Competitividade e Internacionalização), Portugal 2020 and by Portuguese Funds through FCT (Fundação para a Ciência e a Tecnologia).© 2020, Sociedade Portuguesa de Alergologia e Imunologia Clinica. All rights reserved. Aims: To identify distinct asthma control clusters based on Control of Allergic Rhinitis and Asthma Test (CARAT) and to compare patients’ characteristics among these clusters. Methods: Adults and adolescents (≥13 years) with persistent asthma were recruited at 29 Portuguese hospital outpatient clinics, in the context of two observational studies of the INSPIRERS project. Demographic and clinical characteristics, adherence to inhaled medication, beliefs about inhaled medication, anxiety and depression, quality of life, and asthma control (CARAT, >24 good control) were collected. Hierarchical cluster analysis was performed using CARAT total score (CARAT-T). Results: 410 patients (68% adults), with a median (percentile 25–percentile 75) age of 28 (16-46) years, were analysed. Three clusters were identified [mean CARAT-T (min-max)]: cluster 1 [27(24-30)], cluster 2 [19(14-23)] and cluster 3 [10(2-13)]. Patients in cluster 1 (34%) were characterised by better asthma control, better quality of life, higher inhaler adherence and use of a single inhaler. Patients in clusters 2 (50%) and 3 (16%) had uncontrolled asthma, lower inhaler adherence, more symptoms of anxiety and depression and more than half had at least one exacerbation in the previous year. Further-more, patients in cluster 3 were predominantly female, had more unscheduled medical visits and more anxiety symp-toms, perceived a higher necessity of their prescribed inhalers but also higher levels of concern about taking these inhalers. There were no differences in age, body mass index, lung function, smoking status, hospital admissions or specialist physician follow-up time among the three clusters. Conclusion: An unsupervised method based on CARAT--T, identified 3 clusters of patients with distinct, clinically meaningful characteristics. The cluster with better asthma control had a cut-off similar to the established in the validation study of CARAT and an additional cut-off seems to distinguish more severe disease. Further research is necessary to validate the asthma control clusters identified.publishersversionpublishe

A small TAT-TrkB peptide prevents BDNF receptor cleavage and restores synaptic physiology in Alzheimer's disease

Publisher Copyright: Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.In Alzheimer's disease (AD), amyloid β (Aβ)-triggered cleavage of TrkB-FL impairs brain-derived neurotrophic factor (BDNF) signaling, thereby compromising neuronal survival, differentiation, and synaptic transmission and plasticity. Using cerebrospinal fluid and postmortem human brain samples, we show that TrkB-FL cleavage occurs from the early stages of the disease and increases as a function of pathology severity. To explore the therapeutic potential of this disease mechanism, we designed small TAT-fused peptides and screened their ability to prevent TrkB-FL receptor cleavage. Among these, a TAT-TrkB peptide with a lysine-lysine linker prevented TrkB-FL cleavage both in vitro and in vivo and rescued synaptic deficits induced by oligomeric Aβ in hippocampal slices. Furthermore, this TAT-TrkB peptide improved the cognitive performance, ameliorated synaptic plasticity deficits and prevented Tau pathology progression in vivo in the 5XFAD mouse model of AD. No evidence of liver or kidney toxicity was found. We provide proof-of-concept evidence for the efficacy and safety of this therapeutic strategy and anticipate that this TAT-TrkB peptide has the potential to be a disease-modifying drug that can prevent and/or reverse cognitive deficits in patients with AD.publishersversionpublishe

Determinants of HIV late presentation among men who have sex with men in Portugal (2014–2019): who’s being left behind?

Introduction: HIV late presentation (LP) remains excessive in Europe. We aimed to analyze the factors associated with late presentation in the MSM population newly diagnosed with HIV in Portugal between 2014 and 2019. Methods: We included 391 newly HIV-1 diagnosed Men who have Sex with Men (MSM), from the BESTHOPE project, in 17 countrywide Portuguese hospitals. The data included clinical and socio-behavioral questionnaires and the viral genomic sequence obtained in the drug resistance test before starting antiretrovirals (ARVs). HIV-1 subtypes and epidemiological surveillance mutations were determined using different bioinformatics tools. Logistic regression was used to estimate the association between predictor variables and late presentation (LP). Results: The median age was 31 years, 51% had a current income between 501-1,000 euros, 28% were migrants. 21% had never been tested for HIV before diagnosis, with 42.3% of MSM presenting LP. 60% were infected with subtype B strains. In the multivariate regression, increased age at diagnosis, higher income, lower frequency of screening, STI ever diagnosed and higher viral load were associated with LP. Conclusion: Our study suggests that specific subgroups of the MSM population, such older MSM, with higher income and lower HIV testing frequency, are not being targeted by community and clinical screening services. Overall, targeted public health measures should be strengthened toward these subgroups, through strengthened primary care testing, expanded access to PrEP, information and promotion of HIV self-testing and more inclusive and accessible health services.info:eu-repo/semantics/publishedVersio

SARS-CoV-2 introductions and early dynamics of the epidemic in Portugal

Genomic surveillance of SARS-CoV-2 in Portugal was rapidly implemented by the National Institute of Health in the early stages of the COVID-19 epidemic, in collaboration with more than 50 laboratories distributed nationwide. Methods By applying recent phylodynamic models that allow integration of individual-based travel history, we reconstructed and characterized the spatio-temporal dynamics of SARSCoV-2 introductions and early dissemination in Portugal. Results We detected at least 277 independent SARS-CoV-2 introductions, mostly from European countries (namely the United Kingdom, Spain, France, Italy, and Switzerland), which were consistent with the countries with the highest connectivity with Portugal. Although most introductions were estimated to have occurred during early March 2020, it is likely that SARS-CoV-2 was silently circulating in Portugal throughout February, before the first cases were confirmed. Conclusions Here we conclude that the earlier implementation of measures could have minimized the number of introductions and subsequent virus expansion in Portugal. This study lays the foundation for genomic epidemiology of SARS-CoV-2 in Portugal, and highlights the need for systematic and geographically-representative genomic surveillance.We gratefully acknowledge to Sara Hill and Nuno Faria (University of Oxford) and Joshua Quick and Nick Loman (University of Birmingham) for kindly providing us with the initial sets of Artic Network primers for NGS; Rafael Mamede (MRamirez team, IMM, Lisbon) for developing and sharing a bioinformatics script for sequence curation (https://github.com/rfm-targa/BioinfUtils); Philippe Lemey (KU Leuven) for providing guidance on the implementation of the phylodynamic models; Joshua L. Cherry (National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health) for providing guidance with the subsampling strategies; and all authors, originating and submitting laboratories who have contributed genome data on GISAID (https://www.gisaid.org/) on which part of this research is based. The opinions expressed in this article are those of the authors and do not reflect the view of the National Institutes of Health, the Department of Health and Human Services, or the United States government. This study is co-funded by Fundação para a Ciência e Tecnologia and Agência de Investigação Clínica e Inovação Biomédica (234_596874175) on behalf of the Research 4 COVID-19 call. Some infrastructural resources used in this study come from the GenomePT project (POCI-01-0145-FEDER-022184), supported by COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation (POCI), Lisboa Portugal Regional Operational Programme (Lisboa2020), Algarve Portugal Regional Operational Programme (CRESC Algarve2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), and by Fundação para a Ciência e a Tecnologia (FCT).info:eu-repo/semantics/publishedVersio

HIV-1-Transmitted Drug Resistance and Transmission Clusters in Newly Diagnosed Patients in Portugal Between 2014 and 2019

Objective: To describe and analyze transmitted drug resistance (TDR) between 2014 and 2019 in newly infected patients with HIV-1 in Portugal and to characterize its transmission networks. Methods: Clinical, socioepidemiological, and risk behavior data were collected from 820 newly diagnosed patients in Portugal between September 2014 and December 2019. The sequences obtained from drug resistance testing were used for subtyping, TDR determination, and transmission cluster (TC) analyses. Results: In Portugal, the overall prevalence of TDR between 2014 and 2019 was 11.0%. TDR presented a decreasing trend from 16.7% in 2014 to 9.2% in 2016 (p for-trend = 0.114). Multivariate analysis indicated that TDR was significantly associated with transmission route (MSM presented a lower probability of presenting TDR when compared to heterosexual contact) and with subtype (subtype C presented significantly more TDR when compared to subtype B). TC analysis corroborated that the heterosexual risk group presented a higher proportion of TDR in TCs when compared to MSMs. Among subtype A1, TDR reached 16.6% in heterosexuals, followed by 14.2% in patients infected with subtype B and 9.4% in patients infected with subtype G. Conclusion: Our molecular epidemiology approach indicates that the HIV-1 epidemic in Portugal is changing among risk group populations, with heterosexuals showing increasing levels of HIV-1 transmission and TDR. Prevention measures for this subpopulation should be reinforced.info:eu-repo/semantics/publishedVersio

Trajetórias da Educomunicação nas Políticas Públicas e a Formação de seus Profissionais

Esta obra é composta com os trabalhos apresentados no primeiro subtema, TRAJETÓRIA – Educação para a Comunicação como Política pública, nas perspectivas da Educomunicação e da Mídia-Educação, do II Congresso Internacional de Comunicação e Educação. Os artigos pretendem propiciar trocas de informações e produzir reflexões com os leitores sobre os caminhos percorridos, e ainda a percorrer, tendo como meta a expansão e a legitimação das práticas educomunicativas e/ou mídia-educativas como política pública para o atendimento à formação de crianças, adolescentes, jovens e adultos, no Brasil e no mundo

COVID-19 symptoms at hospital admission vary with age and sex: results from the ISARIC prospective multinational observational study

Background: The ISARIC prospective multinational observational study is the largest cohort of hospitalized patients with COVID-19. We present relationships of age, sex, and nationality to presenting symptoms. Methods: International, prospective observational study of 60 109 hospitalized symptomatic patients with laboratory-confirmed COVID-19 recruited from 43 countries between 30 January and 3 August 2020. Logistic regression was performed to evaluate relationships of age and sex to published COVID-19 case definitions and the most commonly reported symptoms. Results: ‘Typical’ symptoms of fever (69%), cough (68%) and shortness of breath (66%) were the most commonly reported. 92% of patients experienced at least one of these. Prevalence of typical symptoms was greatest in 30- to 60-year-olds (respectively 80, 79, 69%; at least one 95%). They were reported less frequently in children (≤ 18 years: 69, 48, 23; 85%), older adults (≥ 70 years: 61, 62, 65; 90%), and women (66, 66, 64; 90%; vs. men 71, 70, 67; 93%, each P &lt; 0.001). The most common atypical presentations under 60 years of age were nausea and vomiting and abdominal pain, and over 60 years was confusion. Regression models showed significant differences in symptoms with sex, age and country. Interpretation: This international collaboration has allowed us to report reliable symptom data from the largest cohort of patients admitted to hospital with COVID-19. Adults over 60 and children admitted to hospital with COVID-19 are less likely to present with typical symptoms. Nausea and vomiting are common atypical presentations under 30 years. Confusion is a frequent atypical presentation of COVID-19 in adults over 60 years. Women are less likely to experience typical symptoms than men