A catalogue of stress granules´ components: implications for neurodegeneration

Stress granules (SGs) are irregularly shaped foci constituted by a variety of different types of RNAs, proteins, factors involved in translation and signaling molecules. These granules form transiently in response to a variety of stress stimuli, such as viral infections or translation blocking drugs, and facilitate stress response by sequestering mRNAs and proteins and affecting the translation of RNA transcripts. SGs are necessary for stress response, however, abnormalities in SGs functioning and the association of their formation with the aggregation of many pathological proteins, have been linked to pathological changes in several human diseases, such as neurodegenerative diseases, cancer, and aging. The aim of this work was to identify all the proteins described as SGs components in mammalian cells and to build an online open access database with the all information collected. The SGs components were identified through an exhaustive study search (PubMed) and further details about each component were retrieved using public databases. Moreover, a transcriptomic analysis was performed analyzing the gene expression data for each SGs component in different neurodegenerative diseases. We identified 464 proteins as components of mammalian cells SGs, from which 253 were classified as RNA-binding proteins (RBPs). Details about each protein, such as molecular function, link to disease, cell type where they were detected and the stress stimuli used to induce SGs assembly, were also collected and are available in the database. Through a transcriptomic analysis, where “disease vs control” groups were compared and analyzed, a vast majority of SGs proteins were found to be differentially expressed in different neurodegenerative diseases. All the information collected was used to build the Mammalian Stress Granules Proteome (MSGP), available at https://msgp.pt/, an important tool for researchers in this area of growing interest, being the first database to list and provide information about all SGs proteins identified so far.Em condições de stresse, os organismos eucariotas possuem um conjunto de mecanismos para o combater e assim restabelecer a homeostasia celular. Nesta resposta inclui-se a formação dos grânulos de stresse, que são estruturas de formato irregular, sem membrana e formados de forma transiente, que sequestram mRNAs e proteínas, afectando a tradução, e assim facilitando a resposta celular ao stresse. Os grânulos de stresse são constituidos por uma grande variedade de componentes: proteínas, mRNAs e microRNAs, componentes da tradução e moléculas de sinalização. No entanto, o perfil proteíco destes grânulos varia consoante o tipo celular onde se formam e o tipo de stresse celular, como por exemplo, a privação nutricional, infecções virais, radiações UV, entre outros. Para a formação dos grânulos de stresse são necessárias proteínas ligantes a RNA (RNA-binding proteins – RBPs). As RBPs são proteínas envolvidas numa variedade de processos relacionados com RNA, como o splicing alternativo e a biogénese de miRNA, possuindo um papel muito importante na função neuronal. Estas proteínas, para além de fazerem parte da constituição dos grânulos de stresse, são também fundamentais para a sua formação. Tal como já foi referido a formação dos grânulos de stresse é temporária, uma vez que quando o stresse desaparece, estes grânulos são eliminados. Esta eliminação é feita através de autofagia, mais especificamente por macroautofagia, na qual a proteína Valosin-containing protein (VCP) é fundamental, ou por autofagia mediada por chaperonas. Embora por um lado os grânulos de stress sejam essenciais na resposta da célula ao stresse, por outro lado, defeitos na sua formação, dinâmica ou eliminação podem contribuir para a patogénese de diversas doenças humanas, como por exemplo, cancro, e doenças neurodegenerativas. De facto, existem cada vez mais evidências que ligam os grânulos de stresse a doenças neurodegenerativas, nomeadamente as doenças de Alzheimer, Parkinson, Huntigton ou Esclerose Lateral Amiotrófica. Para além disso, várias das proteínas envolvidas na patogénese destas doenças também se encontram presentes nos grânulos de stress, como é o caso da Atx-2, Tau, FUS ou TDP-43. A importante função dos grânulos de stresse a nível celular e a sua implicação em diversas patologias torna-os alvos importantes para o estudos dessas doenças e até como possíveis alvos terapêuticos. O principal objetivo deste trabalho foi identificar todos os componentes proteícos dos grânulos de stresse descritos em células de mamífero, recolher informações sobre os mesmos e catalogar toda a informação recolhida numa base de dados online. Os componentes proteícos dos grânulos de stresse foram identificados através duma pesquisa exaustiva nos estudos disponíveis no PubMed, tendo sido filtradas apenas as proteínas existentes em células de mamífero. Para cada proteína, informações como o nome completo da proteína, gene ID, UniProt ID, se é ou não uma RBP, função molecular ou se está envolvida nalguma doença, foram obtidas através de bases de dados públicas. Para além disso, também foram recolhidas informações acerca do tipo celular em que as proteínas foram estudadas e tipo de estímulo utilizado para induzir o stress. Uma análise transcriptómica foi efetuada, utilizando dados de estudos que analisaram a expressão génica em indíviduos com as doenças de Alzheimer, Parkinson, Huntington, Esclerose Lateral Amiotrófica e também em indíviduos não-doentes (controlos). Nesta análise os grupos de doença versus controlo foram comparados, de forma a investigar a expressão diferencial dos componentes dos grânulos de stress. Foram catalogadas 464 proteínas, das quais 253 foram identificadas como RBPs e 111 já tinham sido previamente ligadas a doenças. Para além disso, 32 proteínas foram identificadas como estando relacionadas com a autofagia. Do total das 464 proteínas identificadas, 225 têm como função molecular a ligação, 308 encontram-se envolvidas em processos celulares e 131 pertencem à classe proteíca de ligação a ácidos nucleícos. Os dados da análise da expressão génica entre os grupos “doença” e “controlo”, demonstraram que a grande maioria dos componentes dos grânulos de stress encontram-se com a expressão alterada nas doenças neurodegenerativas estudadas. Por exemplo, verificou-se que na doença de Alzheimer, 187 componentes tinham a sua expressão significativamente aumentada, enquanto que 193 tinham a sua expressão diminuída. O perfil proteíco dos grânulos de stresse varia consoante o tipo celular ou tipo de estímulo de stress induzido. Assim, foi possível observar que a maioria das proteínas, mais especificamente 327, foram identificadas nas células U-2OS, enquanto 427 foram identificadas em estudos que utilizaram o arsenito de sódio como estímulo Toda a informação recolhida foi catalogada na base de dados Mammalian Stress Granules Proteome (MSGP), disponível para todos os investigadores em https://msgp.pt/. A base de dados MSGP possui a listagem de todos os componentes proteícos dos grânulos de streses assim como informações dos mesmos e dados de expressão génica para cada uma dos componentes identificados. O utilizador pode pesquisar por uma proteína específica utilizando a “search bar” ou pode filtrar a pesquisa utilizando as diferentes categorias e tags disponíveis. A MSGP é a primeira base de dados que reune todos os componentes dos grânulos descritos, constituindo uma importante ferramenta para os investigadores desta área, tendo para além disso, a possibilidade de continuar em crescimento, através da adição de informações não só acerca dos componentes já catalogados, como de outros componentes dos grânulos de stress, como por exemplo microRNAs

Gestão materna do mundo familiar e laboral em famílias nucleares com crianças em idade pré-escolar. Associações com o envolvimento paterno

A entrada da mulher no mercado de trabalho provocou alterações na organização e nas expectativas acerca dos papéis parentais das famílias portuguesas. Surgiram, por isso, novos papéis parentais e um novo modelo familiar de coparentalidade, em que os pais e mães devem dividir tarefas e responsabilidades igualitariamente no domínio familiar. A ampliação do papel do pai e o seu envolvimento foi fulcral para ajudar a mãe. Este estudo visou analisar a gestão dos contextos trabalho-família (os benefícios e constrangimentos) na perspetiva da mãe, e as possíveis associações com o grau e tipo de envolvimento do pai nas atividades de cuidado e socialização da criança. Para tal, foram aplicados dois questionários sobre a conciliação dos contextos familiar e laboral e sobre o envolvimento parental, a 90 mães portuguesas. Os resultados indicam que os benefícios e constrangimentos da gestão materna dos contextos familiar e laboral, nesta amostra, não se encontram relacionados, com o envolvimento do pai. O número de horas de trabalho dos pais encontra-se correlacionado com os constrangimentos na gestão dos contextos laboral e familiar e influenciam a ocorrência de interferências entre contextos, bem como a participação nas dimensões de cuidados indiretos e de ensino e disciplina do envolvimento parental. Apesar disso, a gestão materna destes contextos tende a ser positiva para a mesma.The entry of women into the labor market has led to changes in the organization and expectations about parental roles of Portuguese families. Therefore, new parental roles and a new family model of co-parenting, in which both parents should equally share tasks and responsibilities in the family domain, have emerged. The expanded role of the father and his involvement was crucial to help the mother. This study examines the management of work-family contexts (both benefits and constraints) through the mother’s perspective, and the possible associations with the degree and type of the father’s involvement in the child’s care and socialization activities. For this purpose, two questionnaires were applied to 90 Portuguese mothers on the conciliation of family and work contexts, and on parental involvement. This sample’s results indicate that benefits and constraints of maternal management in work and family contexts are not related to the father’s involvement. The parents’ number of working hours is correlated with constraints in work and family context management and influence the occurrence of interference between contexts, as well as participation in the indirect care, teaching and parent involvement dimensions. Nevertheless, the management of these contexts tends to be beneficial for the mothe

Synthesis and characterization of electrical conductive bacterial cellulose-graft-polyaniline

Dissertação de mestrado em Micro-Nano TecnologiasIn recent years, conductive polymeric nanomaterials have received considerable attention because of the increasing demand of new technologies for the development of electronic devices, sensors, scaffolds, to be used in the most diverse research areas such as biology, tissue engineering, food industry, etc. In this context interest in polyaniline (PANI) has grown exponentially, being regarded as one of the most technologically capable electrical conducting polymers, due to its high electrical conductivity, easy synthesis, low cost, and stable electrical conductivity. Bacterial cellulose (BC) nanofibers appear as very promising support material for these conductive additives due to their high strength and stiffness associated to high purity, high porosity, and biocompatibility. The combination of these two materials opens a new field of potential applications for bacterial cellulose. This project aim was to develop an electrical conductive bacterial cellulose-graft-polyaniline composite by means of the oxidative-radical copolymerization using ammonium persulfate in acidic medium. The grafting conditions were studied by varying grafting parameters: monomer concentration and polymerization time. Different methods were studied for the producing of the conductive BC-graft-PANI composites: in situ direct polymerization of aniline in BC discs; surface modification of BC; and sulfonation of BC. The electrical conductivity increased from 7.5*10-11 S/cm to 2.26*10-4 S/cm by controlling the time of polymerization and the molar Ratio of CB:aniline. Under the assayed experimental conditions, the optimum grafting efficiency was find at a CB:aniline ratio of 1:10 and with a time reaction of 6 hours. The produced BC-graft-PANI composites were characterized using conductivity assays, scanning electron microscopy (SEM), fourier-transformed infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and viability assay (MTS test) taking BC as reference.Nos últimos anos, os polímeros condutores tem recebido especial atenção devido há crescente necessidade de novas tecnologias para o de envolvimento de dispositivos electrónicos, sensores, suportes porosos, que poderão ser usados nas mais diversas áreas como a biologia, a engenharia de tecidos, a industria alimentar, etc.. Neste contexto, o interesse na polianilina (PANI) tem crescido exponencialmente, sendo considerada um dos polímeros condutores mais promissores, devido à sua elevada condutividade eléctrica, fácil síntese, baixo custo e estabilidade térmica. No que diz respeito às nanofibras de celulose bacteriana, estas surgem como um material de suporte muito interessante para este tipo de polímeros condutores devido às excelentes propriedades mecânicas, associadas á elevada pureza, elevada porosidade, e biocompatibilidade. Com a combinação destes dois materiais abre-se um novo campo de aplicações para a celulose bacteriana. Este projecto tem como objectivo o desenvolvimento de um material compósito condutor à base de celulose bacteriana e polianilina através da polimerização oxidativa in situ da anilina em meio acido e usando persulfato de amónia como agente oxidante. As condições de polimerização foram estudadas fazendo variar os parâmetros de reacção: concentração do monómero e tempo de polimerização. Para a produção dos compósitos BC/PANI foram estudados diferentes métodos: a polimerização directa in situ da anilina na celulose bacteriana em meio ácido; modificação da superfície da celulose bacteriana, através da activação inicial da superfície da CB e posterior polimerização oxidativa; e sulfonação da celulose bacteriana. A condutividade eléctrica aumentou de 7.5*10-11 S/cm para 2.26*10-4 S/cm controlando o tempo de polimerização e a razão molar BC:anilina. Para as condições analisadas, os melhores resultados obtidos foram a razão molar CB:anilina de 1:10 e um tempo de reacção de 6 horas. Os materiais obtidos foram caracterizados através de ensaios de condutividade, microscopia electrónica de varrimento (SEM), espectroscopia de infravermelho (FTIR), analise termogravimétrica (TGA), calorimetria diferencial de varrimento (DSC), e ensaios de viabilidade (teste MTS), usando a celulose bacteriana como referência

Outcomes of lenvatinib therapy in poorly differentiated thyroid carcinoma

Funding Information: Ricardo Rodrigues was granted with a PhD scholarship by iNOVA4Health Research Unit (UIDP/04462/2020; UI/BD/154256/2022). The remaining authors did not receive any specific grant from funding agencies in the public, commercial, or non-for-profit sectors. Publisher Copyright: © 2023 the author(s) Published by Bioscientifica Ltd.Background and objective: Lenvatinib showed promising results in a subgroup of patients with poorly differentiated thyroid carcinoma (PDTC) in the SELECT trial. Our aim was to report the effectiveness and tolerability of lenvatinib in our series of PDTC patients. Methods: Medical records of eight consecutive patients with PDTC treated with lenvatinib in a single center between January 2019 and October 2022 were retrospectively reviewed. Inclusion criteria were PDTC diagnosis based on Turin criteria and evidence of disease progression in the previous 6 months. Results: Eight PDTC patients received an average dose of lenvatinib of 18.1 mg for a median duration of treatment of 10.3 months. The baseline Eastern Cooperative Oncology Group performance status was ≥2 in 50% of patients. Two patients had unresectable primary tumor. Seven patients showed extrathyroidal disease, particularly mediastinal lymph nodes (85.7%), lung (71.4%), and bone (71.4%). The disease control rate was 100%, with partial response and stable disease in 12.5 and 87.5%, respectively. The median time to best overall response was 3 months, and the median duration of response was 7.5 months. Median progression-free survival was 12 months and median overall survival was not reached. At 6, 12, and 18 months, overall survival was 87.5, 71.4, and 57.1%, respectively. All patients experienced drug-related adverse effects (AEs). Four (50%) had dose reductions and two (25%) had temporary treatment interruptions. Lenvatinib was stopped in two patients due to grade ≥3 AEs. Conclusion: Lenvatinib is an effective treatment for real-world PDTC patients. Adequate management of comorbidities and AEs increases treatment tolerability and minimizes dose reductions.publishersversionpublishe

Portuguese isolates of Aspergillus section Flavi unraveled by the calmodulin gene

Aspergillus is a large genus, with a complex and ever evolving taxonomy. Section Flavi is one of the most significant sections in the genus. Taxonomy and species identification is subject of great interest for scientists aiming to clarify the species concept and limits within the section. Furthermore, this section comprises both toxigenic and non-toxigenic species/strains, with great interest to biotechnology and food industry. Various genes, namely the rRNA (ITS region), calmodulin and β-tubulin genes, have been widely reported as good markers for Aspergillus species identification, because they are rapid and cost-effective. In the present study, we evaluated the discriminatory power of the ITS region and the calmodulin gene to distinguish closely related taxa within Aspergillus section Flavi. For this purpose, 26 isolates of Aspergillus section Flavi obtained from Portuguese almonds were characterized at various levels: i) phenotypic, regarding various aspects of morphology and physiology; ii) spectral, using MALDI-TOF ICMS to obtain protein fingerprinting; and iii) genotypic, by sequence analysis of a 730 bp segment of the calmodulin gene and a 908 bp segment of the ITS region. For the various methods, dendrograms were created and results were compared. Both genotypic and spectral analyses divided the isolates in 3 groups corresponding to closely related taxa of A. flavus, A. parasiticus and A. tamarii. Except for the ITS region, all sets of analysis positioned 5 of the 26 isolates in two unidentified clades close to A. parasiticus, and divided the A. flavus group in two distinct clades. The phylogenetic analysis of the calmodulin sequences resulted in very similar dendrograms when using various methods of analysis (Neighbor-Joining, Maximum Likelihood, Bayesian Inference), and altering the analytical parameters did not result in significant changes. Furthermore, the genetic dendrograms were strongly supported by the phenotypic and spectral analyses. These results confirm the calmodulin gene as a robust and reliable genomic marker for this group of fungi. The unsolved isolate identifications are currently under further analysis

A Cross-Sectional Population-Based Study

Funding Information: We thank the EpiDoc Unit and EpiReumaPt team for conceptualising, planning, and implementing the main research project. We would like to acknowledge that the present publication was supported by Fundação Ciência e Tecnologia, IP national support through CHRC (UIDP/04923/2020). Funding Information: This work was supported by an independent research grant from Pfizer. DC received national funding through FCT—Fundação para a Ciência e Tecnologia, I. P. under the Ph.D. grant SFRH/BD/148420/2019Background: Hip/knee osteoarthritis (HKOA) is a leading cause of disability and imposes a major socioeconomic burden. The aim of this study is to estimate the prevalence of HKOA in Portugal, characterised the clinical severity of HKOA in the population, and identified sociodemographic, lifestyle, and clinical factors associated with higher clinical and radiographic severity. Methods: Participants with a diagnosis of HKOA from the EpiReumaPt study (2011–2013) were included (n = 1,087). Hip/knee osteoarthritis diagnosis was made through a structured evaluation by rheumatologists according to American College of Rheumatology criteria. Clinical severity was classified based on Hip Disability and Osteoarthritis Outcome Scale (HOOS) and Knee Injury and Osteoarthritis Outcome Scale (KOOS) score tertiles. Radiographic severity was classified based on the Kellgren-Lawrence grades as mild, moderate, or severe. Sociodemographic lifestyle and clinical variables, including the presence of anxiety and depression symptoms, were analysed. Factors associated with higher clinical and radiographic severity were identified using ordinal logistic regression models. Results: Hip/knee osteoarthritis diagnosis was present in 14.1% of the Portuguese population [12.4% with knee osteoarthritis (OA) and 2.9% with hip OA]. Clinical severity was similar between people with hip (HOOS = 55.79 ± 20.88) and knee (KOOS = 55.33 ± 20.641) OA. People in the high HOOS/KOOS tertile tended to be older (64.39 ± 0.70 years), female (75.2%), overweight (39.0%) or obese (45.9%), and had multimorbidity (86.1%). Factors significantly associated with higher clinical severity tertile were age [55–64 years: odds ratio (OR) = 3.18; 65–74 years: OR = 3.25; ≥75 years: OR = 4.24], female sex (OR = 1.60), multimorbidity (OR = 1.75), being overweight (OR = 2.01) or obese (OR = 2.82), and having anxiety symptoms (OR = 1.83). Years of education was inversely associated with higher clinical severity. Factors significantly associated with higher radiographic severity were age (65–74 years: OR = 3.59; ≥75 years: OR = 3.05) and being in the high HOOS/KOOS tertile (OR = 4.91). Being a female and live in Lisbon or in the Centre region were inversely associated with the higher radiographic severity. Conclusion: Hip/knee osteoarthritis is present in ~1.1 million of Portuguese people. Age, educational level, and obesity are independently associated with HKOA clinical severity, whereas age, sex, geographic location, and clinical severity are independently associated with radiographic severity.publishersversionpublishe

a systematic review

Funding Information: This research received no external funding. C.S.S. is supported by the Portuguese Foundation for Science and Technology (SFRH/BD/139603/2018). The funder had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. Publisher Copyright: © 2023, BioMed Central Ltd., part of Springer Nature.Background: Translation into practice of effective physical activity interventions in primary care is difficult, due to a complex interaction of implementation determinants. We aimed to identify implementation barriers and facilitators of four primary care interventions: physical activity assessment, counselling, prescription, and referral. Methods: A systematic review of qualitative, quantitative and mixed-methods studies published since 2016 was conducted. The “Tailored Implementation for Chronic Diseases” (TICD) framework was adapted to extract and synthesize barriers and facilitators. Results: Sixty-two studies met the inclusion criteria. Barriers (n = 56) and facilitators (n = 55) were identified across seven domains, related to characteristics of the intervention, individual factors of the implementers and receivers, organizational factors, and political and social determinants. The five most frequently reported determinants were: professionals’ knowledge and skills; intervention feasibility/compatibility with primary health care routine; interventions’ cost and financial incentives; tools and materials; and professionals’ cognitions and attitudes. “Social, political and legal factors” domain was the least reported. Physical activity counselling, prescription, and referral were influenced by determinants belonging to all the seven domains. Conclusion: The implementation of physical activity interventions in primary care is influenced by a broader range of determinants. Barriers and facilitators related with health professionals, intervention characteristics, and available resources were the most frequently reported. A deep understanding of the local context, with particularly emphasis on these determinants, should be considered when preparing an intervention implementation, in order to contribute for designing tailored implementation strategies and optimize the interventions’ effectiveness.publishersversionpublishe

A experiência do bebé na creche: Perceções de mães e de educadoras no período de transição do contexto familiar para a creche

Este estudo visa contribuir para a compreensão da experiência do bebé no período de transição do ambiente familiar para a creche, analisando a perceção das mães e das educadoras acerca do estado emocional do bebé, da manutenção das rotinas e da comunicação família-creche nesse período. Mães e educadoras de 90 bebés da Grande Área Metropolitana do Porto responderam ao Questionário de Experiência na Creche na primeira e quarta semana de frequência da creche. A perceção das mães e educadoras acerca do estado emocional dos bebés, da manutenção das rotinas e comunicação família-creche foi positiva, verificando-se perceções mais positivas das educadoras relativamente ao estado emocional e à comunicação família-creche. Da primeira para a quarta semana registou-se (a) estado emocional mais positivo, percebido pelas mães e educadoras; (b) diminuição da frequência da comunicação e aumento da manutenção das rotinas, percebidas pelas mães. O estado emocional dos bebés parece estar associado à perceção da manutenção das rotinas (mães e das educadoras) e, adicionalmente, à comunicação família-creche no caso das educadoras. Este estudo parece assinalar o cuidado das famílias e educadores na transição dos bebés para a creche e destacar a importância do envolvimento das famílias e educadores para um melhor ajustamento do bebé

A resectorization of fire brigades in the north of Portugal

Sectorization can be regarded as a division of a territory into smaller regions to deal with a complex problem involving multiple-criteria. Resectorization intends to achieve another sectorization, according to some new conditions but avoiding substantial changes. An example of this can be the distribution of geographical areas by fire brigades. In Portugal, fire brigades must protect and rescue the population in the areas surrounding their fire stations. So we will use the current sectorization, the geographic and population characteristics of the areas and the fire brigades’ response capacity to provide an optimised resectorization, in order to decrease rescue time. To achieve that, we will use a decision support system using different optimisation methods, such as Non-dominated Sorting Genetic Algorithm (NSGA II), which provides an effective sectorization concerning compactness and equilibrium criteria.info:eu-repo/semantics/acceptedVersio

Creating homogeneous sectors: criteria and applications of sectorization

Sectorization is the process of grouping a set of previously defined basic units (points or small areas) into a fixed number of sectors. Sectorization is also known in the literature as districting or territory design, and is usually performed to optimize one or more criteria regarding the geographic characteristics of the territory and the planning purposes of sectors. The most common criteria are equilibrium, compactness and contiguity, which can be measured in many ways. Sectorization is similar to clustering but with a different motivation. Both aggregate smaller units into groups. But, while clustering strives for inner similarity of data, sectorization aims at outer homogeneity [1]. In clustering, groups should be very different from each other, and similar points are classified in the same cluster. In sectorization, groups should be very similar to each other, and therefore very different points can be grouped in the same sector. We classify sectorization problems into four types: basic sectorization, sectorization with service centers, resectorization, and dynamic sectorization. A Decision Support System for Sectorization, D3S, is being developed to deal with these four types of problems. Multi-objective genetic algorithms were implemented in D3S using Python, and a user-friendly web interface was developed using Django. Several applications can be solved with D3S, such as political districting, sales territory design, delivery service zones, and assignment of fire stations and health services to the population.info:eu-repo/semantics/draf