Regulatory crosstalk by protein kinases on CFTR trafficking and activity

Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a member of the ATP binding cassette (ABC) transporter superfamily that functions as a cAMP-activated chloride ion channel in fluid-transporting epithelia. There is abundant evidence that CFTR activity (i.e., channel opening and closing) is regulated by protein kinases and phosphatases via phosphorylation and dephosphorylation. Here, we review recent evidence for the role of protein kinases in regulation of CFTR delivery to and retention in the plasma membrane. We review this information in a broader context of regulation of other transporters by protein kinases because the overall functional output of transporters involves the integrated control of both their number at the plasma membrane and their specific activity. While many details of the regulation of intracellular distribution of CFTR and other transporters remain to be elucidated, we hope that this review will motivate research providing new insights into how protein kinases control membrane transport to impact health and disease.Work supported by the Portuguese Fundação para a Ciência e Tecnologia (fellowship grant SFRH/BSAB/105741/2014—to CF, grant PTDC/SAU-ORG/119782/2010—to PJ, and centre grant UID/MULTI/04046/2013—to BioISI); Gilead Génese PGG/039/2014 and ERS Romain Pauwels 2012 (to CF); CFF SWIATE14P0 (to AS); NIH NIDDK P30 072506 Basic and Translational Studies of Cystic Fibrosis (P&F to AS)

Cell type-specific regulation of CFTR trafficking—on the verge of progress

Trafficking of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein is a complex process that starts with its biosynthesis and folding in the endoplasmic reticulum. Exit from the endoplasmic reticulum (ER) is coupled with the acquisition of a compact structure that can be processed and traffic through the secretory pathway. Once reaching its final destination—the plasma membrane, CFTR stability is regulated through interaction with multiple protein partners that are involved in its post-translation modification, connecting the channel to several signaling pathways. The complexity of the process is further boosted when analyzed in the context of the airway epithelium. Recent advances have characterized in detail the different cell types that compose the surface epithelium and shifted the paradigm on which cells express CFTR and on their individual and combined contribution to the total expression (and function) of this chloride/bicarbonate channel. Here we review CFTR trafficking and its relationship with the knowledge on the different cell types of the airway epithelia. We explore the crosstalk between these two areas and discuss what is still to be clarified and how this can be used to develop more targeted therapies for CF

Impact of different aquatic exercise programs on body composition, functional fitness and cognitive function of non-institutionalized elderly adults: a randomized controlled trial

Aquatic physical exercise programs have become progressively more popular among elderly people. Some of the major physical exercise program disadvantages on land are minimized due to the specific properties of the aquatic environment. The purpose of the present randomized controlled study is to verify the effects of different aquatic physical exercise programs on body composition, functional fitness and cognitive function in non-institutionalized elderly people. For this study, 102 elderly individuals were randomly allocated into four different groups: AerG (n = 25, 71.44 ± 4.84 years); IntG (n = 28, 72.64 ± 5.22 years); ComG (n = 29, 71.90 ± 5.67 years) and CG (n = 20, 73.60 ± 5.25 years). Individuals from the groups AerG, IntG and ComG participated in three different aquatic physical exercise programs for a period of 28 weeks. The CG participants kept to their usual routines. All participants were evaluated for body composition, functional fitness and cognitive function at two time moments, i.e., pre- (M1) and post-intervention (M2). Significant differences for body composition were found between M1 and M2 for FM (p < 0.001), LBM (p < 0.001) and WCir (p < 0.01) in the AerG, for BMI (p < 0.05), FM (p < 0.05), LBM (p < 0.001) and LCir-R (p < 0.05) in the IntG, and for WGT (p < 0.01), FM (p < 0.05), LBM (p < 0.01), LCir-R (p < 0.05) and LCir-L (p < 0.01) in the ComG groups. For functional fitness, differences were found between M1 and M2 for 2m-ST (p < 0.000), 30s-CS (p < 0.000), 30s-AC (p < 0.05), HG-T-R (p < 0.000) and HG-T-L (p < 0.000) in the AerG, for 2m-ST (p < 0.05), BS-R (p < 0.05), 30s-CS (p < 0.000), 30s-AC(p < 0.01), HG-T-R (p < 0.000) and HG-T-L (p < 0.000) in the IntG, and for 30s-CS (p < 0.000), HG-T-R (p < 0.000) and HG-T-L (p < 0.000) in the ComG groups. The present study evidenced the beneficial effects of physical exercise in an aquatic environment on body composition, functional fitness and cognitive function in non-institutionalized elderly adults. The ComG water-based exercise program showed more beneficial effects in the improvement of body composition and cognitive function variables, while the IntG and AerG programs were more effective in the improvement of functional fitness.info:eu-repo/semantics/publishedVersio

Network Biology Identifies Novel Regulators of CFTR Trafficking and Membrane Stability

Free PMC article: https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/31231217/In cystic fibrosis, the most common disease-causing mutation is F508del, which causes not only intracellular retention and degradation of CFTR, but also defective channel gating and decreased membrane stability of the small amount that reaches the plasma membrane (PM). Thus, pharmacological correction of mutant CFTR requires targeting of multiple cellular defects in order to achieve clinical benefit. Although small-molecule compounds have been identified and commercialized that can correct its folding or gating, an efficient retention of F508del CFTR at the PM has not yet been explored pharmacologically despite being recognized as a crucial factor for improving functional rescue of chloride transport. In ongoing efforts to determine the CFTR interactome at the PM, we used three complementary approaches: targeting proteins binding to tyrosine-phosphorylated CFTR, protein complexes involved in cAMP-mediated CFTR stabilization at the PM, and proteins selectively interacting at the PM with rescued F508del-CFTR but not wt-CFTR. Using co-immunoprecipitation or peptide-pull down strategies, we identified around 400 candidate proteins through sequencing of complex protein mixtures using the nano-LC Triple TOF MS technique. Key candidate proteins were validated for their robust interaction with CFTR-containing protein complexes and for their ability to modulate the amount of CFTR expressed at the cell surface of bronchial epithelial cells. Here, we describe how we explored the abovementioned experimental datasets to build a protein interaction network with the aim of identifying novel pharmacological targets to rescue CFTR function in cystic fibrosis (CF) patients. We identified and validated novel candidate proteins that were essential components of the network but not detected in previous proteomic analyses.This work was supported by FCT, Portugal, through center grant UID/MULTI/04046/2019 to BioISI and the BioSys PhD program PD65-2012 (fellowships SFRH/BD/52488/2014, SFRH/ BD/106084/2015, and SFRH/BD/52490/2014 to CL, JS, and AM, respectively).info:eu-repo/semantics/publishedVersio

Reversible imine crosslinking in waterborne self-healing polymer coatings

Waterborne polymer coatings have the potential to address the environmental concerns associated with solvent based systems. To improve their performance without using volatile organic compounds, we propose a new approach based on reconfigurable covalent crosslinking that provides mechanical resistance and self-healing properties. The new waterborne polymer coatings are based on mixtures of aldehyde- and amine-functionalized polymer nanoparticles (PNPs) that take advantage of the reversibility of imine bonds in the presence of water. Different degrees of functional monomer incorporation (10 % to 40 %) allowed us to balance crosslinking and interdiffusion during film formation, to obtain mechanically robust and solvent resistant films. A clear structure-properties relation was assessed by following the formation of water resulting from amine-aldehyde condensation crosslinking, measured by differential scanning calorimetry. The resulting polymer coatings further show self-healing properties at room temperature, triggered with residual amounts of water and featuring high recovery rates of the mechanical properties. Our mechanically robust waterborne polymer coatings based in imine reversible crosslinking, featuring self-healing in mild conditions, offer excellent prospects for application in smart coating materials.publishe

Inequality and crisis: conspicuous consumption as the missing link in the portuguese case

Portuguese household debt increased above GDP between 2000 and 2007. This article uses conspicuous consumption to explain credit demand dynamics. The author develops an institutionalist framework and consider how rapid high inequalities and increasing top income share favored conspicuous consumption and climbing household debt.info:eu-repo/semantics/publishedVersio

Multifunctional platform based on electroactive polymers and silica nanoparticles for tissue engineering applications

Poly(vinylidene fluoride) nanocomposites processed with different morphologies, such as porous and non-porous films and fibres, have been prepared with silica nanoparticles (SiNPs) of varying diameter (17, 100, 160 and 300 nm), which in turn have encapsulated perylenediimide (PDI), a fluorescent molecule. The structural, morphological, optical, thermal, and mechanical properties of the nanocomposites, with SiNP filler concentration up to 16 wt %, were evaluated. Furthermore, cytotoxicity and cell proliferation studies were performed. All SiNPs are negatively charged independently of the pH and more stable from pH 5 upwards. The introduction of SiNPs within the polymer matrix increases the contact angle independently of the nanoparticle diameter. Moreover, the smallest ones (17 nm) also improve the PVDF Youngs modulus. The filler diameter, physico-chemical, thermal and mechanical properties of the polymer matrix were not significantly affected. Finally, the SiNPs inclusion does not induce cytotoxicity in murine myoblasts (C2C12) after 72 h of contact and proliferation studies reveal that the prepared composites represent a suitable platform for tissue engineering applications, as they allow us to combine the biocompatibility and piezoelectricity of the polymer with the possible functionalization and drug encapsulation and release of the SiNP.This work was supported by the Portuguese Foundation for Science and Technology (FCT) in the framework of the Strategic Funding UID/FIS/04650/2013 and UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) and project POCI-01-0145-FEDER-028237 funded by national funds through Fundação para a Ciência e a Tecnologia (FCT) and by the ERDF through the COMPETE2020-Programa Operacional Competitividade e Internacionalização (POCI); and also under the scope of the strategic funding of UID/BIO/04469 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020-Programa Operacional Regional do Norte. The authors also thank the FCT for the SFRH/BD/111478/2015 (S.R.), SFRH/BPD/96707/2013 (T.R.), SFRH/BPD/90870/2012 (C.R.) and SFRH/BPD/121526/2016 (D.C) grants. The authors acknowledge funding from the Spanish Ministry of Economy and Competitiveness (MINECO) through the project MAT2016-76039-C4-3-R (AEI/FEDER, UE) and from the Basque Government Industry and Education Departments under the ELKARTEK, HAZITEK and PIBA (PIBA-2018-06) programs, respectively.info:eu-repo/semantics/publishedVersio

Adenine interaction with and adsorption on Fe-ZSM-5 zeolites: A prebiotic chemistry study using different techniques

Most adsorption experiments are performed under conditions that did not exist on Earth before the life arose on it. Because adsorption is the first step for all other processes (protection against degradation and polymerization), it is important that it is performed under conditions that existed on prebiotic Earth. In this paper, we use an artificial seawater (seawater 4.0 Ga), which contains major cations and anions that could present on the oceans of the prebiotic Earth. In addition, zeolites, with substituted Fe in the framework, and adenine were probably common substances on the prebiotic Earth. Thus, study the interaction between them is an important issue in prebiotic chemistry. There are two main findings described in this paper. Firstly, zeolites with different Si/Fe ratios adsorbed adenine differently. Secondly, XAFS showed that, after treatments with seawater 4.0 Ga and adenine, an increase in the complexity of the system occurred. In general, salts of seawater 4.0 Ga did not affect the adsorption of adenine onto zeolites and adenine adsorbed less onto zeolites with iron isomorphically substituted. The C=C and NH2 groups of adenine interacted with the zeolites. Gypsum, formed from aqueous species dissolved in seawater 4.0 Ga, precipitated onto zeolites. EPR spectra of zeolites showed lines caused by Fe framework and Fe3+ species. TG curves of zeolites showed events caused by loss of water weakly bound to zeolite (in the 30-140 °C range), water bounded to iron species or cations from seawater 4.0 Ga or located in the cavities of zeolites (157-268 °C) and degradation of adenine adsorbed onto zeolites (360-600 °C). Mass loss follows almost the same order as the amount of adenine adsorbed onto zeolites. The XAFS spectrum showed that Fe3+ could be substituted into the framework of the Fe7-ZSM-5 zeolite

description of the methodological approach

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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