238 research outputs found

    Fluoxetine and Nutrients Removal from Aqueous Solutions by Phycoremediation

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    The tertiary treatment using microalgae offers an attractive alternative to the removal of low but relevant concentrations of pharmaceuticals from domestic wastewaters. The removal of fluoxetine from aqueous solutions by living and non-living (lyophilized) Chlorella vulgaris was assessed. The determination of the pH at the point of zero charge, Fourier transmittance infrared analysis, and scanning electron microscopy were performed to characterize the microalgae biomass. Kinetic and equilibrium experiments were performed. The pseudo-second-order model described the kinetics of fluoxetine. The corresponding kinetic constants indicated that biosorption was faster onto non-living biomass than onto living biomass. The equilibrium results showed that the systems followed the Langmuir isotherm model. The maximum capacity of living microalgae (1.9 ± 0.1 mg·g−1) was slightly higher than the non-living microalgae (1.6 ± 0.2 mg·g−1). Living Chlorella vulgaris, free and immobilized in calcium-alginate, were also used to remove fluoxetine and nutrients (nitrogen and phosphorus) from treated municipal wastewater in a batch system. In both experiments, fluoxetine was completely removed within six days. The total phosphorus (TP) and total nitrogen (TN) removal efficiencies achieved for free and immobilized cells were, null and 65.0 ± 0.1%, and 86.2 ± 0.1% and 81.8 ± 3.1, respectivelyThis research was funded by the Associate Laboratory for Green Chemistry-LAQV, which received financial support from UIDB/50006/2020, UIDP/50006/2020, and LA/P/0008/2020 by the Fundação para a Ciência e a Tecnologia (FCT)/Ministério da Ciência, Tecnologia e En sino Superior (MCTES) through national funds. This research was also funded by the EU and FCT/UEFISCDI/FORMAS, in the frame of the collaborative international consortium REWATER— “Sustainable and safe water management in agriculture: increasing the efficiency of water reuse for crop growth while protecting ecosystems, services and citizens’ welfare” (WaterJPI/0007/2016), which was financed under the ERA-NET Co-fund WaterWorks2015 Call, as an integral part of the 2016 Joint Activities developed by the Water Challenges for a Changing World Joint Program Initiative (Water JPI). The research was funded also by FCT and BiodivRestore Joint Call 2020–2021-European Union’s Horizon 2020 research and innovation program under grant agreement No. 101003777- BiodivRestore-406/DivRestore/0002/2020-BioReset-“Biodiversity restoration and conservation of inland water ecosystems for environmental and human well-being”. A.D.M. Silva would like to thank FCT for her Ph.D. Grant SFRH/BD/138/780/2018. The authors are greatly indebted to all financing sources. The authors are grateful to Materials Centre of the University of Porto (CEMUP), Porto, Portugal, for expert assistance with SEM/EDSinfo:eu-repo/semantics/publishedVersio

    Screening of perfused combinatorial 3D microenvironments for cell culture

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    Biomaterials combining biochemical and biophysical cues to establish close-to-extracellular matrix (ECM) models have been explored for cell expansion and differentiation purposes. Multivariate arrays are used as material-saving and rapid-to-analyze platforms, which enable selecting hit-spotted formulations targeting specific cellular responses. However, these systems often lack the ability to emulate dynamic mechanical aspects that occur in specific biological milieus and affect physiological phenomena including stem cells differentiation, tumor progression, or matrix modulation. We report a tailor-made strategy to address the combined effect of flow and biochemical composition of three-dimensional (3D) biomaterials on cellular response. We suggest a simple-to-implement device comprising (i) a perforated platform accommodating miniaturized 3D biomaterials and (ii) a bioreactor that enables the incorporation of the biomaterial-containing array into a disposable perfusion chamber. The system was upscaled to parallelizable setups, increasing the number of analyzed platforms per independent experiment. As a proof-of-concept, porous chitosan scaffolds with 1 mm diameter were functionalized with combinations of 5 ECM and cell-cell contact-mediating proteins, relevant for bone and dental regeneration, corresponding to 32 protein combinatorial formulations. Mesenchymal stem cells adhesion and production of an early osteogenic marker were assessed on-chip on static and under-flow dynamic perfusion conditions. Different hit-spotted biomaterial formulations were detected for the different flow regimes using direct image analysis. Cell-binding proteins still poorly explored as biomaterials components amelogenin and E-cadherin - were here shown as relevant cell response modulators. Their combination with ECM cell-binding proteins - fibronectin, vitronectin, and type 1 collagen - rendered specific biomaterial combinations with high cell adhesion and ALP production under flow. The developed versatile system may be targeted at wM.B. Oliveira acknowledges the financial support from Portuguese Foundation for Science and Technology- FCT (Grant SFRH/BPD/111354/2015). This work was developed within the scope of the projects CICECO-Aveiro Institute of Materials, POCI-01-0145-FEDER-007679 (FCT Ref. UID/CTM/50011/2013) and IPC/i3N Minho (FCT Ref. UID/CTM/50025/2013), financed by national funds through the FCT/MEC and when appropriate co-financed by FEDER under the PT2020 Partnership Agreement. This work was also supported by European Research Council grant agreement ERC-2014-ADG-669858 (project ATLAS)

    Bologna process and its implications: what does the research literature tell us?

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    "Apresentação efetuada na 55ª Assembleia Mundial da International Council on Education for Teaching (ICET), em Glasgow, Scotland, 2011

    The amino acids motif-32GSSYN36-in the catalytic domain of E. coli flavorubredoxin NO reductase is essential for its activity

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    Funding Information: Funding: This study was financially supported by the Portuguese Fundação para a Ciência e Tec-nologia (FCT), grants PTDC/BIA-BQM/27959/2017 and PTDC/BIA-BQM/0562/2020, and Project MOSTMICRO-ITQB with references UIDB/04612/2020 and UIDP/04612/2020. This project has also received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement 810856. MCM is the recipient of FCT grant SFRH/BD/143651/2019. BAS is the recipient of FCT grant DFA/BD/8066/2020. Funding Information: This study was financially supported by the Portuguese Funda??o para a Ci?ncia e Tecnologia (FCT), grants PTDC/BIA-BQM/27959/2017 and PTDC/BIA-BQM/0562/2020, and Project MOSTMICRO-ITQB with references UIDB/04612/2020 and UIDP/04612/2020. This project has also received funding from the European Union?s Horizon 2020 research and innovation program under grant agreement 810856. MCM is the recipient of FCT grant SFRH/BD/143651/2019. BAS is the recipient of FCT grant DFA/BD/8066/2020. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Flavodiiron proteins (FDPs) are a family of modular and soluble enzymes endowed with nitric oxide and/or oxygen reductase activities, producing N2O or H2O, respectively. The FDP from Escherichia coli, which, apart from the two core domains, possesses a rubredoxin-like domain at the C-terminus (therefore named flavorubredoxin (FlRd)), is a bona fide NO reductase, exhibiting O2 reducing activity that is approximately ten times lower than that for NO. Among the flavorubredoxins, there is a strictly conserved amino acids motif,-G[S,T]SYN-, close to the catalytic diiron center. To assess its role in FlRd’s activity, we designed several site-directed mutants, replacing the conserved residues with hydrophobic or anionic ones. The mutants, which maintained the general characteristics of the wild type enzyme, including cofactor content and integrity of the diiron center, revealed a decrease of their oxygen reductase activity, while the NO reductase activity—specifically, its physiological function—was almost completely abolished in some of the mutants. Molecular modeling of the mutant proteins pointed to subtle changes in the predicted structures that resulted in the reduction of the hydration of the regions around the conserved residues, as well as in the elimination of hydrogen bonds, which may affect proton transfer and/or product release.publishe

    Quinoline- and benzoselenazole-derived unsymmetrical squaraine cyanine dyes: design, synthesis, photophysicochemical features and light-triggerable antiproliferative effects against breast cancer cell lines

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    Photodynamic therapy is an innovative treatment approach broadly directed towards oncological diseases. Its applicability and efficiency are closely related to the interaction of three main components, namely a photosensitizer, light and molecular triplet oxygen, which should drive cell death. Recently, several studies have demonstrated that squaraine cyanine dyes have a set of photophysical and photochemical properties that have made of these compounds’ potential photosensitizers for this therapeutic modality. In the present research work, we describe the synthesis and characterization of four quinoline- and benzoselenazole-derived unsymmetrical squaraine cyanine dyes. Except for the precursor of aminosquaraine dyes, i.e., O-methylated derivative, all dyes were evaluated for their behavior and absorption capacity in different organic and aqueous solvents, their ability to form singlet oxygen, their light-stability, and in vitro phototherapeutic effects against two human breast cancer cell cultures (BT-474 and MCF-7). Regardless of the nature of the used solvents, the synthesized dyes showed intense absorption in the red and near-infrared spectral regions, despite the formation of aggregates in aqueous media. Dyes showed high light-stability against light exposure. Despite the low ability to produce singlet oxygen, aminosquaraine dyes demonstrated worthy in vitro phototherapeutic activity.This research was funded by the European Investment Funds by FEDER/COMPETE/POCI under projects POCI-01-0145-FEDER-006958 (CITAB) and POCI-01-0145-FEDER-007491 (CICS-UBI) and Funds by FCT—Portuguese Foundation for Science and technology, under the projects UIDB/ 04033/2020 (CITAB) and UIDB/ 00616/2020 (CQ-VR). This work was also supported by funds from the Health Sciences Research Center (CICS-UBI) through National Funds by FCT—Foundation for Science and Technology (UID/Multi/00709/2019).The research at iBB was supported by Project UID/NAN/50024/2019 and M-ERA-NET/0002/2015 from FCT. E.L. was supported by the FCT PhD grant SFRH/BD/147645/2019.info:eu-repo/semantics/publishedVersio

    Is the chlorophyll derivative Zn(II)e6Me a good photosensitizer to be used in root canal disinfection?

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    The aim of this study was to assess antimicrobial efficacy and cytotoxic outcomes of a chlorophyll based photosensitizer (PS) Zn(II)chlorin e6 methyl ester (Zn(II)e6Me), when applied to human dentin discs and root blocks infected with 48 h biofilms. The results were compared with the ones obtained with FotoSan® (commercial Toluidine Blue O formulation) and 3% sodium hypochlorite (NaOCl).publishe

    Red and near-infrared absorbing dicyanomethylene squaraine cyanine dyes: photophysicochemical properties and anti-tumor photosensitizing effects

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    Photodynamic therapy is a medical modality developed for the treatment of several diseases of oncological and non-oncological etiology that requires the presence of a photosensitizer, light and molecular oxygen, which combined will trigger physicochemical reactions responsible for reactive oxygen species production. Given the scarcity of photosensitizers that exhibit desirable characteristics for its potential application in this therapeutic strategy, the main aims of this work were the study of the photophysical and photochemical properties and the photobiological activity of several dicyanomethylene squaraine cyanine dyes. Thus, herein, the study of their aggregation character, photobleaching and singlet oxygen production ability, and the further application of the previously synthesized dyes in Caco-2 and HepG2 cancer cell lines, to evaluate their phototherapeutic effects, are described. Dicyanomethylene squaraine dyes exhibited moderate light-stability and, despite the low singlet oxygen quantum yields, were a core of dyes that exhibited relevant in vitro photodynamic activity, as there was an evident increase in the toxicity of some of the tested dyes exclusive to radiation treatments.This research was funded by the European Investment Funds by FEDER/COMPETE/POCI under projects POCI-01-0145-FEDER-006958 (CITAB) and POCI-01-0145-FEDER-007491 (CICS-UBI) and Funds by FCT – Portuguese Foundation for Science and technology, under the projects UIDB/04033/2019 (CITAB) and UIDB/00616/2020 (CQ-VR). This work was also supported by funds from the Health Sciences Research Center (CICS-UBI) through National Funds by FCT—Foundation for Science and Technology (UID/Multi/00709/2019). The research at CQFM was supported by Project UID/NAN/50024/2019 and M-ERA-NET/0002/2015 from FCT. E. L. was supported by the FCT PhD grant SFRH/BD/147645/2019

    Concern-Driven Integrated Toxicity Testing Strategies for Nanomaterials - Report of the NanoSafety Cluster Working Group 10

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    Bringing together topic-related European Union-(EU)-funded projects, the so-called “NanoSafety Cluster” aims at identifying key areas for further research on risk assessment procedures for nanomaterials (NM). The outcome of NanoSafety Cluster Working Group 10, this commentary presents a vision for concern-driven integrated approaches for the (eco-)toxicological testing and assessment (IATA) of NM. Such approaches should start out by determining concerns, i.e. specific information needs for a given NM based on realistic exposure scenarios. Recognized concerns can be addressed in a set of tiers using standardized protocols for NM preparation and testing. Tier 1 includes determining physico-chemical properties, non-testing (e.g. structure activity relationships) and evaluating existing data. In tier 2, a limited set of in vitro and in vivo tests are performed that can either indicate that the risk of the specific concern is sufficiently known or indicate the need for further testing, including details for such testing. Ecotoxicological testing begins with representative test organisms followed by complex test systems. After each tier, it is evaluated whether the information gained permits assessing the safety of the NM so that further testing can be waived. By effectively exploiting all available information, IATA allow accelerating the risk assessment process and reducing testing costs and animal use (in line with the 3Rs principle implemented in EU Directive 2010/63/EU). Combining material properties, exposure, biokinetics, and hazard data, information gained with IATA can be used to recognize groups of NM based upon similar modes-of-action. Grouping of substances in return should form integral part of the IATA themselves

    Severe COVID-19 recovery is associated with timely acquisition of a myeloid cell immune-regulatory phenotype

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    Copyright © 2021 Trombetta, Farias, Gomes, Godinho-Santos, Rosmaninho, Conceição, Laia, Santos, Almeida, Mota, Gomes, Serrano, Veldhoen, Sousa and Fernandes. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.After more than one year since the COVID-19 outbreak, patients with severe disease still constitute the bottleneck of the pandemic management. Aberrant inflammatory responses, ranging from cytokine storm to immune-suppression, were described in COVID-19 and no treatment was demonstrated to change the prognosis significantly. Therefore, there is an urgent need for understanding the underlying pathogenic mechanisms to guide therapeutic interventions. This study was designed to assess myeloid cell activation and phenotype leading to recovery in patients surviving severe COVID-19. We evaluated longitudinally patients with COVID-19 related respiratory insufficiency, stratified according to the need of intensive care unit admission (ICU, n = 11, and No-ICU, n = 9), and age and sex matched healthy controls (HCs, n = 11), by flow cytometry and a wide array of serum inflammatory/immune-regulatory mediators. All patients featured systemic immune-regulatory myeloid cell phenotype as assessed by both unsupervised and supervised analysis of circulating monocyte and dendritic cell subsets. Specifically, we observed a reduction of CD14lowCD16+ monocytes, and reduced expression of CD80, CD86, and Slan. Moreover, mDCs, pDCs, and basophils were significantly reduced, in comparison to healthy subjects. Contemporaneously, both monocytes and DCs showed increased expression of CD163, CD204, CD206, and PD-L1 immune-regulatory markers. The expansion of M2-like monocytes was significantly higher at admission in patients featuring detectable SARS-CoV-2 plasma viral load and it was positively correlated with the levels of specific antibodies. In No-ICU patients, we observed a peak of the alterations at admission and a progressive regression to a phenotype similar to HCs at discharge. Interestingly, in ICU patients, the expression of immuno-suppressive markers progressively increased until discharge. Notably, an increase of M2-like HLA-DRhighPD-L1+ cells in CD14++CD16- monocytes and in dendritic cell subsets was observed at ICU discharge. Furthermore, IFN-γ and IL-12p40 showed a decline over time in ICU patients, while high values of IL1RA and IL-10 were maintained. In conclusion, these results support that timely acquisition of a myeloid cell immune-regulatory phenotype might contribute to recovery in severe systemic SARS-CoV-2 infection and suggest that therapeutic agents favoring an innate immune system regulatory shift may represent the best strategy to be implemented at this stage.The Research was funded by Fundação para a Ciência e Tecnologia (FCT), “APOIO ESPECIAL RESEARCH 4COVID-19” projects 803, 125, 231_596873172, and 729. AMCG and GF received fellowships funded by FCT (DOCTORATES4COVID-19, 2020.10202.BD), and JANSSEN- CILAG FARMACÊUTICA, respectively. The funder was not involved in the study design, collection, analysis, interpretation of data, writing of the article or decision to submit it for publication. MV was supported by the European Union H2020 ERA project (No 667824 – EXCELLtoINNOV). This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 667824.info:eu-repo/semantics/publishedVersio
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