Browsing by Author "Costa, Ana M. Rosa da"
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- A bridge between liquid-liquid extraction and the use of bacterial communities for palladium and platinum recovery as nanosized metal sulphidesPublication . Assunção, Ana; Matos, Ana; Costa, Ana M. Rosa da; Candeias, Antonio; Costa, Maria ClaraThe Platinum Group Metals (PGM) are rare in the Earth's crust and in the past years their use had a considerable expansion limiting their availability, which justifies PGM's high commercial value and demand. Therefore, PGM recovery from secondary sources is very important from both economic and environmental points of view. In recent years, several methods for PGM removal have been investigated. Our research group has been studying the removal/recovery of PGM using both chemical (liquid liquid extraction) and biological methods (using anaerobic bacterial communities).The aim of this study was to combine these two chemical and biological approach technologies, for PGM recovery. For that purpose, Pd(II) and Pt(IV) present in aqueous phases, were extracted to organic phases composed by N,N'-dimethyl-N,N'-dicyclohexylsuccinamide (DMDCHSA) and N,N'-dimethyl-N,N'-dicyclohexyltetradecylmalonamide (DMDCHTDMA) in 1,2-dichloroethane, respectively, with an extraction efficiency of 79% for Pd(II) and 99% for Pt(IV). The metals in the loaded organic phases were then stripped with seawater, a low-cost solution largely available in nature, with efficiencies of 100% for Pd and 86% for Pt. Finally, the metals were precipitated and recovered using metabolic products produced by a community enriched for sulphate-reducing bacteria. The palladium precipitated completely, while 86% of platinum was precipitated. During the precipitation process sulphide concentration in the solution decreased and the analysis of the precipitates was consistent with the formation of nanosized PdS and PtS2.To our knowledge, this research shows, for the first time, the potential of combining liquid liquid extraction with the use of bacteria aiming platinum and palladium recovery, as metal sulphides, from aqueous media. (C) 2016 Elsevier B.V. All rights reserved.
- Antioxidant and anti-cholinesterase activities of Lavandula viridis L’Hér extracts after in vitro gastrointestinal digestionPublication . Costa, Patrícia; Grevenstuk, Tomás; Costa, Ana M. Rosa da; Gonçalves, Sandra; Romano, AnabelaLavandula viridis L’Hér is an aromatic shrub with relevant biological activities associated to the presence ofphenolic compounds. However, these compounds must be bioavailable to exert their biological function.Therefore, in this study, we investigated the influence of the L. viridis extract matrices on the digestion ofthe main component rosmarinic acid and their antioxidant and anti-cholinesterase activities after in vitrogastrointestinal digestion. Overall, the antioxidant and anti-cholinesterase activities were assured afterin vitro gastrointestinal processes and we observed that the L. viridis extract matrices have an importantrole in the bioactive effects of their main compound, rosmarinic acid. The L. viridis extracts and rosmarinicacid did not show any toxic effect on colon adenocarcinoma Caco-2 cell viability. Finally, the compoundsfrom L. viridis extracts were not metabolized by Caco-2 cells and were not able to permeate into them.
- Biocompatibility and stability of polysaccharide polyelectrolyte complexes aimed at respiratory deliveryPublication . Rodrigues, Susana; Cardoso, Lurdes; Costa, Ana M. Rosa da; Grenha, AnaChitosan (CS) and chondroitin sulfate (CHS) are natural polymers with demonstrated applicability in drug delivery, while nanoparticles are one of the most explored carriers for transmucosal delivery of biopharmaceuticals. In this work we have prepared CS/CHS nanoparticles and associated for the first time the therapeutic protein insulin. Fluorescein isothiocyanate bovine serum albumin (FITC-BSA) was also used to enable comparison of behaviors regarding differences in molecular weight (5.7 kDa versus 67 kDa). Nanoparticles of approximately 200 nm and positive zeta potential around +20 mV were obtained. These parameters remained stable for up to 1 month at 4 C. Proteins were associated with efficiencies of more than 50%. The release of FITC-BSA in PBS pH 7.4 was more sustained (50% in 24 h) than that of insulin (85% in 24 h). The biocompatibility of nanoparticles was tested in Calu-3 and A549 cells by means of three different assays. The metabolic assay MTT, the determination of lactate dehydrogenase release, and the quantification of the inflammatory response generated by cell exposure to nanoparticles have indicated an absence of overt toxicity. Overall, the results suggest good indications on the application of CS/CHS nanoparticles in respiratory transmucosal protein delivery, but the set of assays should be widened to clarify obtained results.
- Characterization and comparison of two novel nanosystems associated with siRNA for cellular therapyPublication . André, E. M.; Pensado, A.; Resnier, P.; Braz, L.; Costa, Ana M. Rosa da; Passirani, C.; Sanchez, A.; Montero-Menei, C. N.To direct stem cell fate, a delicate control of gene expression through small interference RNA (siRNA) is emerging as a new and safe promising strategy. In this way, the expression of proteins hindering neuronal commitment may be transiently inhibited thus driving differentiation. Mesenchymal stem cells (MSC), which secrete tissue repair factors, possess immunomodulatory properties and may differentiate towards the neuronal lineage, are a promising cell source for cell therapy studies in the central nervous system. To better drive their neuronal commitment the repressor Element-1 silencing transcription (REST) factor, may be inhibited by siRNA technology. The design of novel nanoparticles (NP) capable of safely delivering nucleic acids is crucial in order to successfully develop this strategy. In this study we developed and characterized two different siRNA NP. On one hand, sorbitan monooleate (Span(®)80) based NP incorporating the cationic components poly-l-arginine or cationized pullulan, thus allowing the association of siRNA were designed. These NP presented a small size (205nm) and a negative surface charge (-38mV). On the other hand, lipid nanocapsules (LNC) associating polymers with lipids and allowing encapsulation of siRNA complexed with lipoplexes were also developed. Their size was of 82nm with a positive surface charge of +7mV. Both NP could be frozen with appropriate cryoprotectors. Cytotoxicity and transfection efficiency at different siRNA doses were monitored by evaluating REST expression. An inhibition of around 60% of REST expression was observed with both NP when associating 250ng/mL of siRNA-REST, as recommended for commercial reagents. Span NP were less toxic for human MSCs than LNCs, but although both NP showed a similar inhibition of REST over time and the induction of neuronal commitment, LNC-siREST induced a higher expression of neuronal markers. Therefore, two different tailored siRNA NP offering great potential for human stem cell differentiation have been developed, encouraging the pursuit of further in vitro and in vivo in studies.
- Charged pullulan derivatives for the development of nanocarriers by polyelectrolyte complexationPublication . Dionísio, Marita; Braz, L.; Corvo, M.; Lourenço, J. P.; Grenha, Ana; Costa, Ana M. Rosa daPullulan, a neutral polysaccharide, was chemically modified in order to obtain two charged derivatives: reaction with SO3(.)DMF complex afforded a sulfate derivative (SP), while reaction with glycidyltrimethylammonium chloride gave a quaternary ammonium salt (AP). The presence of the charged groups was confirmed by FTIR. Assessment of the positions where the reaction took place was based on (1)H- and (13)C NMR (COSY, HSQC-TOCSY, HSQC-DEPT, and HMBC) experiments. Estimation of the degree of substitution (DS) was made from elemental analysis data, and further confirmed by NMR peak areas in the case of AP. These new derivatives showed the capability to condense with each other, forming nanoparticles with the ability to associate a model protein (BSA) and displaying adequate size for drug delivery applications, therefore making them good candidates for the production of pullulan-based nanocarriers by polyelectrolyte complexation.
- Chitosan/carrageenan nanoparticles: effect of cross-linking with tripolyphosphatePublication . Rodrigues, Susana; Costa, Ana M. Rosa da; Grenha, AnaChitosan/carrageenan/tripolyphosphate nanoparticles were prepared by polyelectrolyte complexation/ionic gelation, the latter compound acting as cross-linker. The incorporation of the three components in the nanoparticle matrix was assessed by analytical techniques (FTIR, XPS and TOF-SIMS). Using chitosan/carrageenan nanoparticles as control, the effect of the cross-linker in the particles properties was studied. A decrease in size (from 450-500 nm to 150-300 nm) and in zeta potential (from +75 - +85 mV to +50 - +60 mV), and an increase in production yield (from 15-20% to 25-35%), and in stability (from one week to up to 9 months) were observed. Also, a correlation between positive to negative charge ratios in the formulations and the above characteristics was established. The small size and high positive surface charge make the developed chitosan/carrageenan/tripolyphosphate nanoparticles potential tools for an application in mucosal delivery of macromolecules.
- Determination of the botanical origin of honey by sensor fusion of impedance e-tongue and optical spectroscopyPublication . Ulloa, P. A.; Guerra, Rui Manuel Farinha das Neves; Cavaco, A. M.; Costa, Ana M. Rosa da; Figueira, A.C.; Fernandes, A.The aim of this study was to discriminate four commercial brands of Portuguese honeys according to their botanical origin by sensor fusion of impedance electronic tongue (e-tongue) and optical spectroscopy (UV–Vis–NIR) assisted by Principal Component Analysis (PCA) and Cluster Analysis (CA). We have also introduced a new technique for variable selection through one-dimensional clustering which proved very useful for data fusion. The results were referenced against standard sample identification by classical melissopalynology analysis. Individual analysis of each technique showed that the e-tongue clearly outperformed the optical techniques. The electronic and optical spectra were fitted to analytical models and the model coefficients were used as new variables for PCA and CA. This approach has improved honey classification by the e-tongue but not by the optical methods. Data from the three techniques was then considered simultaneously. Simple concatenation of all matrices did not improve the classification results. Multi-way PCA (MPCA) proved to be a good option for data fusion yielding 100% classification success. Finally, a variable selection method based on one-dimensional clustering was used to define two new approaches to sensor fusion, and both yielded sample clusters even better defined than using MPCA. In this work we demonstrate for the first time the feasibility of sensor fusion of electronic and optical spectroscopy data and propose a new variable selection method that improved significantly the classification of the samples through multivariate statistical analysis.
- Effect of monomer organization on the behavior of PSS-PS micellesPublication . Alves, Nuno G.; Rocha, Luciana S.; Moreira, José; Costa, Ana M. Rosa da; Pinheiro, J. P.Six copolymers of styrenesulfonate-styrene (SS-S) of varying molecular weights were prepared by reversible addition-fragmentation chain transfer (RAFT) polymerization and proven to give rise to micelles in aqueous solution. Three are diblock copolymers (PSS-b-PS), while the other three have a random distribution of the two monomers, probably forming a gradient (PSS-co-PS). The latter ones form micelles of smaller sizes and present lower critical micellar concentrations (CMC) than the former. The morphologic and dimensional features of the micelles were obtained by TEM analysis and their CMC were determined by fluorescence spectroscopy.The trace metal ion removal ability of these micelles was tested. Thus the complexation capacity of PSS-b-PS and PSS-co-PS micelles towards Cd and Pb was evaluated using an electro-analytical stripping technique (AGNES). Both micelles performed similarly yielding percentages of metal removal that varied between 85 and 97% and 95-99%, for Cd(II) and Pb(II) respectively. Both copolymers demonstrate a good potential for future application in metal removal from water, namely on Micellar Enhanced Ultrafiltration techniques. (C) 2016 Elsevier B.V. All rights reserved.
- Efficiency of RAFT-synthesized PDMAEMA in gene transfer to the retinaPublication . Bitoque, Diogo; S, Simão; Oliveira, Ana V.; Machado, S.; Duran, Margarita R.; Lopes, Eduardo; Costa, Ana M. Rosa da; Silva, GabrielaGene therapy has long been heralded as the new hope to evolve from symptomatic care of genetic pathologies to a full cure. Recent successes in using gene therapy for treating several ocular and haematopoietic pathologies have shown the great potential of this approach that, in the early days, relied on the use of viral vectors, which were considered by many to be undesirable for human treatment. Therefore, there is considerable interest and effort in developing non-viral vectors, with efficiency close to that of viral vectors. The aim of this study was to develop suitable non-viral carriers for gene therapy to treat pathologies affecting the retina. In this study poly(2-(N,N-dimethylamino)ethyl methacrylate), PDMAEMA was synthesized by reversible addition-fragmentation chain transfer (RAFT) and the in vitro cytocompatibility and transfection efficiency of a range of polymer:DNA ratios evaluated using a retinal cell line; in vivo biocompatibility was evaluated by ocular injection in C57BL/6 mice. The results showed that through RAFT, it is possible to produce a defined-size polymer that is compatible with cell viability in vitro and capable of efficiently directing gene expression in a polymer-DNA ratio-dependent manner. When injected into the eyes of mice, these vectors induced a transient, mild inflammation, characteristic of the implantation of medical devices. These results form the basis of future studies where RAFT-synthesized PDMAEMA will be used to deliver gene expression systems to the retina of mouse models of retinal pathologies. Copyright © 2014 John Wiley & Sons, Ltd.
- Electrostatic repulsion between cucurbit[7]urils can be overcome in [3]pseudorotaxane without adding saltsPublication . Pessêgo, Márcia; Moreira, José; Costa, Ana M. Rosa da; Corrochano, P.; Poblete, F. J.; García-Río, LuísThe host−guest chemistry betweencucurbit[7]uril (CB7) and a series of bolaform (Bn) surfactants with different chain lengths, n = 12−22, was the target of our study. [3]Pseudorotaxanes are formed when the alkyl chain of the bolaform has more than 14 carbon atoms. In these cases, two CB7 molecules can be accommodated between the two head groups of the bolaform without addition of electrolytes to the medium. In the case of a bolaform with 12 carbon atoms, the electrostatic repulsion between the carbonyl groups of the CB7 molecules avoids the threading of a second CB7 molecule yielding a mixed structure formed by a [2]pseudorotaxane and an external host−guest complex. The assembly behavior was investigated using NMR spectroscopy, isothermal titration calorimetry (ITC), and kinetic measurements.