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The mechanism of action of pepR, a viral-derived peptide, against Staphylococcu...

Pinto, Sandra; Dias, Susana; Cruz, Ana F; Mil-Homens, Dalila; Fernandes, Fabio; Valle, Javier; Andreu, David; Prieto, Manuel; Castanho, Miguel A. R. B.

Objectives: To investigate the mechanism of action at the molecular level of pepR, a multifunctional peptide derived from the Dengue virus capsid protein, against Staphylococcus aureus biofilms. Methods: Biofilm mass, metabolic activity and viability were quantified using conventional microbiology techniques, while fluorescence imaging methods, including a real-time calcein release assay, were employed to inves...


FRET in Membrane Biophysics: An Overview

Loura, Luís M. S.; Prieto, Manuel

Förster resonance energy transfer (FRET), in most applications used as a "spectroscopic ruler," allows an easy determination of the donor-acceptor intermolecular distance. However, the situation becomes complex in membranes, since around each donor there is an ensemble of acceptors at non-correlated distances. In this review, state-of-the-art methodologies for this situation are presented, usually involving tim...


Effect of ionic strength and presence of serum on lipoplexes structure monitori...

Madeira, Catarina; Loura, Luís M. S.; Prieto, Manuel; Fedorov, Aleksander; Aires-Barros, M. Raquel

Background: Serum and high ionic strength solutions constitute important barriers to cationic lipid-mediated intravenous gene transfer. Preparation or incubation of lipoplexes in these media results in alteration of their biophysical properties, generally leading to a decrease in transfection efficiency. Accurate quantification of these changes is of paramount importance for the success of lipoplex-mediated gen...


Interaction of S413-PV cell penetrating peptide with model membranes: relevance...

Mano, Miguel; Henriques, Ana; Paiva, Artur; Prieto, Manuel; Gavilanes, Francisco; Simões, Sérgio; Lima, Maria C. Pedroso De

Cell penetrating peptides (CPPs) have been successfully used to mediate the intracellular delivery of a wide variety of molecules of pharmacological interest both in vitro and in vivo, although the mechanisms by which the cellular uptake occurs remain unclear and controversial. Following our previous work demonstrating that the cellular uptake of the S413-PV CPP occurs mainly through an endocytosis-independent ...


Ciprofloxacin interactions with bacterial protein OmpF: Modelling of FRET from ...

Fernandes, Fábio; Neves, Patrícia; Gameiro, Paula; Loura, Luís M. S.; Prieto, Manuel

The outer membrane protein F (OmpF) is known to play an important role in the uptake of fluoroquinolone antibiotics by bacteria. In this study, the degree of binding of the fluoroquinolone antibiotic ciprofloxacin to OmpF in a lipid membrane environment is quantified using a methodology based on Förster resonance energy transfer (FRET). Analysis of the fluorescence quenching of OmpF is complex as each OmpF mono...


The 9-antroate chromophore as a fluorescence probe for water

Costa, Fernando Pestana da; Costa, Sílvia B.; Melo, Eurico C.; Prieto, Manuel; Maçanita, António L.


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