In the human fungal pathogen Candida albicans, the CUG codon is translated 97% of the time as serine and 3% of the time as leucine, which potentially originates an array of proteins resulting from the translation of a single gene. Genes encoding cell surface proteins are enriched in CUG codons; thus, CUG mistranslation may influence the interactions of the organism with the host. To investigate this, we compared a C. albicans strain that misincorporates 28% of leucine at CUGs with a wild-type parental strain. The first strain displayed increased adherence to inert and host molecules. In addition, it was less susceptible to phagocytosis by murine macrophages, probably due to reduced exposure of cell surface β-glucans. To prove that these phenotypes occurred due to serine/leucine exchange, the C. albicans adhesin and invasin ALS3 was expressed in Saccharomyces cerevisiae in its two natural isoforms (Als3p-Leu and Als3p-Ser). The cells with heterologous expression of Als3p-Leu showed increased adherence to host substrates and flocculation. We propose that CUG mistranslation has been maintained during the evolution of C. albicans due to its potential to generate cell surface variability, which significantly alters fungus-host interactions.

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Biofilms are commonly involved in medical device-related infections. The purpose of this study was to determine the antimicrobial and anti-biofilm activity of polyethyleneimine (PEI) and PEIbased nanoparticles (nanoPEI) against Staphylococcus aureus, Staphylococcus epidermidis, Acinetobacter baumannii and Candida albicans (clinical and ATCC strains), and to evaluate their effect upon biofilm formation on polyurethane (PUR)-like catheters. MICs and minimal lethal concentrations of PEI and nanoPEI were determined according to CLSI microdilution reference protocols. For PEI, the MIC value was 195.31 mg l "1 for all the bacteria and 48.83 mg l "1 for the yeast strains. For nanoPEI, the MIC value was 1250 mg l "1 for all the strains except A. baumannii, for which it was 2500 mg l "1. Biofilm formation was assessed with PUR-like catheter segments and biofilm metabolic activity was quantified by colorimetry with a tetrazolium reduction assay. Plasma membrane integrity and membrane potential were assessed by flow cytometry after staining microbial cells with a membrane-impermeable dye, propidium iodide, and a membranepotential marker, DiBAC 4 (3). PEI inhibited growth of all microbial species; higher concentrations of nanoPEI were needed to inhibit growth of all species. Biofilm formation in the presence of antibacterial PEI activity was dose-dependent (except for S. epidermidis) and species-related. NanoPEI at 0.5¾MIC and MIC significantly reduced the metabolic activity of biofilms of S. aureus, S. epidermidis and A. baumannii, whereas 2¾MIC was required in order to inhibit biofilm metabolic activity.

During the last 30 years the incidence of fungal infections has increased dramatically. While the antifungal therapeutic options available are somewhat reduced, most pathogenic microorganisms have an incredible capacity to mutate and acquire resistance. In addition, multiple drugs are often required concomitantly to manage clinically complex disorders. The combination of antibiotics or other compounds with antifungal drugs, simultaneously or sequentially, is commonly adopted in clinical practice, although without a full knowledge of the consequences. Thus, the role of combined therapy and the effect of antibiotics upon fungal growth promotion need to be critically evaluated and understood in order to avoid undesirable drug interactions. With this review we intend discuss the studies that report about antibiotics inhibiting fungal growth, as well as studies describing the synergistic effect of the combined therapy, i.e., associations between antibiotics or other compounds with antifungal drugs. Alternative therapeutic protocols for fungal disease could be designed, taking advantage of such drug combinations. Critical revision of previously published data is crucial in order to define future research strategies.

Several studies have explored the antimicrobial properties of manuka honey (MkH). However, the data available regarding antibacterial action mechanisms are scarcer. The aim of this study was to scrutinize and characterize primary effects of manuka honey (MkH) upon the physiological status of Staphylococcus aureus and Escherichia coli (as Gram-positive and Gram-negative bacteria models, respectively), using flow cytometry (FC) to reveal its antibacterial action mechanisms. Effects of MkH on membrane potential, membrane integrity and metabolic activity were assessed using different fluorochromes in a 180 min time course assay. Time-kill experiments were carried out under the same conditions. Additionally, MkH effect on efflux pumps was also studied in an E. coli strain with an over-expression of several efflux pumps. Exposure of bacteria to MkH resulted in physiological changes related to membrane potential and membrane integrity; these effects displayed slight differences among bacteria. MkH induced a remarkable metabolic disruption as primary physiological effect upon S. aureus and was able to block efflux pump activity in a dose-dependent fashion in the E. coli strain.

Medical device-associated infections (MDAI) are a critical problem due to the increasing usage of medical devices in the aging population. The inhibition of biofilm formation through the use of probiotics has received attention from the medical field in the last years. However, this sparse knowledge has not been properly reviewed, so that successful strategies for biofilm management can be developed. This study aims to summarize the relevant literature about the effect of probiotics and their metabolites on biofilm formation in medical devices using a PRISMA-oriented (Preferred Reporting Items for Systematic reviews and Meta-Analyses) systematic search and meta-analysis. This approach revealed that the use of probiotics and their products is a promising strategy to hinder biofilm growth by a broad spectrum of pathogenic microorganisms. The meta-analysis showed a pooled effect estimate for the proportion of biofilm reduction of 70% for biosurfactants, 76% for cell-free supernatants (CFS), 77% for probiotic cells and 88% for exopolysaccharides (EPS). This review also highlights the need to properly analyze and report data, as well as the importance of standardizing the in vitro culture conditions to facilitate the comparison between studies. This is essential to increase the predictive value of the studies and translate their findings into clinical applications.

Understanding the conditions affecting cyanobacterial biofilm development is crucial to develop new antibiofouling strategies and decrease the economic and environmental impact of biofilms in marine settings. In this study, we investigated the relative importance of shear forces and surface hydrophobicity on biofilm development by two coccoid cyanobacteria with different biofilm formation capacities. The strong biofilm-forming Synechocystis salina was used along with the weaker biofilm-forming Cyanobium sp. Biofilms were developed in defined hydrodynamic conditions using glass (a model hydrophilic surface) and a polymeric epoxy coating (a hydrophobic surface) as substrates. Biofilms developed in both surfaces at lower shear conditions contained a higher number of cells and presented higher values for wet weight, thickness, and chlorophyll a content. The impact of hydrodynamics on biofilm development was generally stronger than the impact of surface hydrophobicity, but a combined effect of these two parameters strongly affected biofilm formation for the weaker biofilm-producing organism. The antibiofilm performance of the polymeric coating was confirmed at the hydrodynamic conditions prevailing in ports. Shear forces were shown to have a profound impact on biofilm development in marine settings regardless of the fouling capacity of the existing flora and the hydrophobicity of the surface.

Several methodologies have been implemented with the intent of preventing or reducing the formation of biofilms on indwelling urinary devices. The use of carbon nanotubes (CNTs) in the biomedical field has been increasing, particularly in the production of antimicrobial and antifouling coatings. Despite their proven antimicrobial properties, their use as coating materials for urinary tract devices (UTDs) is still poorly documented. In the present work, CNT/poly(dimethylsiloxane) (PDMS) composite materials containing different CNT loadings were prepared and further tested against Escherichia coli under conditions prevailing in UTDs. Besides CNT loading optimization, textural modifications were also introduced on the surface of CNTs to improve the antibiofilm pro-perties of the final composites. Material characterization included the textural evaluation of CNTs and the assessment of surface morphology by scanning electron microscopy, while the surface hydrophobicity was determined by contact angle measurements. Biofilm analysis was performed by determining the number of culturable and total cells and by confocal laser scanning microscopy. Results revealed that, by filling the PDMS matrix with 3 wt% CNT loading, a significant reduction in cell culturability (39%) can be achieved compared to PDMS. Additionally, the textural modifications induced by ball-milling treatment proved to be effective on the inhibition of biofilm formation, reducing the amount of biofilm per surface area, biofilm thickness and surface coverage in 31, 47 and 27%, respectively (compared to surfaces where CNTs were not ball-milled).