The ability of surfactants obtained from three Lactobacillus acidophilus strains to inhibit Staphylococcus aureus and S. epidermidis biofilms was evaluated. Their influence was determined on bacterial initial adhesion, biofilm formation and dispersal using MTT-reduction assay, confocal laser scanning microscopy and image PHLIP analysis. The number of adhering S. aureus and S. epidermidis cells after a 3-h co-incubation with biosurfactants was reduced by 5-56 % in a strain-and dose-dependent manner. S. epidermidis-and, to a lower extent, in S. aureus-biofilm formation was also inhibited in the presence of the tested surfactants. The addition of surfactants to preformed mature biofilms accelerated their dispersal, and changed the parameters of biofilm morphology. The L. acidophilus-derived surfactants inhibit bacterial deposition rate and biofilm development (and also its maturation) without affecting cell growth probably due to the influence on the cell-surface hydrophobicity of staphylococci.

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The aim of the study was to examine the antibiofilm activity of selected essential oils (EO): Lavandula angustifblia (LEO), Melaleuca alternifolia (TTO), Melissa officinalis (MEO) and some of their major constituents: linalool, linalyl acetate, alpha-terpineol, terpinen-4-ol. Biofilms were formed by Staphylococcus aureus ATCC 29213 and Escherichia coli NCTC 8196 on the surface of medical biomaterials (urinary catheter, infusion tube and surgical mesh). TTC reduction assay was used for the evaluation of mature biofilm eradication from these surfaces. Moreover, time-dependent eradication ofbiofilms preformed in polystyrene 96-well culture microplates was examined and expressed as minimal biofilm eradication concentration (evaluated by MTT reduction assay). TTO, alpha-terpineol and terpinen-4-ol as well as MEO, showed stronger anti-biofilm activity than LEO and linalool or linalyl acetate. Among the biomaterials tested, surgical mesh was the surface most prone to persistent colonization since biofilms formed on it, both by S. aureus and E. coli, were difficult to destroy. The killing rate studies of S. aureus biofilm treated with TTO, LEO, MEO and some of their constituents revealed that partial (50%) destruction of 24-h-old biofilms (MBEC50) was achieved by the concentration 4-8 x MIC after 1 h, whereas 2-4 x MIC was enough to obtain 90% reduction in biomass metabolic activity (MBEC90) after just 4 h of treatment. A similar dose-dependent effect was observed for E. coli biofilm which, however, was more susceptible to the action of phytochemicals than the biofilms of S. aureus. It is noteworthy that an evident decrease in biofilm cells metabolic activity does not always lead to their total destruction and eradication.

Titania nanotube (TNT) coatings were produced using low-potential anodic oxidation of Ti6Al4V substrates in the potential range 3–20 V. They were analysed by X-ray diffraction (XRD), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS), and scanning electron microscopy (SEM). The wettability was estimated by measuring the contact angle when applying water droplets. The bioactivity of the TNT coatings was established on the basis of the biointegration assay (L929 murine fibroblasts adhesion and proliferation) and antibacterial tests against Staphylococcus aureus (ATCC 29213). The photocatalytic efficiency of the TNT films was studied by the degradation of methylene blue under UV irradiation. Among the studied coatings, the TiO2 nanotubes obtained with the use of 5 V potential (TNT5) were found to be the most appropriate for medical applications. The TNT5 sample possessed antibiofilm properties without enriching it by additional antimicrobial agent. Furthermore, it was characterized by optimal biocompatibility, performing better than pure Ti6Al4V alloy. Moreover, the same sample was the most photocatalytically active and exhibited the potential for the sterilization of implants with the use of UV light and for other environmental applications.

The colonization of respiratory tract by Staphylococcus aureus is a frequent feature of cystic fibrosis (CF), especially in pediatric patients. The formation of small colony variants (SCVs), which produce reduced amounts of alpha-toxin, is one of the proposed ways of staphylococcal accommodation in an intracellular niche. The aim of the present study was to compare some properties of S. aureus SCVs and their parent strains. A site-directed S. aureus hemB mutant and parent strain 8325-4 were included in the study (control pair). Normal and SCV strain pairs from CF patients as well as control strains were tested for the susceptibility to defensins, killing activity of professional phagocytes and adhesion to A549 cell line. Because S. aureus are exposed to many cationic proteins in the host, we challenged a clinical isolate with minimal subinhibitory concentration (subMIC) of protamine and found that hemin and menadione auxotrophic SCVs emerged. SCVs were more resistant than normal strains to protamine but not to dermaseptin. The susceptibility to the bactericidal activity of magainin was the same for normal and SCV strains. The protamine resistance of normal as well as SCVs was strongly enhanced by high salt concentration. The adhesion of some SCVs to A549 cells was higher than adhesion of parental strains. However, the number of adherent bacteria (SCVs) was diminished in the presence of hemin for hemin auxotrophs. The uptake of SCVs by granulocytes was lower than ingestion of normal strains, but SCVs were killed with equal or greater potency. SCVs are adapted to intracellular survival and persistence in the host under certain circumstances. The ability to form a variant subpopulation affords S. aureus additional survival options.

New antimicrobial properties of products derived from Humulus lupulus L. such as antiadherent and antibiofilm activities were evaluated. The growth of gram-positive but not gram-negative bacteria was inhibited to different extents by these compounds. An extract of hop cones containing 51% xanthohumol was slightly less active against S. aureus strains (MIC range 31.2–125.0 μg/mL) than pure xanthohumol (MIC range 15.6–62.5 μg/mL). The spent hop extract, free of xanthohumol, exhibited lower but still relevant activity (MIC range 1-2 mg/mL). There were positive coactions of hop cone, spent hop extracts, and xanthohumol with oxacillin against MSSA and with linezolid against MSSA and MRSA. Plant compounds in the culture medium at sub-MIC concentrations decreased the adhesion of Staphylococci to abiotic surfaces, which in turn caused inhibition of biofilm formation. The rate of mature biofilm eradication by these products was significant. The spent hop extract at MIC reduced biofilm viability by 42.8%, the hop cone extract by 74.8%, and pure xanthohumol by 86.5%. When the hop cone extract or xanthohumol concentration was increased, almost complete biofilm eradication was achieved (97–99%). This study reveals the potent antibiofilm activity of hop-derived compounds for the first time.

The highly ordered titanium dioxide nanotube coatings were produced under various electrochemical conditions on the surface of titanium foil. The anodization voltage changes proved to be a main factor which directly affects the nanotube morphology, structure, and wettability. Moreover we have noticed a significant dependence between the size and crystallinity of TiO2 layers and the adhesion/proliferation of fibroblasts and antimicrobial properties. Cellular functionality were investigated for up to 3 days in culture using a cell viability assay and scanning electron microscopy. In general, results of our studies revealed that fibroblasts adhesion, proliferation, and differentiation on the titania nanotube coatings is clearly higher than on the surface of the pure titanium foil. The formation of crystallic islands in the nanotubes structure induced a significant acceleration in the growth rate of fibroblasts cells by as much as ~200 %. Additionally, some types of TiO2 layers revealed the ability to the reduce of the staphylococcal aggregates/biofilm formation. The nanotube coatings formed during the anodization process using the voltage 4 V proved to be the stronger S. aureus aggregates/biofilm inhibitor in comparison to the uncovered titanium substrate. That accelerated eukaryotic cell growth and anti-biofilm activity is believed to be advantageous for faster cure of dental and orthopaedic patients, and also for a variety of biomedical diagnostic and therapeutic applications.Graphical AbstractThe highly ordered titanium dioxide nanotube coatings were produced under various electrochemical conditions on the surface of titanium foil. The anodization voltage changes proved to be a main factor which directly affects the nanotube morphology, structure, and wettability. Moreover we have noticed a significant dependence between the size and crystallinity of TiO2 layers and the adhesion/proliferation of fibroblasts and antimicrobial properties.

JC virus (JCV) causes a fatal demyelinating disease, progressive multifocal leukoencephalopathy, in immunocompromised patients (4). The promoter and enhancer elements present within the regulatory region of the JCV prototype Mad-1 strain consist of two 98-bp tandem repeats, each of which contains a partially characterized binding site for activating transcription factor 1 (AP-1) adjacent to the NF-1 binding element at nucleotide positions 56 to 63 and 154 to 161 (12, 24, 39). An additional AP-1-like site is also juxtaposed to an NF-1 site at positions 262 to 269 (39) (see Fig. 1A). Although earlier studies revealed the interaction of an AP-1 family member, c-Jun, with this binding site on the JCV promoter (1), its involvement in JCV gene expression remained unclear.AP-1 was first defined as an inducible DNA binding protein specific to positive regulatory elements found within the regulatory regions of both simian virus 40 (SV40) and the metallothionein gene (2,17,18). Biochemical purification showed that AP-1 is not a single transcription factor but instead a series of related dimeric complexes of the Jun (c-Jun, JunB, and JunD) and Fos (c-Fos, FosB, Fra-1, and Fra-2) families (3, 42). Each family member is a phosphonuclear protein composed of three distinct functional domains, including a carboxy-terminal leucine zipper domain followed by an adjacent basic domain and an amino-terminal transactivation domain. AP-1 family members are induced by a wide variety of signals, including, but not limited to, UV and ionizing radiation, oxidative stress, neuronal depolarization, cytokines (tumor necrosis factor alpha, gamma interferon, and interleukin 1), and viral infection (6, 11, 13-15, 31, 32, 44). The AP-1 family members are collectively known as proto-oncogenes because of their high sequence homology to some retrovirus-encoded oncogenic proteins and their involvement in many cellular processes, including cell proliferation, survival, and apoptosis (5,19,31,38,42).In light of earlier data on the activation of AP-1 upon various viral infections (10,16,20,30,43) and the fact that JCV promoter-enhancer elements contain AP-1 binding sites, we investigated the effect of AP-1 on JCV gene regulation and obtained experimental evidence that the AP-1 family members c-Jun and c-Fos positively regulate transcription from both JCV early and late promoters. On the other hand, we assessed the effect of JCV infection on c-Jun expression levels and showed that c-Jun protein levels are modulated and modified by phosphorylation during the viral infection cycle.c-Jun interacts with potential AP-1 binding site from JCV promoter in vitro. Upon induction by a variety of extracellular stimuli, the AP-1 family of transcription factors bind to their cognate DNA sequences and stimulate transcription of AP-1-responsive promoters (3,23,35). UV irradiation-mediated stress is known to be one of the potent inducers of the AP-1 family of transcription factors (3, 31). Since JCV promoterenhancer elements contain several potential binding sites for A...