Staphylococcus aureus-Fibronectin Interactions with and without Fibronectin-Binding Proteins and Their Role in Adhesion and Desorption

Adhesion and residence-time-dependent desorption of two Staphylococcus aureus strains with and without fibronectin (Fn) binding proteins (FnBPs) on Fn-coated glass were compared under flow conditions. To obtain a better understanding of the role of Fn-FnBP binding, the adsorption enthalpies of Fn with staphylococcal cell surfaces were determined using isothermal titration calorimetry (ITC). Interaction forces between staphylococci and Fn coatings were measured using atomic force microscopy (AFM). The strain with FnBPs adhered faster and initially stronger to an Fn coating than the strain without FnBPs, and its Fn adsorption enthalpies were higher. The initial desorption was high for both strains but decreased substantially within 2 s. These time scales of staphylococcal bond ageing were confirmed by AFM adhesion force measurement. After exposure of either Fn coating or staphylococcal cell surfaces to bovine serum albumin (BSA), the adhesion of both strains to Fn coatings was reduced, suggesting that BSA suppresses not only nonspecific but also specific Fn-FnBP interactions. Adhesion forces and adsorption enthalpies were only slightly affected by BSA adsorption. This implies that under the mild contact conditions of convective diffusion in a flow chamber, adsorbed BSA prevents specific interactions but does allow forced Fn-FnBP binding during AFM or stirring in ITC. The bond strength energies calculated from retraction force-distance curves from AFM were orders of magnitude higher than those calculated from desorption data, confirming that a penetrating Fn-coated AFM tip probes multiple adhesins in the outermost cell surface that remain hidden during mild landing of an organism on an Fn-coated substratum, like that during convective diffusional flow

Evaluation of adhesion forces of Staphylococcus aureus along the length of Candida albicans hyphae

Background Candida albicans is a human fungal pathogen, able to cause both superficial and serious, systemic diseases and is able to switch from yeast cells to long, tube-like hyphae, depending on the prevailing environmental conditions. Both morphological forms of C. albicans are found in infected tissue, often in combination with Staphylococcus aureus. Although bacterial adhesion to the different morphologies of C. albicans has been amply studied, possible differences in staphylococcal adhesion forces along the length of C. albicans hyphae have never been determined. In this study, we aim to verify the hypothesis that the forces mediating S. aureus NCTC8325-4GFP adhesion to hyphae vary along the length of C. albicans SC5314 and MB1 hyphae, as compared with adhesion to yeast cells. Results C. albicans hyphae were virtually divided into a "tip" (the growing and therefore youngest part of the hyphae), a "middle" and a so-called "head" region (the yeast cell from which germination started). Adhesion forces between S. aureus NCTC8325-4GFP and the different regions of C. albicans SC5314 hyphae were measured using atomic force microscopy. Strong adhesion forces were found at the tip and middle regions of C. albicans hyphae (−4.1 nN and −4.0 nN, respectively), while much smaller adhesion forces were measured at the head region (−0.3 nN). Adhesion forces exerted by the head region were comparable with the forces arising from budding yeast cells (−0.5 nN). A similar regional dependence of the staphylococcal adhesion forces was found for the clinical isolate involved in this study, C. albicans MB1. Conclusions This is the first time that differences in adhesion forces between S. aureus and different regions of C. albicans hyphae have been demonstrated on a quantitative basis, supporting the view that the head region is different from the remainder of the hyphae. Notably it can be concluded that the properties of the hyphal head region are similar to those of budding yeast cells. These novel findings provide new insights in the intricate interkingdom interaction between C. albicans and S. aureus

The use of positively charged or low surface free energy coatings versus polymer brushes in controlling biofilm formation

Biofilm formation on biomaterials implant surfaces and subsequent infectious complications are a frequent reason for failure of many biomedical devices, such as total hip arthroplasties, vascular catheters and urinary catheters. The development of a biofilm is initiated by the formation of a conditioning film of adsorbed macromolecules, such as proteins, followed by adhesion of microorganisms, where after they grow and anchor through secretion of extracellular polymeric substances. Adhesion of microorganisms is influenced by the physico-chemical properties of the biomaterial surface. Positively charged materials stimulate bacterial adhesion, but prevent growth of adhering bacteria. The use of low surface free energy materials did not always reduce in vitro adhesion of bacteria, but has been found beneficial in in vivo applications where fluctuating shear forces prevail, like on intra-oral devices and urine catheters. Polymer brushes have shown a very high reduction in in vitro adhesion of a great variety of microorganisms. However, for clinical application, the long term stability of polymer brushes is still a limiting factor. Further effort is therefore required to enhance the stability of polymer brushes on biomaterial implant surfaces to facilitate clinical use of these promising coating

The influence of ionic strength on the adhesive bond stiffness of oral streptococci possessing different surface appendages as probed using AFM and QCM-D

Bacterial adhesion to surfaces poses threats to human-health, not always associated with adhering organisms, but often with their detachment causing contamination elsewhere. Bacterial adhesion mechanisms may not be valid for their detachment, known to proceed according to a visco-elastic mechanism. Here we aimed to investigate influences of ionic strength on the adhesive bond stiffness of two spherically shaped Streptococcus salivarius strains with different lengths of fibrillar surface appendages. The response of a Quartz-Crystal-Microbalance-with-Dissipation (QCM-D) upon streptococcal adhesion and changes in the ionic strength of the surrounding fluid indicated that the bond stiffness of S. salivarius HB7, possessing a dense layer of 91 nm long fibrils, was unaffected by ionic strength. Atomic-force-microscopic (AFM) imaging in PeakForce-QNM mode showed a small decrease in bond stiffness from 1200 to 880 kPa upon decreasing ionic strength from 57 to 5.7 mM, while Total-Internal-Reflection-Microscopy suggested a complete collapse of fibrils. S. salivarius HBV51, possessing a less dense layer of shorter (63 nm) fibrils, demonstrated a strong decrease in bond stiffness both from QCM-D and AFM upon decreasing the ionic strength, and a partial collapse of fibrils. Probably, the more hydrophobic and less negatively charged long fibrils on S. salivarius HB7 collapse side-on to the cell surface, while the more hydrophilic and negatively charged fibrils of S. salivarius HBV51 remain partially stretched. In summary, we demonstrate how a combination of different methods can yield a description of the structural changes occurring in the interfacial region between adhering, fibrillated streptococci and a substratum surface upon changing the ionic strengt

Relations between macroscopic and microscopic adhesion of Streptococcus mitis strains to surfaces

Application of physico-chemical models to describe bacterial adhesion to surfaces has hitherto only been partly successful due to the structural and chemical heterogeneities of bacterial surfaces, which remain largely unaccounted for in macroscopic physico-chemical characterizations of the cell surfaces. In this study, the authors attempted to correlate microscopic adhesion of a collection of nine Streptococcus mitis strains to the negatively charged, hydrophilic silicon nitride tip of an atomic force microscope (AFM) with macroscopic adhesion of the strains to a negatively charged, hydrophilic glass in a parallel-plate flow chamber. The repulsive force probed by AFM upon approach of the tip to a bacterial cell surface ranged from 1·7 to 7·7 nN depending on the strain considered and was found to correspond to an activation barrier, governing initial, macroscopic adhesion of the organisms to the glass surface. Moreover, maximum distances at which attractive forces were probed by the AFM upon retraction of the tip (120 to 1186 nm) were related to the area blocked by an adhering bacterium, i.e. the distance kept between adhering bacteria. Bacterial desorption could not be related to adhesive forces as probed by the AFM, possibly due to the distinct nature of the desorption process occurring in the parallel-plate flow chamber and the forced detachment in AFM

Inhibition of adhesion of yeasts and bacteria by poly(ethylene oxide-)brushes on glass in a parallel plate flow chamber

Poly(ethylene oxide) (PEO)-brushes are generally recognized as protein-repellent surfaces, and although a role in discouraging microbial adhesion has been established for some strains and species, no study exists on the effects of PEO-brushes on a large variety of bacterial and yeast strains. In this paper, a PEO-brush has been covalently attached to glass and silica by reaction in a polymer melt. Subsequently, the presence of a PEO-brush was demonstrated using contact angle measurements, X-ray photoelectron spectroscopy and ellipsometry. For five bacterial (Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus salivarius, Escherichia coli and Pseudomonas aeruginosa) and two yeast strains (Candida albicans and Candida tropicalis), adhesion to PEO-brushes was compared with adhesion to bare glass in a parallel plate flow chamber. The initial deposition rates of Sta. epidermidis, Sta. aureus and Str. salivarius to glass were relatively high, between 2400 and 2600 cm-2 s-1, while E. coli and P. aeruginosa deposited much more slowly. The initial deposition rates of the yeasts to glass were 144 and 444 cm-2 s-1 for C. albicans GB 1/2 and C. tropicalis GB 9/9, respectively. Coating of the glass surface with a PEO-brush yielded more than 98 % reduction in bacterial adhesion, although for the more hydrophobic P. aeruginosa a smaller reduction was observed. For both yeast species adhesion suppression was less effective than for the bacteria and here too the more hydrophobic C. tropicalis showed less reduction than the more hydrophilic C. albicans. The PEO-brush had a thickness of 22 nm in water, as inferred from ellipsometry. Assuming that on bare glass the adhered micro-organisms are positioned only a few nanometers away from the surface and that the brush keeps them at a distance of 22 nm, it is calculated that the brush yields a sevenfold attenuation of the Lifshitz–Van der Waals attraction to the surface between the micro-organisms and the surface. Decreased Lifshitz–van der Waals attraction may be responsible for the suppression of the microbial adhesion observed

Reduced waiting times by preference-based allocation of patients to nursing homes

Objectives: The long waiting times for nursing homes can be reduced by applying advanced waiting-line management. In this article, we implement a preference-based allocation model for older adults to nursing homes, evaluate the performance in a simulation setting for 2 case studies, and discuss the implementation in practice.Design: Simulation study.Setting and Participants: Older adults requiring somatic nursing home care, from an urban region (Rotterdam) and a rural region (Twente) in the Netherlands.Methods: Data about nursing homes and capacities for the 2 case studies were identified. A set of preference profiles was defined with aims regarding waiting time preferences and flexibility. Guidelines for implementation of the model in practice were obtained by addressing the tasks of all stakeholders. Thereafter, the simulation was run to compare the current practice with the allocation model based on specified outcome measures about waiting times and preferences.Results: We found that the allocation model decreased the waiting times in both case studies. Compared with the current practice policy, the allocation model reduced the waiting times until placement by at least a factor of 2 (from 166 to 80 days in Rotterdam and 178 to 82 days in Twente). Moreover, more of the older adults ended up in their preferred nursing home and the aims of the distinct preference profiles were satisfied.Conclusions and Implications: The results show that the allocation model outperforms commonly used waiting-line policies for nursing homes, while meeting individual preferences to a larger extent. Moreover, the model is easy to implement and of a generic nature and can, therefore, be extended to other settings as well (eg, to allocate older adults to home care or daycare). Finally, this research shows the potential of mathematical models in the care domain for older adults to face the increasing need for cost-effective solutions.</p

The Planetary Nebula Luminosity Function at the Dawn of Gaia

The [O III] 5007 Planetary Nebula Luminosity Function (PNLF) is an excellent extragalactic standard candle. In theory, the PNLF method should not work at all, since the luminosities of the brightest planetary nebulae (PNe) should be highly sensitive to the age of their host stellar population. Yet the method appears robust, as it consistently produces < 10% distances to galaxies of all Hubble types, from the earliest ellipticals to the latest-type spirals and irregulars. It is therefore uniquely suited for cross-checking the results of other techniques and finding small offsets between the Population I and Population II distance ladders. We review the calibration of the method and show that the zero points provided by Cepheids and the Tip of the Red Giant Branch are in excellent agreement. We then compare the results of the PNLF with those from Surface Brightness Fluctuation measurements, and show that, although both techniques agree in a relative sense, the latter method yields distances that are ~15% larger than those from the PNLF. We trace this discrepancy back to the calibration galaxies and argue that, due to a small systematic error associated with internal reddening, the true distance scale likely falls between the extremes of the two methods. We also demonstrate how PNLF measurements in the early-type galaxies that have hosted Type Ia supernovae can help calibrate the SN Ia maximum magnitude-rate of decline relation. Finally, we discuss how the results from space missions such as Kepler and Gaia can help our understanding of the PNLF phenomenon and improve our knowledge of the physics of local planetary nebulae.Comment: 12 pages, invited review at the conference "The Fundamental Cosmic Distance Scale: State of the Art and Gaia Perspective", to appear in Astrophysics and Space Scienc

Planck 2015 results. XXVII. The Second Planck Catalogue of Sunyaev-Zeldovich Sources

We present the all-sky Planck catalogue of Sunyaev-Zeldovich (SZ) sources detected from the 29 month full-mission data. The catalogue (PSZ2) is the largest SZ-selected sample of galaxy clusters yet produced and the deepest all-sky catalogue of galaxy clusters. It contains 1653 detections, of which 1203 are confirmed clusters with identified counterparts in external data-sets, and is the first SZ-selected cluster survey containing > $10^3$ confirmed clusters. We present a detailed analysis of the survey selection function in terms of its completeness and statistical reliability, placing a lower limit of 83% on the purity. Using simulations, we find that the Y5R500 estimates are robust to pressure-profile variation and beam systematics, but accurate conversion to Y500 requires. the use of prior information on the cluster extent. We describe the multi-wavelength search for counterparts in ancillary data, which makes use of radio, microwave, infra-red, optical and X-ray data-sets, and which places emphasis on the robustness of the counterpart match. We discuss the physical properties of the new sample and identify a population of low-redshift X-ray under- luminous clusters revealed by SZ selection. These objects appear in optical and SZ surveys with consistent properties for their mass, but are almost absent from ROSAT X-ray selected samples