M. Luisa González-Martín

Thermodynamic Analysis of Growth Temperature Dependence in the Adhesion of Candida parapsilosis to Polystyrene

ABSTRACT The purpose of this work was to study the adhesion to polystyrene of two Candida parapsilosis strains, grown at 22 and 37°C, in terms of hydrophobicity, surface charge, and interaction free energy. Growth temperature changed the surface properties of microorganisms, yielding a good correlation between thermodynamic predictions and adhesion behavior.

Serum as a Factor Influencing Adhesion of Enterococcus faecalis to Glass and Silicone

ABSTRACT The purpose of this work was to analyze the effect of serum on the physicochemical surface properties and adhesion to glass and silicone of Enterococcus faecalis ATCC 29212 at 37°C. As is presented using thermodynamics analysis, serum minimizes the interaction of cells with water, which correlates well with the increase in hydrophobicity and in bacterial adhesion to glass and silicone.

Atomic force microscopy of mechanically trapped bacterial cells.

This article presents a study on the influence of the protocol used for immobilization of bacterial cells onto surfaces by mechanically trapping them into a filter. In this sense, the surface and structure of trapped cells are analyzed. Bacteria can be present solely or with extracellular polymeric substances (EPS). To test the behavior of the EPS layer duing the filtering process, different strains of a well-known EPS-producer bacteria (Staphylococcus epidermidis), which produce an extracellular matrix clearly visible in AFM images, have been used. Results show that this immobilization method can cause severe structural and mechanical deformation to the cell membrane. This altered mechanical state may possibly influence the parameters derived from AFM force curves (which are micro/nano-mechanical tests). Also, our results suggest that the EPS layer might move during the filtering process and could accumulate at the upper part of the cell, thus favoring distorted data of adhesion/pull-off forces as measured by an AFM tip, especially in the case of submicron-sized microbial cells such as bacteria.

Insights into bacterial contact angles: difficulties in defining hydrophobicity and surface Gibbs energy.

One of the principal techniques for evaluating the surface hydrophobicity of biological samples is contact angle. This method, applied readily to flat-smooth-inert surfaces, gives rise to an important debate when implemented with microbial lawns. After its initial description, in 1984, several authors have carried out modifications of the technique but the results obtained have not been previously judged. This work focuses on the particularities of contact angle measurements on bacterial lawns and enhances the idea of the impossibility of using water contact angle as a universal measurement of bacterial hydrophobicity. Contact angles can only be used as relative indicators of hydrophobicity, in a similar way to tests based on microbial adhesion to solvents. The strong dependence of contact angles on dried bacterial lawns with measuring time and environmental conditions (e.g. pH of the media) preclude the estimation of their absolute values, and so, of the cells surface Gibbs energy. Yet, for a given measuring time, it is found that the hydrophobicity and the apparent bacterial surface Gibbs energy components are qualitatively related to the bacterial surface electrical potential. In particular, a hydrophobic increase is always accompanied by an increase of the cells Lifshitz-Van der Waals component and a decrease of their acid-base component and absolute zeta potential. Therefore, the present study shows that the physico-chemical surface properties that characterize bacteria are not independent, and one of them can be qualitatively described in terms of the others when measuring contact angles at a fixed time after the drying of the microbial beds.

Dendronized Anionic Gold Nanoparticles: Synthesis, Characterization, and Antiviral Activity.

Anionic carbosilane dendrons decorated with sulfonate functions and one thiol moiety at the focal point have been used to synthesize water-soluble gold nanoparticles (AuNPs) through the direct reaction of dendrons, gold precursor, and reducing agent in water, and also through a place-exchange reaction. These nanoparticles have been characterized by NMR spectroscopy, TEM, thermogravimetric analysis, X-ray photoelectron spectroscopy (XPS), UV/Vis spectroscopy, elemental analysis, and zeta-potential measurements. The interacting ability of the anionic sulfonate functions was investigated by EPR spectroscopy with copper(II) as a probe. Different structures and conformations of the AuNPs modulate the availability of sulfonate and thiol groups for complexation by copper(II). Toxicity assays of AuNPs showed that those produced through direct reaction were less toxic than those obtained by ligand exchange. Inhibition of HIV-1 infection was higher in the case of dendronized AuNPs than in dendrons.

Surface-dependent mechanical stability of adsorbed human plasma fibronectin on Ti6Al4V: domain unfolding and stepwise unraveling of single compact molecules.

In this study, the structure and mechanical stability of human plasma fibronectin (HFN), a major protein component of blood plasma, have been evaluated in detail upon adsorption on the nonirradiated and irradiated Ti6Al4V material through the use of atomic force microscopy. The results indicated that the material surface changes occurring after the irradiation process reduce the disulfide bonds that typically preclude the mechanical denaturation of individual HFN domains and interfere significantly with the intraionic interactions stabilizing the compact conformation of the adsorbed HFN molecules. In particular, upon adsorption on this material, the molecules adopt a more flexible conformation and become mechanically more compliant. Unexpected observations also indicated that, regardless the material surface, a single HFN molecule can be pulled into an extended conformation without the unfolding of its domains through a series of three unraveling steps. The forces involved in the unraveling process were found to be generally lower than the forces required to unfold the individual protein domains. This report is the first one to present the force displacement details associated to the straightening of a single compact protein at the molecular level.

Changes on the physico-chemical surface properties and adhesion behaviour of Enterococcus faecalis by the addition of serum or urine to the growth medium

The initial microbial adhesion of bacteria to different surfaces seems to be mediated by physico-chemical forces and this is the reason why the physico-chemical surface characterisation of bacteria has recently gained interest. In this context, the adhesion of different microorganisms to biological substrata has been described from a physico-chemical point of view, aiming to simulate, as closely as possible, the conditions of interest. On this basis, the objective of this work is to characterise the surface of Enterococcus faecalis ATCC29212 through hydrophobicity, surface free energy and zeta potential at 37°C, when cells grow in Trypticase Soy Broth (TSB) and TSB supplemented with serum or urine. These variations are used to provide a theoretical description of the bacterial adhesion to glass by interaction free energy, which is verified with experimental results employing a parallel plate flow chamber set at 37°C, to simulate the conditions of flow inside the human body. The results show that the addition of serum to the growth medium increases the hydrophobicity and isoelectric point (i.e.p.) of the microorganisms, and this could indicate an increase in the protein content on the cell surface. However urine does not introduce a change in the above magnitudes. At short separation distances between the cells and the substratum, the interaction free energy predicts a favourable adhesion for serum-grown cells while non-favourable adhesion is expected for control (TSB-grown cells) and urine-grown cells. These results are in agreement with the experimental adhesion data, obtained with the flow chamber.