3D-PLA-experimental set up to display the electrical background of the so-called geometric factor of electrokinetic cells.

The so-called geometric factor defined in electrokinetic cells, L/S (L being the length and S the cross-section of the channel), is relevant for providing the surface interaction electrical potential (zeta potential, ζ) of large surfaces, such as those used in the design of biomedical devices or water purification systems. Conversely, recent studies demonstrate that this factor is also employed to determine geometrical parameters, such as porosity in membrane-like systems. This factor, which has been attributed exclusively a geometrical character, can also be obtained from the electrical conductivity and resistance of the electrokinetic channel. In this work, we assess whether these two ways of obtaining the L/S factor are equivalent and how possible deviations can affect the value of the zeta potential. For this purpose, we work with channels of different geometries obtained by 3D printing using PLA (polylactic acid) as a polymer employed in biomedical applications. The discrepancies between the L/S factor obtained by electrical and purely geometrical measurements increase as the geometrical L/S factor becomes larger, reaching differences close to 80%. The results show that the so-called geometrical L/S factor also has an important electrical contribution and would be better denoted as electrogeometric factor. The differences found between the L/S factors are also propagated to the calculation of ζ but an optimum conductivity zone (from about 10 to 40 mS m-1) can be defined to obtain the zeta potential by selecting any of the L/S factors obtained from electrokinetic measurements. The results of this work should be taken into account in those investigations that use the L/S factor to obtain the geometry-porosity of permeable materials.

Characterization of Magnesium-Polylactic Acid Films Casted on Different Substrates and Doped with Diverse Amounts of CTAB.

Polylactic acid (PLA) is a good candidate for the manufacture of polymeric biodegradable biomaterials. The inclusion of metallic particles and surfactants solves its mechanical limitations and improves its wettability, respectively. In this work, cetyltrimethylammonium bromide (CTAB) and magnesium particles have been incorporated into PLA films to evaluate the changes produced in the polymeric matrix cast on glass and silicone substrates. For this purpose, the surface of the films has been characterized by means of contact angle measurements and ToF-SIMS. Depth profiles and SEM images of the cross sections of the films have also been obtained to study their morphology. The results show that the CTAB in the polymer matrix with and without magnesium improves the wettability of the films, making them more suitable for cell adhesion. The higher the hydrophilicity, the higher the surfactant concentration. The depth profiles show, for the first time, that, depending on the surfactant concentration and the presence of Mg, there is a layer-like distribution near the surface where, in addition to the CTAB + PLA mixture, a surfactant exclusion zone can be seen. This new structure could be relevant in in vitro/in vivo situations when the degradation processes remove the film components in a sequential form.

In vivo bactericidal efficacy of the Ti6Al4V surface after ultraviolet C treatment.

BACKGROUND: Biomaterial-associated infections are one of the most important complications in orthopedic surgery. The main goal of this study was to demonstrate the in vivo bactericidal effect of ultraviolet (UV) irradiation on Ti6Al4V surfaces. MATERIALS AND METHODS: An experimental model of device-related infections was developed by direct inoculation of Staphylococcus aureus into the canal of both femurs of 34 rats. A UV-irradiated Ti6Al4V pin was press-fit into the canal by retrograde insertion in one femur and the control pin was inserted into the contralateral femur. To assess the efficacy of UV radiation, the mean colony counts after inoculation in the experimental subjects and the control group were compared at different times of sacrifice and at different inoculum doses. RESULTS: At 72 h, the mean colony counts after inoculation in experimental femurs were significantly lower than those of the control group, with a reduction percentage of 76 % (p = 0.041). A similar difference between control and experimental pins was observed at 24 h using an inoculum dose <104 colony-forming units (CFU), for which the reduction percentage was 70.48 % (p = 0.017). CONCLUSION: The irradiated surface of Ti6Al4V is able to reduce early bacterial colonization of Ti6AlV pins located in the medullar channel and in the surrounding femur. The reductions depend on the initial inoculums used to cause infection in the animals and the greatest effects are detected for inoculums <104 CFU. LEVEL OF EVIDENCE: Not applicable.

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.

PIK3CA gene alterations in bladder cancer are frequent and associate with reduced recurrence in non-muscle invasive tumors.

Bladder cancer (BC) is the fifth most common cancer in the world, being the non-muscle invasive tumors (NMIBC) the most frequent. NMIBC shows a very high frequency of recurrence and, in certain cases, tumor progression. The phosphatidylinositol 3-kinase (PI3K) pathway, which controls cell growth, tumorigenesis, cell invasion and drug response, is frequently activated in numerous human cancers, including BC, in part through alterations of PIK3CA gene. However, the significance of PIK3CA gene alterations with respect to clinicopathological characteristics, and in particular tumor recurrence and progression, remains elusive. Here, we analyzed the presence of mutations in FGFR3 and PIK3CA genes and copy number alterations of PIK3CA gene in bladder tumor and their correspondent paired normal samples from 87 patients. We observed an extremely high frequency of PIK3CA gene alterations (mutations, copy gains, or both) in tumor samples, affecting primarily T1 and T2 tumors. A significant number of normal tissues also showed mutations and copy gains, being coincident with those found in the corresponding tumor sample. In low-grade tumors PIK3CA mutations associated with FGFR3 mutations. Alterations in PIK3CA gene resulted in increased Akt activity in tumors. Interestingly, the presence of PIK3CA gene alterations, and in particular gene mutations, is significantly associated with reduced recurrence of NMIBC patients. Importantly, the presence of FGFR3 mutations may influence the clinical outcome of patients bearing alterations in PIK3CA gene, and increased recurrence was associated to FGFR3 mutated, PIK3CA wt tumors. These findings may have high relevance in terms of using PI3K-targeted therapies for BC treatment.

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.

Evaluating the quality of antimicrobial prescribing: is standardisation possible?

The quality of antimicrobial prescribing refers to the optimal way to use antibiotics in regard to their benefits, safety (e.g., resistance generation and toxicity) and cost. Evaluating the quality of antimicrobial prescribing in a way that focuses not only on reducing antimicrobial consumption but also on using them in a more optimal way allows us to understand patterns of use and to identify targets for intervention. The lack of standardisation is the primary problem to be addressed when planning an evaluation of antimicrobial prescribing. There is little information specifically describing an evaluation methodology. Information related to prescription evaluation can be obtained from the guidelines of Antimicrobial Stewardship Programs (ASPs) and from local and international experience. The criteria used to evaluate the quality of prescription should include the indication for antimicrobial therapy, the timeliness of initiation, the correct antibiotic choice (according to local guidelines), the dosing, the duration, the route of administration and the time at which to switch to oral administration. A locally developed guideline on antimicrobial therapy should preferably be the gold standard by which to evaluate the appropriatenes of prescriptions. Various approaches used to carry out the evaluations have been described in the literature. Repeated point-prevalence surveys (PPS) have been proven to be effective in identifying targets for quality improvement. Continuous prospective monitoring allows the identification of more precise intervention points at different times during prescription. The design of the study chosen to perform the evaluation should be adapted according to the resources available in each centre. Evaluating the quality of antimicrobial prescribing should be the first step to designing ASPs, as well as to evaluating their impact and the changes in prescribing trends over time.

The zeta potential of extended dielectrics and conductors in terms of streaming potential and streaming current measurements.

The electrical characterization of surfaces in terms of the zeta potential (ζ), i.e., the electric potential contributing to the interaction potential energy, is of major importance in a wide variety of industrial, environmental and biomedical applications in which the integration of any material with the surrounding media is initially mediated by the physico-chemical properties of its outer surface layer. Among the different existing electrokinetic techniques for obtaining ζ, streaming potential (V(str)) and streaming current (I(str)) are important when dealing with flat-extended samples. Mostly dielectric materials have been subjected to this type of analysis and only a few papers can be found in the literature regarding the electrokinetic characterization of conducting materials. Nevertheless, a standardized procedure is typically followed to calculate ζ from the measured data and, importantly, it is shown in this paper that such a procedure leads to incorrect zeta potential values when conductors are investigated. In any case, assessment of a reliable numerical value of ζ requires careful consideration of the origin of the input data and the characteristics of the experimental setup. In particular, it is shown that the cell resistance (R) typically obtained through a.c. signals (R(a.c.)), and needed for the calculations of ζ, always underestimates the zeta potential values obtained from streaming potential measurements. The consideration of R(EK), derived from the V(str)/I(str) ratio, leads to reliable values of ζ when dielectrics are investigated. For metals, the contribution of conductivity of the sample to the cell resistance provokes an underestimation of R(EK), which leads to unrealistic values of ζ. For the electrical characterization of conducting samples I(str) measurements constitute a better choice. In general, the findings gathered in this manuscript establish a measurement protocol for obtaining reliable zeta potentials of dielectrics and conductors based on the intrinsic electrokinetic behavior of both types of samples.

Hamamelitannin from witch hazel (Hamamelis virginiana) displays specific cytotoxic activity against colon cancer cells.

Hamamelis virginiana (witch hazel) bark is a rich source of condensed and hydrolyzable tannins reported to exert a protective action against colon cancer. The present study characterizes different witch hazel tannins as selective cytotoxic agents against colon cancer. To cover the structural diversity of the tannins that occur in H. virginiana bark, the hydrolyzable tannins, hamamelitannin and pentagalloylglucose, together with a proanthocyanidin-rich fraction (F800H4) were selected for the study. Treatment with these compounds reduced tumor viability and induced apoptosis, necrosis, and S-phase arrest in the cell cycle of HT29 cells, with hamamelitannin being the most efficient. Owing to polyphenol-mediated H(2)O(2) formation in the incubation media, the antiproliferative effect was determined in the presence and absence of catalase to rule out any such interference. The presence of catalase significantly changed the IC(50) only for F800H4. Furthermore, at concentrations that inhibit the growth of HT29 cells by 50%, hamamelitannin had no harmful effects on NCM460 normal colonocytes, whereas pentagalloylglucose inhibited both cancerous and normal cell growth. Using the TNPTM assay, we identified a highly reactive phenolic position in hamamelitannin, which may explain its efficacy at inhibiting colon cancer growth.

Modification of physico-chemical surface properties and growth of Staphylococcus aureus under hyperglycemia and ketoacidosis conditions.

Diabetes is a widely spread disease affecting the quality of life of millions of people around the world and is associated to a higher risk of developing infections in different parts of the body. The reasons why diabetes enhances infection episodes are not entirely clear; in this study our aim was to explore the changes that one of the most frequently pathogenic bacteria undergoes when exposed to hyperglycemia and ketoacidosis conditions. Physical surface properties such as hydrophobicity and surface electrical charge are related to bacterial growth behavior and the ability of Staphylococcus aureus to form biofilms. The addition of glucose made bacteria more negatively charged and with moderate-intermediate hydrophobicity. Ketone bodies increased hydrophobicity to approximately 75% and pathological concentrations hindered some of the bacterial surface charge by decreasing the negative zeta potential of cells. When both components were present, the bacterial physical surface changes were more similar to those observed in ketone bodies, suggesting a preferential adsorption of ketone bodies over glucose because of the more favorable solubility of glucose in water. Glucose diabetic concentrations gave the highest number of bacteria in the stationary phase of growth and provoked an increase in the biofilm slime index of around 400% in relation to the control state. Also, this situation is related with an increase of bacterial coverage. The combination of a high concentration of glucose and ketone bodies, which corresponds to a poorly controlled diabetic situation, appears associated with an early infection phase; increased hydrophobic attractive force and reduced electrostatic repulsion between cells results in better packing of cells within the biofilm and more efficient retention to the host surface. Knowledge of bacterial response in high amount of glucose and ketoacidosis environments can serve as a basis for designing strategies to prevent bacterial adhesion, biofilm formation and, consequently, the development of infections.

The role of magnesium in biomaterials related infections.

Magnesium is currently increasing interest in the field of biomaterials. An extensive bibliography on this material in the last two decades arises from its potential for the development of biodegradable implants. In addition, many researches, motivated by this progress, have analyzed the performance of magnesium in both in vitro and in vivo assays with gram-positive and gram-negative bacteria in a very broad range of conditions. This review explores the extensive literature in recent years on magnesium in biomaterials-related infections, and discusses the mechanisms of the Mg action on bacteria, as well as the competition of Mg2+ and/or synergy with other divalent cations in this subject.

Impact of PLA/Mg films degradation on surface physical properties and biofilm survival.

New biocompatible and bioabsorbable materials are currently being developed for bone regeneration. These serve as scaffolding for controlled drug release and prevent bacterial infections. Films of polylactic acid (PLA) polymers that are Mg-reinforced have demonstrated they have suitable properties and bioactive behavior for promoting the osseointegration process. However little attention has been paid to studying whether the degradation process can alter the adhesive physical properties of the biodegradable film and whether this can modify the biofilm formation capacity of pathogens. Moreover, considering that the concentration of Mg and other corrosion products may not be constant during the degradation process, the question that arises is whether these changes can have negative consequences in terms of the bacterial colonization of surfaces. Bacteria are able to react differently to the same compound, depending on its concentration in the medium and can even become stronger when threatened. In this context, physical surface parameters such as hydrophobicity, surface tension and zeta potential of PLA films reinforced with 10% Mg have been determined before and after degradation, as well as the biofilm formation capacity of Staphylococcus epidermidis. The addition of Mg to the films makes them less hydrophobic and the degradation also reduces the hydrophobicity and increases the negative charge of the surface, especially over long periods of time. Early biofilm formation at 8 h is consistent with the physical properties of the films, where we can observe a reduction in the bacterial biofilm formation. However, after 24 h of incubation, the biofilm formation increases significantly on the PLA/Mg films with respect to PLA control. The explosive release of Mg ions and other corrosion products within the first hours were not enough to prevent a greater biofilm formation after this initial time. Consequently, the Mg addition to the polymer matrix had a bacteriostatic effect but not a bactericidal one. Future works should aim to optimize the design and biofunctionality of these promising bioabsorbable composites for a degradation period suitable for the intended application.

Relevance of Topographic Parameters on the Adhesion and Proliferation of Human Gingival Fibroblasts and Oral Bacterial Strains.

Dental implantology allows replacement of failing teeth providing the patient with a general improvement of health. Unfortunately not all reconstructions succeed, as a consequence of the development of infections of bacterial origin on the implant surface. Surface topography is known to modulate a differential response to bacterial and mammalian cells but topographical measurements are often limited to vertical parameters. In this work we have extended the topographical measurements also to lateral and hybrid parameters of the five most representative implant and prosthetic component surfaces and correlated the results with bacterial and mammalian cell adhesion and proliferation outcomes. Primary human oral gingival fibroblast (gum cells) and the bacterial strains: Streptococcus mutans, Streptococcus sanguinis and Aggregatibacter actinomycetemcomitans, implicated in infectious processes in the oral/implant environment were employed in the presence or absence of human saliva. The results confirm that even though not all the measured surface is available for bacteria to adhere, the overall race for the surface between cells and bacteria is more favourable to the smoother surfaces (nitrided, as machined or lightly acid etched) than to the rougher ones (strong acid etched or sandblasted/acid etched).

Kinetic of Adhesion of S. epidermidis with Different EPS Production on Ti6Al4V Surfaces.

Controlling initial bacterial adhesion is essential to prevent biofilm formation and implant-related infection. The search for surface coatings that prevent initial adhesion is a powerful strategy to obtain implants that are more resistant to infection. Tracking the progression of adhesion on surfaces from the beginning of the interaction between bacteria and the surface provides a deeper understanding of the initial adhesion behavior. To this purpose, we have studied the progression over time of bacterial adhesion from a laminar flow of a bacterial suspension, using a modified Robbins device (MRD). Comparing with other laminar flow devices, such as the parallel plate flow chamber, MRD allows the use of diverse substrata under the same controlled flow conditions simultaneously. Two different surfaces of Ti6Al4V and two strains of Staphylococcus epidermidis with different exopolymer production were tested. In addition, the modified Robbins device was examined for its convenience and suitability for the purpose of this study. Results were analyzed according to a pseudofirst order kinetic. The values of the parameters obtained from this model make it possible to discriminate the adhesive behavior of surfaces and bacteria. One of the fitting parameters depends on the bacterial strain and the other only on the surface properties of the substrate.

Per-6-O-(tert-butyldimethylsilyl)cyclodextrins (TBDMS-CDs) in Langmuir monolayers: the importance of a spreading solvent in the preparation of LB layers suitable for sensor application.

Commercially available amphiphilic cyclodextrins, namely per-6-O-(tert-butyldimethylsilyl) alpha, beta and gamma cyclodextrins (TBDMS-alpha-, -beta-, and -gamma-CDs) were subjected to a thorough Langmuir monolayer characterization, using both traditional methods of surface manometry (pi/A isotherms, stability experiments) and modern micrometer/nanometer resolution (BAM, AFM) surface techniques. It has been found that inconsistent behavior regarding the isotherms reproducibility obtained upon compression of TBDMS-beta-CDs is due to the aggregation of the investigated molecules in chloroform and hexane, while good reproducibility ensured a mixed spreading solvent system of hexane/isopropanol 7:3 (v/v). Although the stability of films dropped from chloroform and hexane/isopropanol solvents below the equilibrium surface pressure (ESP) was comparable, pronounced differences were observed at pressures above ESP. The investigated TBDMS-CDs were successfully transferred onto cadmium stearate covered mica substrates. AFM images confirmed the presence of discontinuous multilayered films (10 nm heights) spread from chloroform versus monomolecular dispersion achieved in hexane/isopropanol.

Los puntos oscuros de la insuficiencia venosa pélvica / The dark spots of pelvic venous insufficiency

La insuficiencia venosa pélvica es un tema de mucho debate en congresos de nuestro ámbito profesional. Sin embargo, cuanto más se reconoce y se trata esta patología en nuestro medio, más casos encontramos en los que, aplicando un tratamiento estándar de embolización venosa o stenting, se obtiene un resultado desfavorable. Para evitar este patrón de resultados, debemos estudiar con detenimiento la circulación venosa abdominopélvica estableciendo paralelismos con la circulación venosa infrainguinal y, además, aplicar tecnología IVUS en los casos de duda. Con estas dos medidas, podemos afrontar esta patología en su conjunto con unos resultados muy positivos para nuestros pacientes

Pelvic Venous Insufficiency is a topic of much debate in Congresses of our professional field. However, the more this pathology is recognized and treated in our setting, the more cases we find in which applying a standard venous embolization or stenting, treatment results in an unfavorable result. To avoid this type of results, we must carefully study the abdomino-pelvic venous circulation, establishing parallels with the infrainguinal venous circulation, and also apply IVUS technology in cases of doubt. With these two measures, we can face this pathology as a whole with very positive results for our patients

Neo-clerodane diterpenoids from Croton schiedeanus.

Two new neo-clerodane type furano diterpenoids were isolated from the aerial part of Croton schiedeanus, besides the clerodane diterpenes cis- and trans-dehydrocrotonin, previously isolated from other species of Croton. Structural elucidation was achieved on basis of extensive NMR experiments, including X-ray diffraction analysis and molecular mechanics calculations. The previously known flavonoids ayanin and quercetin-3,7-dimethyl ether were also obtained from the extract of this plant.

Adsorption behavior of human plasma fibronectin on hydrophobic and hydrophilic Ti6Al4V substrata and its influence on bacterial adhesion and detachment.

Biomaterial implant-associated infections, a common cause of medical devices' failure, are initiated by bacterial adhesion to an adsorbed protein layer on the implant material surface. In this study, the influence of protein surface orientation on bacterial adhesion has been examined using three clinically relevant bacterial strains known to express specific binding sites for human plasma fibronectin (HFN). HFN was allowed to adsorb on hydrophobic Ti6Al4V and physically modified hydrophilic Ti6Al4V substrata. Ellipsometric data reveal that the characteristics of the adsorbed protein layers primary depend on solid surface tension and the initial protein concentration in solution. In particular, HFN molecules adopt a more extended conformation on hydrophobic than hydrophilic surfaces, an effect that is more pronounced at low than at high initial protein concentrations. Moreover, the extended conformation of the protein molecules on these surfaces likely facilitates the exposure of specific sites for adhesion, resulting in the higher bacterial-cell attachment observed regardless of the strain considered. Contact angle measurements and the analysis of the number of remaining adhering cells after being subjected to external forces further suggest that both specific and nonspecific (hydrophobic) interactions play an important role on bacterial attachment. This study is the first one to evaluate the influence of surface hydrophobicity on protein adsorption and its subsequent effect on bacterial adhesion using a material whose hydrophobicity was not modified using chemical treatments that potentially led to surface properties changes other than hydrophobicity.

Electrochemical analysis of the UV treated bactericidal Ti6Al4V surfaces.

This research investigates in detail the bactericidal effect exhibited by the surface of the biomaterial Ti6Al4V after being subjected to UV-C light. It has been recently hypothesized that small surface currents, occurring as a consequence of the electron-hole pair recombination taking place after the excitation process, are behind the bactericidal properties displayed by this UV-treated material. To corroborate this hypothesis we have used different electrochemical techniques, such as electrochemical impedance spectroscopy (EIS), potentiodynamic polarization plots and Mott-Schottky plots. EIS and Mott-Schottky plots have shown that UV-C treatment causes an initial increase on the surface electrical conduction of this material. In addition, EIS and polarization plots demonstrated that higher corrosion currents occur at the UV treated than at the non-irradiated samples. Despite this increase in the corrosion currents, EIS has also shown that such currents are not likely to affect the good stability of this material oxide film since the irradiated samples completely recovered the control values after being stored in dark conditions for a period not longer than 24h. These results agree with the already-published in vitro transitory behavior of the bactericidal effect, which was shown to be present at initial times after the biomaterial implantation, a crucial moment to avoid a large number of biomaterial associated infections.

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.