Adsorption of aminefluorides on human enamel

Changes in surface characteristics of ground and polished human enamel after adsorption of two types of aminefluorides (AmF 297 and AmF 335) have been studied. After adsorption of aminefluorides from solutions with concentrations up to 10 mM for 2 min followed by rinsing of the surface with distilled water, contact angle measurements were carried out to yield surface free energies and ellipsometry was performed to yield the adsorbed layer thickness. In a separate experiment on powdered enamel, set up in an analogous way, zeta potential changes after adsorption of aminefluorides were determined in a 10 mM potassium phosphate buffer at pH 7·0. Surface free energies decreased from 88 erg·cm−2 to 52 erg·cm−2 and 35 erg·cm−1 after adsorption of AmF 297 and AmF 335 respectively at c = 1 mM. Increasing the aminefluoride concentration in solution did not affect the values obtained. Zeta potentials, originally −36 mV, became positive after adsorption, while ellipsometry indicated the buildup of adsorbed layers with a thickness between 3 run and 12 nm. All three types of experiments indicated that both AmF 297 and AmF 335 form an adsorbed monolayer on ground and polished enamel at a concentration of 1 mM. Negligible additional adsorption takes place at higher concentrations under the present experimental circumstances. In vivo, adsorbed aminefluoride layers will be rapidly covered by adsorbed protein layers, shielding both the adsorbed aminefluoride layer as well as its physicochemical characteristics. This effect has been studied in vivo by measuring surface free energy changes of ground and polished enamel, with AmF 297 and AmF 335 adsorbed at c = 2·5 mM as a function of the time, these samples were carried by test persons in partial dentures. On both types of AmF-coated enamel the surface free energies increased within 30 min to values approaching the one obtained previously for pellicle-coated ground and polished enamel (110 ± 9 erg·cm−2)

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

Factors determining a successful socioeconomic introduction of horticulture in foreign countries - Academic Consultancy Training Report

Course: Academic Consultancy Training (YMC 60809) Project: Sustainable development of greenhouse horticulture in developing countries (756) Commissioner: Wageningen UR Greenhouse Horticulture Contact person: Ir. C.J.M. van der Lans Coach: Dr. Ir. J.W. Hofstee Expert: Prof. Dr. O. van Kooten A lot of capital is attracted in the initiation of Dutch horticulture businesses abroad. There is however a lack of knowledge about what is known about the critical factors determining success or failure of a horticultural initiative in a foreign country. The goal of this project is to achieve socio-economic sustainability of the greenhouse horticulture in foreign countries. Therefore a model is presented which contains all the necessary factors that an entrepreneur should keep in mind to achieve socio-economic sustainability. In order to get an overview of all the important factors literature was studied and interviews were conducted. This led to a detailed description of all the issues that should be considered when setting-up businesses abroad. Those factors are represented in a ‘dial’ model pointing out what issues an entrepreneur should consider. The model is subdivided in three topics: market, production and trade & logistics. Important factors related to the market are whether there is a market and how to influence the whole production chain. Important factors for the production are the physical place and the employees. A very important factor of the trading is the legalization and the bureaucracy of the country you are producing in

Inheritance of physico-chemical properties and ROS generation by carbon quantum dots derived from pyrolytically carbonized bacterial sources

Bacteria are frequently used in industrial processes and nutrient supplementation to restore a healthy human microflora, but use of live bacteria is often troublesome. Here, we hypothesize that bacterially-derived carbon-quantum-dots obtained through pyrolytic carbonization inherit physico-chemical properties from probiotic and pathogenic source-bacteria. Carbon-quantum-dots carbonized at reaction-temperatures below 200 ​°C had negligible quantum-yields, while temperatures above 220 ​°C yielded poor water-suspendability. Fourier-transform infrared-spectroscopy demonstrated preservation of amide absorption bands in carbon-quantum-dots derived at intermediate temperatures. X-ray photoelectron-spectroscopy indicated that the at%N in carbon-quantum-dots increased with increasing amounts of protein in source-bacterial surfaces. Carbonization transformed hydrocarbon-like bacterial surface compounds into heterocyclic aromatic-carbon structures, evidenced by a broad infrared absorption band (920-900 ​cm−1) and the presence of carbon in C–C functionalities of carbon-quantum-dots. The chemical composition of bacterially-derived carbon-quantum-dots could be explained by the degradation temperatures of main bacterial cell surface compounds. All carbon-quantum-dots generated reactive-oxygen-species, most notably those derived from probiotic lactobacilli, carrying a high amount of surface protein. Concluding, amide functionalities in carbon-quantum-dots are inherited from surface proteins of source-bacteria, controlling reactive-oxygen-species generation. This paves the way for applications of bacterially-derived carbon-quantum-dots in which reactive-oxygen-species generation is essential, instead of hard-to-use live bacteria, such as in food supplementation or probiotic-assisted antibiotic therapy

Extracellular Polymeric Matrix Production and Relaxation under Fluid Shear and Mechanical Pressure in Staphylococcus aureus Biofilms

The viscoelasticity of a biofilm's EPS (extracellular polymeric substance) matrix conveys protection against mechanical challenges, but adaptive responses of biofilm inhabitants to produce EPS are not well known. Here, we compare the responses of a biofilm of an EPS-producing (ATCC 12600) and a non-EPS producing (5298) Staphylococcus aureus strain to fluid shear and mechanical challenge. Confocal laser scanning microscopy confirmed absence of calcofluor-white-stainable EPS in biofilms of S. aureus 5298. Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy combined with tribometry indicated that polysaccharide production per bacterium in the initial adhering layer was higher during growth at high shear than at low shear and that this increased EPS production extended to entire biofilms, as indicated by tribometrically measured coefficients of friction (CoF). CoF of biofilms grown under high fluid shear were higher than those when grown under low shear, likely due to wash-off polysaccharides. Measurement of a biofilm's CoF implies application of mechanical pressure that yielded an immediate increase in the polysaccharide band area of S. aureus ATCC 12600 biofilms due to their compression. Compression decreased after relief of pressure to the level observed prior to mechanical pressure. For biofilms grown under high shear, this coincided with a higher percent whiteness in optical coherence tomography-images indicative of water outflow, returning back into the biofilm during stress relaxation. Biofilms grown under low shear, however, were stimulated during tribometry to produce EPS, also after relief of stress. Knowledge of factors that govern EPS production and water flow in biofilms will allow better control of biofilms under mechanical challenge and better understanding of the barrier properties of biofilms against antimicrobial penetration. IMPORTANCE Adaptive responses of biofilm inhabitants in nature to environmental challenges such as fluid shear and mechanical pressure often involve EPS production with the aim of protecting biofilm inhabitants. EPS can assist biofilm bacteria in remaining attached or can impede antimicrobial penetration. The TriboChemist is a recently introduced instrument, allowing the study of initially adhering bacteria to a germanium crystal using ATR-FTIR spectroscopy, while simultaneously allowing measurement of the coefficient of friction of a biofilm, which serves as an indicator of the EPS content of a biofilm. EPS production can be stimulated by both fluid shear during growth and mechanical pressure, while increased EPS production can continue after pressure relaxation of the biofilm. Since EPS is pivotal in the protection of biofilm inhabitants against mechanical and chemical challenges, knowledge of the factors that make biofilm inhabitants decide to produce EPS, as provided in this study, is important for the development of biofilm control measures

Visualization of Bacterial Colonization and Cellular Layers in a Gut-on-a-Chip System Using Optical Coherence Tomography

Imaging of cellular layers in a gut-on-a-chip system has been confined to two-dimensional (2D)-imaging through conventional light microscopy and confocal laser scanning microscopy (CLSM) yielding three-dimensional- and 2D-cross-sectional reconstructions. However, CLSM requires staining and is unsuitable for longitudinal visualization. Here, we compare merits of optical coherence tomography (OCT) with those of CLSM and light microscopy for visualization of intestinal epithelial layers during protection by a probiotic Bifidobacterium breve strain and a simultaneous pathogen challenge by an Escherichia coli strain. OCT cross-sectional images yielded film thicknesses that coincided with end-point thicknesses derived from cross-sectional CLSM images. Light microscopy on histological sections of epithelial layers at the end-point yielded smaller layer thicknesses than OCT and CLSM. Protective effects of B. breve adhering to an epithelial layer against an E. coli challenge included the preservation of layer thickness and membrane surface coverage by epithelial cells. OCT does not require staining or sectioning, making OCT suitable for longitudinal visualization of biological films, but as a drawback, OCT does not allow an epithelial layer to be distinguished from bacterial biofilms adhering to it. Thus, OCT is ideal to longitudinally evaluate epithelial layers under probiotic protection and pathogen challenges, but proper image interpretation requires the application of a second method at the end-point to distinguish bacterial and epithelial films

Surface thermodynamic homeostasis of salivary conditioning films through polar–apolar layering

Salivary conditioning films (SCFs) form on all surfaces exposed to the oral cavity and control diverse oral surface phenomena. Oral chemotherapeutics and dietary components present perturbations to SCFs. Here we determine the surface energetics of SCFs through contact angle measurements with various liquids on SCFs following perturbations with a variety of chemotherapeutics as well as after renewed SCF formation. Sixteen-hour SCFs on polished enamel surfaces were treated with a variety of chemotherapeutics, including toothpastes and mouthrinses. After treatment with chemotherapeutics, a SCF was applied again for 3 h. Contact angles with four different liquids on untreated and treated SCF-coated enamel surfaces were measured and surface free energies were calculated. Perturbations either caused the SCF to become more polar or more apolar, but in all cases, renewed SCF formation compensated these changes. Thus, a polar SCF attracts different salivary proteins or adsorbs proteins in a different conformation to create a more apolar SCF surface after renewed SCF formation and vice versa for apolar SCFs. This polar–apolar layering in SCF formation presents a powerful mechanism in the oral cavity to maintain surface thermodynamic homeostasis—defining oral surface properties within a narrow, biological range and influencing chemotherapeutic strategies. Surface chemical changes brought about by dietary or chemotherapeutic perturbations to SCFs make it more polar or apolar, but new SCFs are rapidly formed compensating for changes in surface energetics

Structural changes in S. epidermidis biofilms after transmission between stainless steel surfaces

Transmission is a main route for bacterial contamination, involving bacterial detachment from a donor and adhesion to receiver surfaces. This work aimed to compare transmission of an extracellular polymeric substance (EPS) producing and a non-EPS producing Staphylococcus epidermidis strain from biofilms on stainless steel. After transmission, donor surfaces remained fully covered with biofilm, indicating transmission through cohesive failure in the biofilm. Counter to the numbers of biofilm bacteria, the donor and receiver biofilm thicknesses did not add up to the pre-transmission donor biofilm thickness, suggesting more compact biofilms after transmission, especially for non-EPS producing staphylococci. Accordingly, staphylococcal density per unit biofilm volume had increased from 0.20 to 0.52 μm(-3) for transmission of the non-EPS producing strain under high contact pressure. The EPS producing strain had similar densities before and after transmission (0.17 μm(-3)). This suggests three phases in biofilm transmission: (1) compression, (2) separation and (3) relaxation of biofilm structure to its pre-transmission density in EPS-rich biofilms

Influence of biofilm lubricity on shear-induced transmission of staphylococcal biofilms from stainless steel to silicone rubber

In real-life situations, bacteria are often transmitted from biofilms growing on donor surfaces to receiver ones. Bacterial transmission is more complex than adhesion, involving bacterial detachment from donor and subsequent adhesion to receiver surfaces. Here, we describe a new device to study shear-induced bacterial transmission from a (stainless steel) pipe to a (silicone rubber) tube and compare transmission of EPS-producing and non-EPS-producing staphylococci. Transmission of an entire biofilm from the donor to the receiver tube did not occur, indicative of cohesive failure in the biofilm rather than of adhesive failure at the donor-biofilm interface. Biofilm was gradually transmitted over an increasing length of receiver tube, occurring mostly to the first 50cm of the receiver tube. Under high-shearing velocity, transmission of non-EPS-producing bacteria to the second half decreased non-linearly, likely due to rapid thinning of the lowly lubricious biofilm. Oppositely, transmission of EPS-producing strains to the second tube half was not affected by higher shearing velocity due to the high lubricity and stress relaxation of the EPS-rich biofilms, ensuring continued contact with the receiver. The non-linear decrease of ongoing bacterial transmission under high-shearing velocity is new and of relevance in for instance, high-speed food slicers and food packaging

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