Mucus layers often provide a unique and multi-functional hydrogel interface between the epithelial cells of organisms and their external environment. Mucus has exceptional properties including elasticity, changeable rheology and an ability to self-repair by re-annealing, and is therefore an ideal medium for trapping and immobilising pathogens and serving as a barrier to microbial infection. The ability to produce a functional surface mucosa was an important evolutionary step, which evolved first in the Cnidaria, which includes corals, and the Ctenophora. This allowed the exclusion of non-commensal microbes and the subsequent development of the mucus-lined digestive cavity seen in higher metazoans. The fundamental architecture of the constituent glycoprotein mucins is also evolutionarily conserved. Although an understanding of the biochemical interactions between bacteria and the mucus layer are important to the goal of developing new antimicrobial strategies, they remain relatively poorly understood. This review summarises the physicochemical properties and evolutionary importance of mucus, which make it so successful in the prevention of bacterial infection. In addition, the strategies developed by bacteria to counteract the mucus layer are also explored.
The mechanical properties of Rhodococcus RC291 were measured using force spectroscopy equipped with a bacterial cell probe. Rhodococcal cells in the late growth stage of development were found to have greater adhesion to a silicon oxide surface than those in the early growth stage. This is because there are more extracellular polymeric substances (EPS) that contain nonspecific binding sites available on the cells of late growth stage. It is found that EPS in the late exponential phase are less densely bound but consist of chains able to extend further into their local environment, while the denser EPS at the late stationary phase act more to sheath the cell. Contraction and extension of the EPS could change the density of the binding sites, and therefore affect the magnitude of the adhesion force between the EPS and the silicon oxide surface. By treating rhodococcal EPS as a surface-grafted polyelectrolyte layer and using scaling theory, the interaction between EPS and a solid substrate was modelled for the cell approaching the surface which revealed that EPS possess a large capacity to store charge. Changing the pH of the surrounding medium acts to change the conformation of EPS chains.
Force spectroscopy was used to show that extracellular DNA (eDNA) has the pre-eminent structural role in a biofilm. The adhesive behavior of extracellular polymeric substances to poly(ethylene terephthalate), a model hydrophobic surface, was measured in response to their degradation by hydrolytic enzymes known for their biofilm-dispersion potential: DNaseI, protease, cellulase, and mannanase. Only treatment with DNaseI significantly decreased the adhesive force of the model bacterium Micrococcus luteus with the surface, and furthermore this treatment almost completely eliminated any components of the biofilm maintaining the adhesion, establishing a key structural role for eDNA. ASSOCIATED CONTENT Supporting Information. Contact angle methods, results of biofilm coated silicon wafer after enzyme treatments, fluorescence image of biofilm-coated cantilever, and forcedistance control curves showing poly-lysine contacting a PET substrate. This material is available free of charge via the Internet at http://pubs.acs.org.
The inclusion of a nuclease, a new‐to‐laundry enzyme class in detergent formulations, reduces the concentration of recalcitrant soils on dirty clothing. Nucleases target extracellular DNA that, albeit present in minor quantities in soiled laundry items, disproportionately contributes to the accumulation of sebaceous soils and other compounds that affect the appearance of clothes. These lingering soils accumulate over multiple wear cycles and provoke a dingy appearance in clothing, characterized by dull colors, yellow staining, and an unpleasant malodor. The single and continued use of a nuclease in detergent formulations offers increased removal of these recalcitrant soils to produce less malodorous and visually preferred textiles. Furthermore, the nuclease is effective across different detergent forms, as well as in cotton short washing cycles, enabling improved cleaning in more sustainable laundry washing conditions.
Attachment assays of a Pseudomonas isolate to fused silica slides showed that treatment with DNaseI significantly inhibited cellular adsorption, which was restored upon DNA treatment. These assays confirmed the important role of extracellular DNA (eDNA) adsorption to a surface. To investigate the eDNA adsorption mechanism, single-molecule force spectroscopy (SMFS) was used to measure the adsorption of eDNA to silicon surfaces in the presence of different concentrations of sodium and calcium ions. SMFS reveals that the work of adhesion required to remove calcium-bound eDNA from the silicon oxide surface is substantially greater than that for sodium. Molecular dynamics simulations were also performed, and here, it was shown that the energy gain in eDNA adsorption to a silicon oxide surface in the presence of calcium ions is small and much less than that in the presence of sodium. The simulations show that the length scales involved in eDNA adsorption are less in the presence of sodium ions than those in the presence of calcium. In the presence of calcium, eDNA is pushed above the surface cations, whereas in the presence of sodium ions, short-range interactions with the surface dominate. Moreover, SMFS data show that increasing [Ca 2+ ] from 1 to 10 mM increases the adsorption of the cations to the silicon oxide surface and consequently enhances the Stern layer, which in turn increases the length scale associated with eDNA adsorption.
Direct glycan-glycan interactions are increasingly implicated in survival and pathogenicity of bacteria. Here, we show that they can be exploited by protozoan parasites in their insect hosts. Force spectroscopy revealed...
This paper introduces a methodological framework for assessing the sustainability of solid biofuels in Mexico. The designed framework comprises 13 normalized indicators and two diagnostic studies, covering the economic, social, environmental, and institutional sustainability dimensions, and their intersections. Indicators are normalized using the concept of load capacity of a system, similarly to the planetary boundaries. Thus, the graphical representation of results facilitates their multidimensional analysis. The framework was applied to three case studies: traditional fuelwood in rural households, charcoal for restaurant grilling, and electricity cogeneration from sugarcane bagasse. This was part of an iterative process of testing and refining the framework and simultaneously demonstrating its application in the Mexican bioenergy context. This led to the conclusion that the resulting framework (a) provides a useful, quantitative, and comprehensive overview of both broad and specific sustainability aspects of the assessed system; (b) requires a balance of accessible but also scattered or sensitive data, similarly to most existing frameworks; (c) is highly flexible and applicable to both modern and traditional solid biofuels; and (d) is simple to communicate and interpret for a wide audience. Key directions for improvement of the framework are also discussed.
Washed textiles can remain malodorous and dingy due to the recalcitrance of soils. Recent work has found that ‘invisible’ soils such as microbial extracellular DNA (eDNA) play a key role in the adhesion of extracellular polymeric substances that form matrixes contributing to these undesirable characteristics. Here we report the application of an immunostaining method to illustrate the cleaning mechanism of a nuclease (DNase I) acting upon eDNA. Extending previous work that established a key role for eDNA in anchoring these soil matrixes, this work provides new insights into the presence and effective removal of eDNA deposited on fabrics using high-resolution in-situ imaging. Using a monoclonal antibody specific to Z-DNA, we showed that when fabrics are washed with DNase I, the incidence of microbial eDNA is reduced. As well as a quantitative reduction in microbial eDNA, the deep cleaning benefits of this enzyme are shown using confocal microscopy and imaging analysis of T-shirt fibers. To the best of our knowledge, this is the first time the use of a molecular probe has been leveraged for fabric and homecare-related R&D to visualize eDNA and evaluate its removal from textiles by a new-to-laundry DNase enzyme. The approaches described in the current work also have scope for re-application to identify further cleaning technology.
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