The impact of low-shear stress (LSS) was evaluated on an Adherent-invasive Escherichia coli clinical isolate (AIEC strain O83:H1) from a Crohn's disease patient. High-aspect ratio vessels (HARVs) were used to model LSS conditions to characterize changes in environmental stress resistance and adhesion/invasive properties. Low-shear stress-grown cultures exhibited enhanced thermal and oxidative stress resistance as well as increased adherence to Caco-2 cells, but no changes in invasion were observed. An AIEC rpoS mutant was constructed to examine the impact of this global stress regulator. The absence of RpoS under LSS conditions resulted in increased sensitivity to oxidative stress while adherence levels were elevated in comparison with the wild-type strain. TnphoA mutagenesis and rpoS complementation were carried out on the rpoS mutant to identify those factors involved in the LSS-induced adherence phenotype. Mutagenesis results revealed that one insertion disrupted the tnaB gene (encoding tryptophan permease) and the rpoS tnaB double mutant exhibited decreased adherence under LSS. Complementation of the tnaB gene, or medium supplemented with exogenous indole, restored adhesion of the rpoS tnaB mutant under LSS conditions. Overall, our study demonstrated how mechanical stresses such as LSS altered AIEC phenotypic characteristics and identified novel functions for some RpoS-regulated proteins.
We characterize the operating modes of a magnetized cold cathode plasma in helium at pressures 50-6400 mTorr (6.7-850 Pa). We find at least three distinct modes: (i) a diffuse magnetron plasma with representative plasma voltage 250 V; (ii) a constricted mode with a bright spot at the cathode surface that moves slowly, with representative plasma voltage 50 V; and, (iii) a constricted mode (CO) with a bright spot at the cathode surface that orbits rapidly around the racetrack region defined by the magnetic field, with representative plasma voltage 80 V. This latter mode is of interest because it appears to be a cold-cathode plasma operating at unusually low voltage, and it is associated with a low rate of cathode massloss. We find that the CO mode appears only if the initial rate of current-rise is high, and if the cathode surface is smooth and featureless. It also appears to be correlated with cathode materials that form strong oxide films that can dramatically increase the field emission of electrons from the cathode surface.
The gastrointestinal tract is a biologically diverse and complicated system which carries out essential physiological functions that support human health, while at the same time maintaining itself as an isolated environment to prevent infection and systemic disease. To maintain homeostasis in the gut, communication between the host and residing microbial communities must occur to identify and eliminate potential pathogens which could colonize and cause damage through aggressive pro-inflammatory responses by the mucosal immune system. To prevent such events, a number of host and bacterial-mediated mechanisms are utilized to monitor the environment and initiate appropriate immune responses to invading pathogens. An essential component of this communication process between gastrointestinal microflora and the host involves distinguishing indigenous species from pathogens through ligand-receptor interactions which lead to various signaling events in host cells. Such events generally result in the development of mucosal immunity and immunological tolerance. While these signaling pathways provide a highly effective means of communication between the gut microflora and the host, pathogens have developed mechanisms to manipulate these pathways to evade detection by the immune system to persist and cause disease. These adaptations include cell surface modifications and the expression of various virulence factors in response to different immunological and hormonal components produced by the host.
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