Further analysis of clinical data and studies involving additiona

Further analysis of clinical data and studies involving additional sets of patients for verification of this hypothesis will provide a clearer picture helping to link genetic features with evidence-led clinical management of P. aeruginosa keratitis. The Microbiology Ophthalmic Group (MOG) includes Stephen Tuft, Stephen Kaye, Timothy Neal, Derek Tole, John Leeming, Peter McDonnell, Francisco Figueiredo, Fiona Carley, Malcolm Armstrong, Colin WIlloughby, Johnny Moore, Grace Ong. This work was supported by the UKNIHR

and a Dr Hans and Mrs Gertrude Hirsch Awards Scheme Fight for Sight Small Projects Grant. S.T. is supported by the NIHR Biomedical Research Centre for Ophthalmology, Moorfields GSK-3 beta pathway Eye Hospital. J.S. and H.S. contributed equally to this work. “
“Nhe (‘nonhaemolytic enterotoxin’) is a three-component cytotoxin implicated in the pathogenesis of diarrhoea Selleckchem Temozolomide by Bacillus

cereus. Nhe forms pores in pure lipid bilayers, but the function of the individual components (NheA,NheB and NheC) remains unclear. NheB and NheC are structural homologues of ClyA, a pore-forming cytotoxin of Escherichia coli. The non-ionic detergent dodecyl maltoside (DDM) has been shown to inhibit haemolysis of ClyA. We used DDM as a probe to examine the response of the Nhe proteins to DDM micelles. At its critical micellar concentration (0.2 mM), DDM inhibited propidium uptake by the native Nhe complex in Vero and HT29 cell suspensions. Pre-incubation of NheC with DDM did not inhibit cytotoxicity. NheB exhibited marked changes in 1-anilinonaphthalene-8-sulphonic acid (ANS) fluorescence after pre-exposure to DDM. Pre-incubation of NheB with DDM resulted in large molecular weight complexes as detected by size exclusion chromatography and diffusion through sized dialysis membranes and prevented binding of NheB to Vero cell monolayers. These data support a model in which conformational changes and oligomerization of NheB are prerequisite events in the 2-hydroxyphytanoyl-CoA lyase process of pore formation. The mechanisms by which Bacillus cereus causes diarrhoea in man remain unknown. Two of the putative enterotoxins, haemolysin BL (HBl) and Nhe (see Stenfors Arnesen et al., 2008), are three-component

cytotoxins. Nhe is cytolytic against both erythrocytes and epithelia because of osmotic lysis induced by pores formed in the host cell plasma membrane (Fagerlund et al., 2008). In epithelial cells, all three Nhe components are necessary for maximal cytotoxic activity (Lund & Granum, 1997). However, in certain cell types, namely rat pituitary GH4 cells, only NheA and NheB are necessary for pore formation and cytotoxicity (Haug et al., 2010). NheB and NheC have significant amino acid sequence homology to the HBl proteins. Using the crystal structure of HBl-B as the template, homology modelling predicts that two of the Nhe components, NheB and NheC, possess marked structural resemblance to ClyA of Escherichia coli (Fagerlund et al., 2008).

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