Broad-spectrum in vitro activity of macrophage infectivity potentiator inhibitors against Gram-negative bacteria and Leishmania major

Background The macrophage infectivity potentiator (Mip) protein, which belongs to the immunophilin superfamily, is a peptidyl-prolyl cis/trans isomerase (PPIase) enzyme. Mip has been shown to be important for virulence in a wide range of pathogenic microorganisms. It has previously been demonstrated that small-molecule compounds designed to target Mip from the Gram-negative bacterium Burkholderia pseudomallei bind at the site of enzymatic activity of the protein, inhibiting the in vitro activity of Mip. Objectives In this study, co-crystallography experiments with recombinant B. pseudomallei Mip (BpMip) protein and Mip inhibitors, biochemical analysis and computational modelling were used to predict the efficacy of lead compounds for broad-spectrum activity against other pathogens. Methods Binding activity of three lead compounds targeting BpMip was verified using surface plasmon resonance spectroscopy. The determination of crystal structures of BpMip in complex with these compounds, together with molecular modelling and in vitro assays, was used to determine whether the compounds have broad-spectrum antimicrobial activity against pathogens. Results Of the three lead small-molecule compounds, two were effective in inhibiting the PPIase activity of Mip proteins from Neisseria meningitidis, Klebsiella pneumoniae and Leishmania major. The compounds also reduced the intracellular burden of these pathogens using in vitro cell infection assays. Conclusions These results indicate that Mip is a novel antivirulence target that can be inhibited using small-molecule compounds that prove to be promising broad-spectrum drug candidates in vitro. Further optimization of compounds is required for in vivo evaluation and future clinical applications

Disinfection of Burkholderia pseudomallei in potable water

The effect of chlorine, monochloramine and UV disinfection on the water-borne pathogen Burkholderia pseudomallei was assessed. Persistence of B. pseudomallei was verified by MPN involving a one-step recovery procedure. Chlorine proved the most effective disinfectant with a 99.99% reduction of a 106 CFU/mL pure bacterial culture followed by 99.9% reduction by monochloramine and 99% reduction by UV. Co-culture of B. pseudomallei with Acanthamoeba astronyxis was found to greatly enhance survival of B. pseudomallei in the presence of all disinfecting agents tested. For example, when amoebae were present 100 times more monochloramine was required to maintain the disinfectant efficacy. Given the results obtained from these co-culture experiments, more research is needed to investigate the role of amoeba and biofilms in survival of B. pseudomallei in potable water

The effect of free chlorine on Burkholderia pseudomallei in potable water

Chlorine is widely used in public water supplies to provide a disinfection barrier. The effect of chlorine disinfection on the water-borne pathogen Burkholderia pseudomallei was assessed using multiple techniques. After exposure to chlorine viable bacteria were undetectable by conventional plate count techniques; however, persistence of B. pseudomallei was verified by flow cytometry and bacteria were recoverable following a simple one-step broth procedure. The minimum residual chlorine concentration and contact time as prescribed by potable water providers in Australia was insufficient to reduce a B. pseudomallei population by more than 2 log10. Chlorine had a bacteriostatic effect only on B. pseudomallei; viable bacteria were recovered from water containing up to 1000 ppm free chlorine. This finding has practical implications for water treatment in regions where B. pseudomallei is endemic. Future work to assess the effect of alternative water disinfection processes either singly or in sequence is necessary

Challenges in diagnosis and treatment of melioidosis

DOI: http://dx.doi.org/10.4038/sljid.v2i1.4067Sri Lankan Journal of Infectious Diseases Vol.2(1) 2012: 52-53</p

Teaching problem-solving skills by analogy

Medical Teacher18173-74MEDT

New insights into the pathogenesis of ventilator-associated pneumonia

Journal of Hospital Infection30SUPPL.409-413JHIN

Rapid genotypic confirmation of methicillin resistance

10.1080/00313029600169114Pathology283259-261PTLG

Dissemination of respiratory secretions during tracheal tube suctioning in an intensive care unit

Annals of the Academy of Medicine Singapore282178-182AAMS

Teaching microbiology with hypertext: First steps towards a virtual textbook

Medical Education296393-396MEDU