Colistin is an antimicrobial peptide that has become the only remaining alternative for the treatment of multidrug-resistant Gram-negative bacterial infections, but little is known of how clinical levels of colistin resistance evolve. We use in vitro experimental evolution and whole-genome sequencing of colistin-resistant Pseudomonas aeruginosa isolates from cystic fibrosis patients to reconstruct the molecular evolutionary pathways open for high-level colistin resistance. We show that the evolution of resistance is a complex, multistep process that requires mutation in at least five independent loci that synergistically create the phenotype. Strong intergenic epistasis limits the number of possible evolutionary pathways to resistance. Mutations in transcriptional regulators are essential for resistance evolution and function as nodes that potentiate further evolution towards higher resistance by functionalizing and increasing the effect of the other mutations. These results add to our understanding of clinical antimicrobial peptide resistance and the prediction of resistance evolution.
Aims: To examine sensitivities of various Drosophila melanogaster strains towards human pathogenic and nonpathogenic gram‐positive bacteria.
Methods and Results: The D. melanogaster Oregon R strain was infected by injecting the thorax with a needle containing Escherichia coli (negative control), Listeria monocytogenes, Staphylococcus aureus (both food‐borne pathogens), Listeria innocua, Bacillus subtilis, Carnobacterium maltaromaticum, Lactobacillus plantarum or Pediococcus acidilactici (all nonpathogenic bacteria). Listeria monocytogenes and S. aureus killed the host rapidly compared with the negative control. Infection with L. innocua, B. subtilis or C. maltaromaticum also resulted in a high fly mortality, whereas Lact. plantarum and P. acidilactici resulted in a slightly increased mortality. Four additional D. melanogaster lines, three of which had been selected for heat, cold and desiccation resistance respectively, were subjected to infection by L. monocytogenes, S. aureus and E. coli. Mortality rates were comparable with that of the Oregon R strain.
Conclusions: Use of the injection method shows the limitation of D. melanogaster as a model host for gram‐positive bacteria as opportunistic infection by nonpathogenic gram‐positive bacteria results in partial or high mortality. In addition, lines of fruit flies resistant to various stress exposures did not show an increased resistance to infection by gram‐positive pathogens under the conditions tested.
Significance and Impact of the Study: This study demonstrates the inadequacy of D. melanogaster infected by the injection method in order to distinguish between virulent and nonvirulent gram‐positive bacteria.
High-quality mass spectrometry data of the oscillatory behavior of CO oxidation on SiO(2) supported Pt-nanoparticles at atmospheric pressure have been acquired as a function of pressure, coverage, gas composition and nanoparticle size. The oscillations are self-sustained for several days at constant temperature, pressure and CO/O(2) ratio. The frequency of the oscillations is very well defined and increases over time. The oscillation frequency is furthermore strongly temperature dependent with increasing temperature resulting in increasing frequency. A plausible mechanism for the oscillations is proposed based on an oxidation-reduction cycle of the nanoparticles which change the rate of CO oxidation on the particles.
We demonstrate a combined microreactor and time of flight system for testing and characterization of heterogeneous catalysts with high resolution mass spectrometry and high sensitivity. Catalyst testing is performed in silicon-based microreactors which have high sensitivity and fast thermal response. Gas analysis is performed with a time of flight mass spectrometer with a modified nude Bayard-Alpert ionization gauge as gas ionization source. The mass resolution of the time of flight mass spectrometer using the ion gauge as ionization source is estimated to m/Δm > 2500. The system design is superior to conventional batch and flow reactors with accompanying product detection by quadrupole mass spectrometry or gas chromatography not only due to the high sensitivity, fast temperature response, high mass resolution, and fast acquisition time of mass spectra but it also allows wide mass range (0–5000 amu in the current configuration). As a demonstration of the system performance we present data from ammonia oxidation on a Pt thin film showing resolved spectra of OH and NH3.
The computer programs described here plot students' titration data, locate equivalence points of the titration, calculate the apparent equivalent mass of the acid, and determine the successive ionization constants of the acid.
In this article, a flexible free and open-source software system for data logging and presentation will be described. The system is highly modular and adaptable and can be used in any laboratory in which continuous and/or ad hoc measurements require centralized storage. A presentation component for the data back end has furthermore been written that enables live visualization of data on any device capable of displaying Web pages. The system consists of three parts: data-logging clients, a data server, and a data presentation Web site. The logging of data from independent clients leads to high resilience to equipment failure, whereas the central storage of data dramatically eases backup and data exchange. The visualization front end allows direct monitoring of acquired data to see live progress of long-duration experiments. This enables the user to alter experimental conditions based on these data and to interfere with the experiment if needed. The data stored consist both of specific measurements and of continuously logged system parameters. The latter is crucial to a variety of automation and surveillance features, and three cases of such features are described: monitoring system health, getting status of long-duration experiments, and implementation of instant alarms in the event of failure.
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