A novel targeted/untargeted GC-Orbitrap metabolomics methodology applied to Candida albicans and Staphylococcus aureus biofilms

Introduction: Combined infections from Candida albicans and Staphylococcus aureus are a leading cause of death in the developed world. Evidence suggests that Candida enhances the virulence of Staphylococcus—hyphae penetrate through tissue barriers, while S. aureus tightly associates with the hyphae to obtain entry to the host organism. Indeed, in a biofilm state, C. albicans enhances the antimicrobial resistance characteristics of S. aureus. The association of these microorganisms is also associated with significantly increased morbidity and mortality. Due to this tight association we hypothesised that metabolic effects were also in evidence. Objectives: To explore the interaction, we used a novel GC-Orbitrap-based mass spectrometer, the Q Exactive GC, which combines the high peak capacity and chromatographic resolution of gas chromatography with the sub-ppm mass accuracy of an Orbitrap system. This allows the capability to leverage the widely available electron ionisation libraries for untargeted applications, along with expanding accurate mass libraries and targeted matches based around authentic standards. Methods: Optimised C. albicans and S. aureus mono- and co-cultured biofilms were analysed using the new instrument in addition to the fresh and spent bacterial growth media. Results: The targeted analysis experiment was based around 36 sugars and sugar phosphates, 22 amino acids and five organic acids. Untargeted analysis resulted in the detection of 465 features from fresh and spent medium and 405 from biofilm samples. Three significantly changing compounds that matched to high scoring library fragment patterns were chosen for validation. Conclusion: Evaluation of the results demonstrates that the Q Exactive GC is suitable for metabolomics analysis using a targeted/untargeted methodology. Many of the results were as expected: e.g. rapid consumption of glucose and fructose from the medium regardless of the cell type. Modulation of sugar-phosphate levels also suggest that the pentose phosphate pathway could be enhanced in the cells from co-cultured biofilms. Untargeted metabolomics results suggested significant production of cell-wall biosynthesis components and the consumption of non-proteinaceous amino-acids

Improving aircraft stress-loads interface and evaluation procedures

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

QUASI-REGULAR ORTHOGONAL SYSTEMS OF SUBSPACES

Document: Verbal Argument on Mohave County's need for an exchangeable waters survey under SB 58, by William C. Porter, August 7, 1963, page 1VERBAL ARGUMENT ON MOHAVE COUNTY'S NEED FOR AN EXCHANGEABLE WATERS SURVEY UNDER SB 58 By William C. Porter, Special Water Assistant to Robert Peart Presented by Robert Peart, Mohave County Supervisor, on August 7, 1963 BEFORE THE ARIZONA INTER-STATE STREAM COMMISSION Gentlemen... Our presentation this morning will not be lengthy or time-consuming. We have come to you with a relatively simple request, but a necessary and important one. We wish first to give you some background on our request, then state our request in definite terms and answer any questions you may have. Mohave County and its principal city, Kingman, find themselves in a complex position. The Northwestern part of this state is among the most rapidly growing and expanding in the nation. Yet, for the moment we remain the least populous county in the state. We have more shoreline and more water flowing around our boundaries in Mohave County than almost any other county in the nation. Yet, three weeks ago, a water emergency was declared in the City of Kingman. Water useage in the county today is far above the highest estimates made three years ago for the present period. Yet, there is confusion and lack of definite factual data on waters in the county... where they lie and which of the surface waters are feasible for exchange for Colorado River waters. This is one fact that is known...Mohave County must have an allocation from the Colorado River. We hope there are waters we can exchange, but if surveys should show we have not, we will have to seek a direct, straight-across allocation of a fair share of Arizona's 2,800,000 Acre-feet for Mohave County. To fill in some of the details on these opening statements...here are some facts: -1-Epson Perfection 4870 Photo, 400 dpi, 8 bit, 2,488,687 byte

αμ\alpha\mu-Duals and holomorphic (nuclear) mappings

Corresponding to an arbitrary sequence space $\mu$ and a sequence $\alpha$, we introduce the notion of an $\alpha\mu$-dual of a sequence space which, in particular, envelops the concepts of Köthe, $\beta-, \gamma-$ duals and the duals of an $\mathfrak{g}$-space studied in [4]. Using these concepts, we make a structural study of several subspaces of holomorphic mappings including characterizations of bounded and compact subset