Solid Sampling with a Diode Laser for Portable Ambient Mass Spectrometry

A hand-held diode laser is implemented for solid sampling in portable ambient mass spectrometry (MS). Specifically, a pseudocontinuous wave battery powered surgical laser diode is employed for portable laser diode thermal desorption (LDTD) at 940 nm and compared with nanosecond pulsed laser ablation at 2940 nm. Postionization is achieved in both cases using atmospheric pressure photoionization (APPI). The laser ablation atmospheric pressure photoionization (LAAPPI) and LDTD-APPI mass spectra of sage leaves (Salvia officinalis) using a field-deployable quadrupole ion trap MS display many similar ion peaks, as do the mass spectra of membrane grown biofilms of Pseudomonas aeruginosa: These results indicate that LDTD-APPI method should be useful for in-field sampling of plant and microbial communities, for example, by portable ambient MS. The feasibility of many portable MS applications is facilitated by the availability of relatively low cost, portable, battery-powered diode lasers. LDTD could also be coupled with plasma- or electrospray-based ionization for the analysis of a variety of solid-samples.Peer reviewe

\u3cem\u3ePseudomonas aeruginosa\u3c/em\u3e reverse diauxie is a multidimensionl, optimized, resource utilization strategy

Pseudomonas aeruginosa is a globally-distributed bacterium often found in medical infections. The opportunistic pathogen uses a different, carbon catabolite repression (CCR) strategy than many, model microorganisms. It does not utilize a classic diauxie phenotype, nor does it follow common systems biology assumptions including preferential consumption of glucose with an ‘overflow’ metabolism. Despite these contradictions, P. aeruginosa is competitive in many, disparate environments underscoring knowledge gaps in microbial ecology and systems biology. Physiological, omics, and in silico analyses were used to quantify the P. aeruginosa CCR strategy known as ‘reverse diauxie’. An ecological basis of reverse diauxie was identified using a genome-scale, metabolic model interrogated with in vitro omics data. Reverse diauxie preference for lower energy, nonfermentable carbon sources, such as acetate or succinate over glucose, was predicted using a multidimensional strategy which minimized resource investment into central metabolism while completely oxidizing substrates. Application of a common, in silico optimization criterion, which maximizes growth rate, did not predict the reverse diauxie phenotypes. This study quantifies P. aeruginosa metabolic strategies foundational to its wide distribution and virulence including its potentially, mutualistic interactions with microorganisms found commonly in the environment and in medical infections

Classical N=2 W-superalgebras From Superpseudodifferential Operators

We study the supersymmetric Gelfand-Dickey algebras associated with the superpseudodifferential operators of positive as well as negative leading order. We show that, upon the usual constraint, these algebras contain the N=2 super Virasoro algebra as a subalgebra as long as the leading order is odd. The decompositions of the coefficient functions into N=1 primary fields are then obtained by covariantizing the superpseudodifferential operators. We discuss the problem of identifying N=2 supermultiplets and work out a couple of supermultiplets by explicit computations.Comment: 19 pages (Plain TeX), NHCU-HEP-94-1

Metabolites and Proteins of Pseudomonas aeruginosa Biofilms

Bacterial biofilms are structurally and physiologically heterogenous communities of bacterial cells embedded in a self-produced extracellular polymeric substance. Current studies aim to understand the dynamics and responses of biofilms to various environmental factors especially since they play a pathogenic role in chronic wounds preventing wound healing. One approach to studying biofilms is through mass spectrometry (MS), a versatile analytical tool that provides qualitative and quantitative analyses and has been utilized for various biological applications such as biofilms. This dissertation describes the development of two mass spectrometric strategies for the detection of metabolites and proteins from a biofilm-forming Pseudomonas aeruginosa chronic wound clinical isolate. Two laser-based ion sources were implemented: atmospheric pressure matrix assisted laser desorption ionization MS and laser diode thermal desorption atmospheric pressure photoionization MS. The latter was demonstrated for the analysis of metabolites within intact P. aeruginosa biofilms. A bottom-up proteomic workflow for planktonic and biofilm cultures was also developed based upon liquid chromatography tandem mass spectrometry. This proteomic protocol was then used to compare the effect of lactate nutrient supplementation on cellular metabolism in both P. aeruginosa planktonic and biofilm cultures

Pseudomonas aeruginosa reverse diauxie is a multidimensional, optimized, resource utilization strategy

Abstract Pseudomonas aeruginosa is a globally-distributed bacterium often found in medical infections. The opportunistic pathogen uses a different, carbon catabolite repression (CCR) strategy than many, model microorganisms. It does not utilize a classic diauxie phenotype, nor does it follow common systems biology assumptions including preferential consumption of glucose with an ‘overflow’ metabolism. Despite these contradictions, P. aeruginosa is competitive in many, disparate environments underscoring knowledge gaps in microbial ecology and systems biology. Physiological, omics, and in silico analyses were used to quantify the P. aeruginosa CCR strategy known as ‘reverse diauxie’. An ecological basis of reverse diauxie was identified using a genome-scale, metabolic model interrogated with in vitro omics data. Reverse diauxie preference for lower energy, nonfermentable carbon sources, such as acetate or succinate over glucose, was predicted using a multidimensional strategy which minimized resource investment into central metabolism while completely oxidizing substrates. Application of a common, in silico optimization criterion, which maximizes growth rate, did not predict the reverse diauxie phenotypes. This study quantifies P. aeruginosa metabolic strategies foundational to its wide distribution and virulence including its potentially, mutualistic interactions with microorganisms found commonly in the environment and in medical infections

Spatially resolved analysis of Pseudomonas aeruginosa biofilm proteomes measured by laser ablation sample transfer

Heterogeneity in the distribution of nutrients and oxygen gradients during biofilm growth gives rise to changes in phenotype. There has been long term interest in identifying spatial differences during biofilm development including clues that identify chemical heterogeneity. Laser ablation sample transfer (LAST) allows site-specific sampling combined with label free proteomics to distinguish radially and axially resolved proteomes for Pseudomonas aeruginosa biofilms. Specifically, differential protein abundances on oxic vs. anoxic regions of a biofilm were observed by combining LAST with bottom up proteomics. This study reveals a more active metabolism in the anoxic region of the biofilm with respect to the oxic region for this clinical strain of P. aeruginosa, despite this organism being considered an aerobe by nature. Protein abundance data related to cellular acclimations to chemical gradients include identification of glucose catabolizing proteins, high abundance of proteins from arginine and polyamine metabolism, and proteins that could also support virulence and environmental stress mediation in the anoxic region. Finally, the LAST methodology requires only a few mm2 of biofilm area to identify hundreds of proteins