This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. SUBPROJECT DESCRIPTION During this funding period we have developed the secondary electrospray ionization mass spectrometry (SESI-MS) technology to the point of submitting our first publication (see 'papers under review', #4), submitted and received funding from NASA-EPSCoR and pilot project funding from the Cystic Fibrosis Foundation (via Dartmouth College) to support related work, and had two papers accepted for publication. It was wisely suggested that I focus the research goals within the group so as to develop a distinct research niche. Therefore, we have focuses ourselves in two areas: P. aeruginosa (Pa) metabolic state determination using SESI-MS and to build on our inositol phosphate (IP) track record, we also focus on Pa utilization of Fe via IP ligands in the lung environment. Project area 1: Pa Mucoidy 85% of Pa clinical isolates from patients with cystic fibrosis exhibit the mucoid phenotype as a consequence of mutations in mucA. Therefore, the first project focuses on the rapid determination of the mucoidy phenotype for Pa as it relates to mutation or degradation of mucA/MucA in the: shake flask, biofilm, and mouse lung environment. We target pathways that link amino acid metabolism and the production of side chains that are volatile, such as Tyr and Trp, as well as, the production of HCN. Project area 2: Pa utilization of Fe via inositol phosphates The second project focuses on the determination of the receptor, import, and sequence of enzymatic steps that lead to Pa utilization of host Fe and inositol phosphates. We build here on our understanding of receptor-ligand interactions as well as our expertise in understanding redox chemistry and the biology of phosphatase production in prokaryotes. Grant submission plans Two grants in the area of inositol phosphate chemistry and transformation via microorganisms have been submitted in the past six months. Three grant targets are on the horizon: NSF CAREER, USDA-NIFA, and an NIH RO1 target of February 2011. The NSF and USDA grants will focus on SESI-MS application to rapid detection of E. coli O157:H7 on ground beef, in an effort to support the SESI-MS pathogen volatile database-building enterprise and niche. The RO1 proposal will likely focus on the metabolic signature of the Pa non-mucoid-mucoid transition in the ENaC mouse model. Publications 2009/10 Accepted/Published 1.Low S.Y., Hill J.E., and Peccia P. 2009 A DNA Aptamer Recognizes the Asp f 1 Allergen of Aspergillus fumigatus Biochemical and Biophysical Research Communications 386 (3) 544-548 2.Johnson, N.R. and Hill, J.E. Phosphorus composition of a poultry manure-amended soil via enzymatic hydrolysis: demonstration of a high-throughput method and hints on enzyme-labile P SSSAJ Publications 2009/10 under review 1.Giles, C.D., Cade-Menun, B., and Hill, J.E. (under review) Phosphorus mobility and transformation in a poultry manure-amended soil tracked over time 2.Giles, C.D., Cade-Menun, B., and Hill, J.E. (under review) The Inositol Phosphates in Soils and Manures: Abundance, Cycling, and Measurement 3.Haznedaroglu B.Z., Zorlu O., Hill J.E., Walker S.L. (under review) Identifying the role of flagella in the transport of motile and nonmotile Salmonella enterica serovars 4.Zhu J, Bean H., Kuo Y., Hill J.E. Fast detection of headspace volatile organic compounds from bacterial cultures by SESI-MS
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