EQUIPMENT-HPLC

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In Project 2, we examine the connection between the bacterial catabolism of host phosphatidylcholine and the induction of bacterial genes involved in host colonization.This pilot project was aimed at purchasing equipment to follow the degradation of phospholipids and to monitor the accumulation of intermediates in the PC catabolic pathway. While we initially pursued the purchase of an HPLC, after multiple interactions with technical experts from Aligent and Shimadzu, it became clear that LC-MS was the more appropriate technique for the sensitive and quantitative studies that would be required by this project and our lab is not equipped to invest the funds or time to establish LC-MS capabilities as this technology is already available to COBRE investigators. In light of this, we successfully piloted a number of colorimetric and fluorescent assays for the detection of these compounds. To enable our lab to use these assays on a routine basis in our laboratory, we have used pilot funds to purchase a microtiter plate reader with the necessary software and capabilities that will enable us to monitor PC catabolism and catabolic intermediates. Our project has also turned to focus on genes involved in bacterial nitrate respiration and their connection to host colonization. The spectrophotometer will also be used to follow nitrogen oxides via colorimetric assays.