PSEUDOMONAS AERUGINOSA AND LUNG EPITHELIAL CELL CO-CULTURE METABOLOMICS

铜绿假单胞菌和肺上皮细胞共培养代谢组学

• 批准号: 8167735
• 负责人: Jack Hill
• 金额: $ 16.84万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8167735
• 关键词: Allergens; Amino Acids; Area; Aspergillus fumigatus; Biochemical; Biology; Cade; Chemistry; Clinical; Coculture Techniques; Communication Research; Computer Retrieval of Information on Scientific Projects Database; Cystic Fibrosis; DNA; Databases; Detection; Domestic Fowls; Environment; Enzymes; Epithelial Cells; Escherichia coli O157; Exhibits; Flagella; Foundations; Funding; Goals; Grant; Hydrolysis; Inositol Phosphates; Institution; Lead; Ligands; Link; Lung; Manure; Measurement; Metabolic; Metabolism; Methods; Microbial Biofilms; Mus; Mutation; Oxidation-Reduction; Paper; Pathway interactions; Phenotype; Phosphoric Monoester Hydrolases; Phosphorus; Pilot Projects; Production; Prokaryotic Cells; Pseudomonas aeruginosa; Publications; Publishing; Research; Research Personnel; Resources; Role; Salmonella enterica; Side; Soil; Source; Spectrometry, Mass, Electrospray Ionization; Technology; Time; United States National Institutes of Health; Walkers; Work; aptamer; bean; beef; career; college; cystic fibrosis patients; epithelial Na+ channel; flasks; metabolomics; microorganism; mouse model; mucoid; pathogen; rapid detection; receptor; tryptophyltyrosine; volatile organic compound

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. 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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 子项目描述 在这一资助期间，我们开发了二次电喷雾电离质谱仪(SESI-MS)技术，即将提交我们的第一份出版物(见“正在审查的论文”，#4)，提交并接受了NASA-EPSCoR的资金和囊性纤维化基金会(通过达特茅斯学院)的试点项目资金，以支持相关工作，并接受了两篇论文以供发表。 明智的建议是，我将研究目标集中在小组内，以便开发一个独特的研究利基市场。因此，我们专注于两个领域：使用SESI-MS确定铜绿假单胞菌(PA)的代谢状态；为了建立我们的肌醇磷酸(IP)记录，我们还关注PA通过IP配体在肺环境中对铁的利用。 项目区1：PA Mucoidy 囊性纤维化患者中85%的PA临床分离株表现为粘液样表型，这是粘液A基因突变的结果。因此，第一个项目的重点是快速确定PA的粘液性表型，因为它与摇瓶、生物膜和小鼠肺环境中粘液A/粘液A的突变或降解有关。我们的目标是连接氨基酸代谢和产生挥发性侧链的途径，如Tyr和Trp，以及HCN的产生。 项目领域2：通过肌醇磷酸盐利用铁 第二个项目的重点是确定受体、进口和导致PA利用宿主铁和肌醇磷酸盐的酶步骤序列。我们在这里建立在我们对受体-配体相互作用的理解以及我们在了解氧化还原化学和原核生物中磷酸酶产生的生物学方面的专业知识的基础上。 拨款提交计划 在过去六个月中，提交了两笔关于肌醇磷酸盐化学和微生物转化领域的赠款。有三个拨款目标即将出现：NSF CARAME、USDA-NIFA和NIH 2011年2月的RO1目标。NSF和美国农业部的拨款将集中在SESI-MS在快速检测绞碎牛肉中的大肠杆菌O157：H7方面的应用，以努力支持SESI-MS病原体挥发性数据库的建立企业和利基市场。RO1的建议可能集中在ENaC小鼠模型中PA非粘液-粘液转变的代谢特征。 2009/10年度接受/出版的出版物 1.Low S.Y.，Hill J.E.，and Peccia P.2009 DNA适配子识别烟曲霉的Asp f1变应原生化和生物物理研究通讯386(3)544-548 2.Johnson，N.R.和Hill，J.E.通过酶解家禽粪便改良土壤的磷组成：高通量方法的演示和对酶不稳定的P SSSAJ的提示 2009/10年度出版物正在审查中 1.Giles，C.D.，Cade-Menun，B.和Hill，J.E.(正在审查中)随时间跟踪的家禽粪便修正土壤中的磷迁移和转化 2.Giles，C.D.，Cade-Menun，B.和Hill，J.E.(审查中)土壤和粪便中的肌醇磷酸盐：丰度、循环和测量 3.Haznedaroglu B.Z.，Zorlu O.，Hill J.E.，Walker S.L.(正在审查中)确定鞭毛在运动性和非运动性肠炎沙门氏菌血清型运输中的作用 4.朱军，Bean H.，郭勇，Hill J.E.用SESI-MS快速检测细菌培养物中的顶空挥发性有机化合物