The biochemistry core of this program project grant supports all the individual sections by providing sophisticated quantitative and qualitative assays for a number of lipids using mass spectrometric techniques. Most of the quantitation of lipids is carried out by stable isotope dilution mass spectrometry using isotope labeled internal standards to correct for losses during isolation and purification. Quantitative assays employ LC/MS, tandem mass spectrometry (LC/MS/MS), positive ion electron ionization GC/MS, and negative ion chemical ionization GC/MS as the major analytical techniques. Specific assays have been developed over the years and are a part of the routine assays available in the biochemistry core including quantitation of primary prostaglandins, leukotrienes, combinations of these arachidonate metabolites (eicosanoid profile), platelet activating factor (PAF), lyso-PAF, cholesterol, and the major phospholipid classes including glycerophosphocholine and glycerophosphoserine molecular species. In addition, fatty acids are quantitated as a fatty acid profile where the major fatty acid between Cn-C22 are analyzed in a single GC/MS run. Nonenzymatic oxidized metabolites of arachidonic acid (isoprostanes) are also quantitated by either GC/MS or LC/MS/MS techniques. Qualitative analyses are also carried out in the core facility as well as methods development when the specific need arises. In general, these techniques will employ mass spectrometry as the quantitative tool for precise and accurate assays. Methods development may also lead to the implementation of methods in the investigators own laboratory. The staff of the biochemistry core are skilled in the operation of mass spectrometers and the application of mass spectrometry in the development of precise methods used in lipid biochemistry.
| Zemski Berry, Karin A; Murphy, Robert C; Kosmider, Beata et al. (2017) Lipidomic characterization and localization of phospholipids in the human lung. J Lipid Res 58:926-933 |
| Mould, Kara J; Barthel, Lea; Mohning, Michael P et al. (2017) Cell Origin Dictates Programming of Resident versus Recruited Macrophages during Acute Lung Injury. Am J Respir Cell Mol Biol 57:294-306 |
| Gibbings, Sophie L; Thomas, Stacey M; Atif, Shaikh M et al. (2017) Three Unique Interstitial Macrophages in the Murine Lung at Steady State. Am J Respir Cell Mol Biol 57:66-76 |
| Frasch, S Courtney; McNamee, Eóin N; Kominsky, Douglas et al. (2016) G2A Signaling Dampens Colitic Inflammation via Production of IFN-?. J Immunol 197:1425-34 |
| Janssen, William J; Bratton, Donna L; Jakubzick, Claudia V et al. (2016) Myeloid Cell Turnover and Clearance. Microbiol Spectr 4: |
| Yun, Bogeon; Lee, HeeJung; Jayaraja, Sabarirajan et al. (2016) Prostaglandins from Cytosolic Phospholipase A2?/Cyclooxygenase-1 Pathway and Mitogen-activated Protein Kinases Regulate Gene Expression in Candida albicans-infected Macrophages. J Biol Chem 291:7070-86 |
| Desch, A Nicole; Gibbings, Sophie L; Goyal, Rajni et al. (2016) Flow Cytometric Analysis of Mononuclear Phagocytes in Nondiseased Human Lung and Lung-Draining Lymph Nodes. Am J Respir Crit Care Med 193:614-26 |
| Jayaraja, Sabarirajan; Dakhama, Azzeddine; Yun, Bogeon et al. (2016) Cytosolic phospholipase A2 contributes to innate immune defense against Candida albicans lung infection. BMC Immunol 17:27 |
| Kandasamy, Pitchaimani; Numata, Mari; Berry, Karin Zemski et al. (2016) Structural analogs of pulmonary surfactant phosphatidylglycerol inhibit toll-like receptor 2 and 4 signaling. J Lipid Res 57:993-1005 |
| Zemski Berry, Karin A; Murphy, Robert C (2016) Phospholipid Ozonation Products Activate the 5-Lipoxygenase Pathway in Macrophages. Chem Res Toxicol 29:1355-64 |
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