title: A biochemical fluorometric method for assessing the oxidative properties of HDL.
abstract: Most current assays of HDL functional properties are cell-based. We have developed a fluorometric biochemical assay based on the oxidation of dihydrorhodamine 123 (DHR) by HDL. This cell-free assay assesses the intrinsic ability of HDL to be oxidized by measuring increasing fluorescence due to DHR oxidation over time. The assay distinguishes the oxidative potential of HDL taken from different persons, and the results are reproducible. Direct comparison of this measurement correlated well with results obtained using a validated cell-based assay (r(2) = 0.62, P < 0.001). The assay can be scaled from a 96-well format to a 384-well format and, therefore, is suitable for high-throughput implementation. This new fluorometric method offers an inexpensive, accurate, and rapid means for determining the oxidative properties of HDL that is applicable to large-scale clinical studies.
authors.affiliation: Department of Medicine, University of California, Los Angeles, CA 90095. Electronic address: tkelesidis@mednet.ucla.edu.
authors.forename: Theodoros
authors.initials: T
authors.lastname: Kelesidis
authors.affiliation: Department of Medicine, University of California, Los Angeles, CA 90095.
authors.forename: Judith S
authors.initials: JS
authors.lastname: Currier
authors.affiliation: Department of Medicine, University of California, Los Angeles, CA 90095.
authors.forename: Diana
authors.initials: D
authors.lastname: Huynh
authors.affiliation: Department of Obstetrics and Gynecology, University of California, Los Angeles, CA 90095.
authors.forename: David
authors.initials: D
authors.lastname: Meriwether
authors.affiliation: Department of Medicine, University of California, Los Angeles, CA 90095.
authors.forename: Christina
authors.initials: C
authors.lastname: Charles-Schoeman
authors.affiliation: Department of Medicine, University of California, Los Angeles, CA 90095; Department of Obstetrics and Gynecology, University of California, Los Angeles, CA 90095; Department of Molecular and Medical Pharmacology, University of California, Los Angeles, CA 90095 and.
authors.forename: Srinivasa T
authors.initials: ST
authors.lastname: Reddy
authors.affiliation: Department of Medicine, University of California, Los Angeles, CA 90095.
authors.forename: Alan M
authors.initials: AM
authors.lastname: Fogelman
authors.affiliation: Department of Medicine, University of California, Los Angeles, CA 90095.
authors.forename: Mohamad
authors.initials: M
authors.lastname: Navab
authors.affiliation: Department of Medicine, University of California, Los Angeles, CA 90095; Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine, University of California, Los Angeles, CA 90095.
authors.forename: Otto O
authors.initials: OO
authors.lastname: Yang
mesh_terms: D000818:Animals; D016632:Apolipoprotein A-I / Q000494:pharmacology; D016477:Artifacts; D002474:Cell-Free System; D004305:Dose-Response Relationship, Drug; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism; D005470:Fluorometry / Q000379:methods*; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D008075:Lipoproteins, HDL / Q000378:metabolism*; D008297:Male; D051379:Mice; D010084:Oxidation-Reduction / Q000187:drug effects; D015203:Reproducibility of Results; D020112:Rhodamine 123 / Q000378:metabolism; D012235:Rhodamines / Q000378:metabolism; D013997:Time Factors
chemical_list: D016632:Apolipoprotein A-I; D005456:Fluorescent Dyes; D008075:Lipoproteins, HDL; D012235:Rhodamines; C058319:dihydrorhodamine 123; D020112:Rhodamine 123