Abstract:

CWAF (COREXIT water-accommodated fraction) was fractionated using solid phase extraction using Silica cartridges. Four fractions were collected: 50:50 water:ethanol, methanol, dichloromethane and hexane. Fractions were then dried down and resuspended in DMSO (dimethyl sulfoxide) to be used for ligand binding assays. HEK293T cells were triple transfected with PPARγ (peroxisome proliferator-activated receptor gamma) ligand binding domain expression vectors, firefly luciferase reporter vectors and renilla luciferase control vectors. Cells were then exposed to positive and negative control ligands (Rosiglitazone and DMSO respectively) and serial dilutions of DOSS (dioctyl sodium sulfosuccinate), ICP (inductively coupled plasma):PG (propylene glycol):DOSS, ICP:PG and Span80 treatments. Activation of the PPARγ binding domain was observed in DOSS containing treatments only, exhibiting a dose response curve indicating obesogenic potential DOSS.

Suggested Citation:

John E. Baatz, Alexis Temkin, Demetri D. Spyropoulos. 2016. Effects of CWAF chemically enhanced water accommodated sub-fractions on peroxisome proliferator-activated receptor PPAR gamma binding.. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/N71R6NH5

Publications:

Temkin, A. M., Bowers, R. R., Magaletta, M. E., Holshouser, S., Maggi, A., Ciana, P., … Spyropoulos, D. D. (2015). Effects of Crude Oil/Dispersant Mixture and Dispersant Components on PPARγ Activity in Vitro and in Vivo: Identification of Dioctyl Sodium Sulfosuccinate (DOSS; CAS #577-11-7) as a Probable Obesogen. Environmental Health Perspectives, 124(1). doi:10.1289/ehp.1409672

Purpose:

Study the effects of oil fractions enhanced with DOSS surfactant on PPARG. PPARG regulates fatty acid storage and glucose metabolism. PPAR-gamma is involved in the pathology of numerous diseases including obesity, diabetes, atherosclerosis, and cancer.

Data Parameters and Units:

PPARg_Ligand Binding Assay CWAF.csv -- CWAF mixture was fractionated using solid phase extraction and four solvents, 50:50 water:ethanol, methanol, dichloromethane, and hexane. [NTC No Template Control, DMSO Dimethyl Sulfoxide, CWAF Chemically enhanced Water Accommodated Fractions 1:400, 10nM Rosi, 50/50 water ethanol 1:200, 50/50 water ethanol 1:100, methynol 1:200, methynol 1:100, DCM dichloromethane 1:200, DCM dichloromethane 1:100, hexane 1:200, hexane 1:100] verses [ratio 1, ratio 2, ratio 3, average, standard deviation]. PPARg_Ligand Binding Assay_CWAF Franctions.pdf -- bar plot of the csv file. PPARg_Ligand Binding Assay CWAF.xlsx -- raw and normalized of the csv file. PPARγ ligand binding domain expression vectors, firefly luciferase reporter vectors and renilla luciferase control vectors. PPARg_Ligand Binding Assay_CWAF Fractions_DIF.docx -- Fractionation Protocol (same as metadata abstract).

Individual Files: